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Objective: To explore the relationship between blood glucose variability, collateral circulation and basilar artery computed scan angiography score (Batman) and prognosis of mechanical thrombectomy with Solitaire stent in patients with large vascular occlusive APCI. Methods: A retrospective study was conducted on 113 patients with large vessel occlusive APCI who underwent Solitaire stent mechanical thrombectomy in the Department of Neurology of Nanyang Central Hospital from March 2021 to July 2022. According to the prognosis, they were divided into outcome group (46 cases) and adverse group (67 cases). Evaluate the prognosis based on the Modified Rankin Scale three months after the surgery. The differences in collateral circulation, GV and Batman score between the two groups were compared, and the related factors affecting the prognosis of large vessel occlusive APCI patients treated with Solitaire stent mechanical thrombectomy were analyzed by multivariate logistic regression model. Results: The age of 113 patients with acute large vessel occlusive APCI was (65.3±8.9) years old. The proportion of female was 34.5% (39 cases). Compared with the outcome group, the adverse group had a lower proportion of collateral circulation [40 cases (87.0%) vs 47 cases (70.2%)], higher GV score [(25.19±3.54) vs (30.36±4.11) points], lower Batman score [(7.49±1.52) vs (6.65±1.33) points], higher proportion of atrial fibrillation history [16 cases (23.9%) vs 4 cases (8.7%)], higher National Institutes of Health Stroke Scale (NIHSS) score at admission [(8.33±0.74) vs (7.25±0.92) points], larger core infarct volume [(32.57±4.87) vs (29.54±5.14) ml], and longer time from admission to vascular recanalization [(123.52±31.17) vs (102.47±29.54) min] (all P <0.05). Atrial fibrillation history, core infarct volume, NIHSS score at admission, time from admission to vascular recanalization, glycemic variability, collateral circulation, and Batman score were related factors for the prognosis of large vessel occlusive APCI patients treated with Solitaire stent mechanical thrombectomy, with OR values (95% CI ) of 1.383 (1.124-1.641), 1.166 (1.007-1.350), 4.777 (1.856-12.297), 3.068 (2.379-3.757), 1.477 (1.209-1.806), 0.742 (0.654-0.831), and 0.717 (0.214-1.221), respectively (all P <0.05). Conclusion: Blood glucose variation is a risk factor for prognosis of mechanical thrombectomy with Solitaire stent in patients with large vascular occlusive APCI, and collateral circulation and Batman score are protective factors.

Groundwater pollution risk assessment is an important part of groundwater pollution prevention and control. Taking groundwater in the plain area of Barkol-Yiwu Basin as the research object, the DRSTIW model was constructed to evaluate groundwater vulnerability. According to the survey data of contaminated sites and land use types, point source and non-point source pollutions were classified, and the groundwater pollution load was evaluated. The primary value, economic value, and ecological value of groundwater were considered to evaluate the functional value of groundwater. The groundwater pollution risk assessment map was generated by using the map algebra function of ArcGIS software. The ROC curve was used to verify the risk assessment results of groundwater pollution. The spatial distribution of cold and hot spots of groundwater pollution risk was obtained by calculating the G index, and the change in hot spots was quantitatively analyzed by combining the center of gravity and standard deviation ellipse. The results showed that the groundwater vulnerability was generally low. The gentle terrain slope, shallow groundwater depth, and strong aquifer permeability made it easy for pollutants to enter the aquifer, resulting in high groundwater vulnerability in the northwest of Santanghu Town, the southeast of Dahongliuxia Township, and Kuisu Town in Barkol County. The groundwater pollution load and groundwater functional value were generally low, whereas the point source and non-point source pollution caused by industrial and agricultural production and life would increase the groundwater pollution load in local areas, and the human economic activities and habitat quality levels would affect the distribution of high groundwater function value areas. Groundwater pollution risk was generally low; very high and high pollution risk areas only accounted for 6.8% of the study area, mainly distributed in Naomaohu Town and Yanchi Town of Yiwu County, Kuisu Town, Shirenzi Township, Huayuan Township of Barkol County, and Hongshan Farm of Corps. The shallow groundwater depth, strong permeability of soil surface and vadose zone media, poor adsorption capacity, and concentrated distribution of pollution sources made it easy for pollutants to migrate and enrich in these areas. Under the dual influence of high vulnerability and high pollution load of groundwater, the risk of groundwater pollution in local areas increased. Affected by human activities, there was a certain spatial agglomeration of groundwater pollution risk, and the overall trend was from northwest to southeast. The results of groundwater pollution risk assessment provided a scientific reference for the division of groundwater pollution prevention and control areas.

Groundwater contamination risk assessment is an effective tool for groundwater pollution prevention and control. The evaluation system mainly includes three parts:groundwater contamination source load assessment, groundwater vulnerability assessment, and groundwater function value evaluation. Taking the plain area of southern Turpan Basin as an example, based on the survey data and land use data, point source pollution and non-point source pollution were divided to evaluate the load of groundwater pollution sources, the classical DRASTIC model was selected to evaluate the vulnerability of groundwater, and the functional value of groundwater was evaluated from the point of view of water quality and quantity. The three factors were weighted and superimposed via GIS platform to generate the risk zoning map of groundwater contamination. The results showed that the overall risk of groundwater contamination in the study area was low. The area of high-risk and relatively high-risk areas accounted for 15.5% of the total study area, which were mainly distributed in L1, L2, and L3 of the study area. L1 was mainly affected by high pollution source load and high groundwater vulnerability. L2 was mainly the result of the joint action of high groundwater function value and domestic non-point source pollution. Non-point source pollution dominated by agricultural activities and high functional value of groundwater were the main reasons for the high risk of groundwater pollution in the L3 area. The results of the groundwater contamination risk assessment serve as an important reference for decision-makers to delineate the prevention and control area of groundwater pollution.

Lacrimal gland injection of botulinum toxin type A inhibits the secretion of tears. As a new method to treat or alleviate the symptom of tears or epiphora, it has the characteristics of simple operation, definite curative effect, repeatable treatment and no irreversible complications. It provides an optional treatment scheme for many patients with refractory tears or epiphora. This article reviews the pharmacological characteristics of botulinum toxin type A, the mechanism of inhibiting tear secretion, the method and dose of lacrimal gland injection, indications and contraindications, clinical efficacy evaluation, complications, existing problems to be solved and prospects for reference.

Objective: To understand the epidemiological characteristics of COVID-19 epidemic in Ejina banner, Inner Mongolia, in October 2021 and provide evidence for the improvement of COVID-19 prevention and control. Methods: The information about the time, area and population distributions of COVID-19 cases in Ejina before November 13, 2021 and the gene sequencing result of the isolates were collected for a statistical descriptive analysis. Results: The first COVID-19 case in Ejina occurred on 7 October, 2021. A total of 164 COVID-19 cases were reported from October 19 to November 12. Most cases were distributed in 6 communities in Darahub (156 cases, 95.12%). The result of full gene sequencing of the isolates indicted that the pathogen was Delta variant (B.1.617.2). The male to female ratio of the cases was 1.3∶1. The age of cases ranged from 1 to 85 years, and the cases aged 20-59 years accounted for 78.66%. The main clinical symptoms were sore throat (91 cases, 91.92%), cough (49 cases, 49.49%) and fever (23 cases, 23.23%). Most cases were ordinary ones (81 cases, 49.39%) and mild ones (68 cases, 41.46%). The cases were mainly detected at the isolation points (84 cases, 51.22%) and through population based nucleic acid testing (62 cases, 37.80%). The basic reproduction number ( R 0 ) of COVID-19 was 5.3, the average incubation period was 3.9 days. The local government rapidly started Ⅳ level emergency response and conducted 10 rounds of nucleic acid tests. The transferring of travelers reduced the risk for the further spread of COVID-19 in Ejina. Conclusions: The epidemic of COVID-19 in Ejina characterized by strong transmission, short incubation period, herd susceptibility and case clustering. Delta variant (B.1.617.2) was the pathogen, which might be imported from Zeke port. Comprehensive prevention and control measures, such as closed-loop management and vaccination, should be continued. The successful transferring of the patients and travelers provided evidence for the effective and precise prevention and control of COVID-19 in a routine manner.

Acetic acid is a common inhibitor present in lignocellulosic hydrolysate. Development of acetic acid tolerant strains may improve the production of biofuels and bio-based chemicals using lignocellulosic biomass as raw materials. Current studies on stress tolerance of yeast Saccharomyces cerevisiae have mainly focused on transcription control, but the role of transfer RNA (tRNA) was rarely investigated. We found that some tRNA genes showed elevated transcription levels in a stress tolerant yeast strain. In this study, we further investigated the effects of overexpressing an arginine transfer RNA gene tR(ACG)D and a leucine transfer RNA gene tL(CAA)K on cell growth and ethanol production of S. cerevisiae BY4741 under acetic acid stress. The tL(CAA)K overexpression strain showed a better growth and a 29.41% higher ethanol productivity than that of the control strain. However, overexpression of tR(ACG)D showed negative influence on cell growth and ethanol production. Further studies revealed that the transcriptional levels of HAA1, MSN2, and MSN4, which encode transcription regulators related to stress tolerance, were up-regulated in tL(CAA)K overexpressed strain. This study provides an alternative strategy to develop robust yeast strains for cellulosic biorefinery, and also provides a basis for investigating how yeast stress tolerance is regulated by tRNA genes.

Osimertinib-induced interstitial lung disease (ILD) is an uncommon, but fatal pulmonary toxicity in some patients. We report a case of a 64-year-old male with stage IV adeno-non-small cell lung cancer (NSCLC) harboring an exon 19 deletion in the epidermal growth factor receptor (EGFR) treated with osimertinib 80 mg/d for first-line targeted therapy. On day 60 after initiating treatment of osimertinib, the patient developed ILD. Osimertinib was discontinued immediately and oral prednisone 60 mg/d was initiated, ILD improved within 13 d. After balancing the risk and benefit, osimertinib was restarted concurrently with prednisone. The patient showed neither disease progression nor a recurrence of ILD for more than 16 months. Based on our case and literature review, retreatment with osimertinib under steroid coverage could be considered as an effective treatment option after careful risk-benefit assessment for patients with EGFR-mutant NSCLC.
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Objective: To analyze the effects of different types of sodium-glucose cotransporter 2 inhibitors (SGLT2i) on 24-hour ambulatory blood pressure in patients with type 2 diabetes mellitus and hypertension. Method: In this meta-analysis, we searched for randomized controlled trials on the effect of SGLT2i on 24-hour ambulatory blood pressure in patients with type 2 diabetes and hypertension. Three databases, namely PubMed, Web of Science and Cochrane Library, were searched. The search was organized on the concept of 3 conceptual groups: the first group contained terms used to describe SGLT2i, the second group contained terms related to blood pressure, and the third group contained terms used to describe randomized controlled trials. The search time was from the establishment of the database to December 2020. The inclusion and exclusion criteria were formulated in accordance with the requirements of the Cochrane systematic review. According to whether the heterogeneity of the study was significant or not, a random effect model or a fixed effect model were used to conduct the analysis on the impact of different types of SGLT2i on 24-hour ambulatory blood pressure and day and night blood pressure in patients with type 2 diabetes and hypertension. Further subgroup analysis was performed to define potential factors, which might lead to clinical heterogeneity. Results: Seven clinical trials were finally included. The result of the meta-analysis showed that compared with placebo group, SGLT2i could reduce the 24-hour dynamic systolic blood pressure of patients with type 2 diabetes and hypertension by 4.36 mmHg (1 mmHg=0.133 kPa). Reduction was 4.59, 3.74, 5.06, and 3.64 mmHg by canagliflozin, dapagliflozin, empagliflozin, and ertugliflozin respectively; SGLT2i could reduce the 24-hour dynamic diastolic blood pressure of patients with type 2 diabetes and hypertension by 2.20 mmHg, and the reduction was 2.30, 1.22, 2.00, and 2.69 mmHg by canagliflozin, dapagliflozin, empagliflozin and ertugliflozin respectively. SGLT2i could reduce the daytime systolic blood pressure of patients with type 2 diabetes and hypertension by 5.25 mmHg, and reduction was 5.38, 4.87, 6.00, and 4.37 mmHg by canagliflozin, dapagliflozin, empagliflozin and ertugliflozin, respectively. Simultaneously, SGLT2i could reduce the diastolic blood pressure of patients with type 2 diabetes and hypertension by 2.62 mmHg, and the reduction was 2.56, 2.47, and 2.80 mmHg by canagliflozin, empagliflozin and ertugliflozin, respectively. SGLT2i could reduce the nighttime systolic blood pressure of patients with type 2 diabetes and hypertension by 3.62 mmHg, and the reduction was 2.09, 2.06, 3.92, and 2.45 mmHg by canagliflozin, dapagliflozin, empagliflozin and ertugliflozin, respectively. At the same time, SGLT2i could reduce the nighttime diastolic blood pressure of patients with type 2 diabetes and hypertension by 1.60 and 1.51 mmHg, the reduction was 1.53 and 2.58 mmHg by canagliflozin, empagliflozin and ertugliflozin, respectively. Conclusion: SGLT2i can reduce 24-hour ambulatory blood pressure in patients with type 2 diabetes and hypertension.

The development and implement of microbial chassis cells can provide excellent cell factories for diverse industrial applications, which help achieve the goal of environmental protection and sustainable bioeconomy. The synthetic biology strategy of Design-Build-Test-Learn (DBTL) plays a crucial role on rational and/or semi-rational construction or modification of chassis cells to achieve the goals of "Building to Understand" and "Building for Applications". In this review, we briefly comment on the technical development of the DBTL cycle and the research progress of a few model microorganisms. We mainly focuse on non-model bacterial cell factories with potential industrial applications, which possess unique physiological and biochemical characteristics, capabilities of utilizing one-carbon compounds or of producing platform compounds efficiently. We also propose strategies for the efficient and effective construction and application of synthetic microbial cell factories securely in the synthetic biology era, which are to discover and integrate the advantages of model and non-model industrial microorganisms, to develop and deploy intelligent automated equipment for cost-effective high-throughput screening and characterization of chassis cells as well as big-data platforms for storing, retrieving, analyzing, simulating, integrating, and visualizing omics datasets at both molecular and phenotypic levels, so that we can build both high-quality digital cell models and optimized chassis cells to guide the rational design and construction of microbial cell factories for diverse industrial applications.

Objective: To evaluate whether ultrafine particulates (UFPs) have direct deleterious effects on cardiac function through activating MAPK signaling., Methods: Langendorff-perfused Sprague-Dawley rat hearts were randomly divided into 2 groups ( n =10/each group). In control group, the rat hearts were perfused with Tyrode's buffer for 40 min; in UFPs-treated group, the hearts were perfused with UFPs at a concentration of 12.5 mg/L. Cardiac function was determined by measuring left ventricular developed pressure (LVDP), left ventricular peak rate of contraction and relaxation (±d p /d t max ) and coronary flow (CF). The levels of malondialdehyde (MDA), superoxide dismutase (SOD), total anti-oxidant capacity (TAOC) were detected in order to evaluate cardiac oxidative stress via the thiobarbituric acid assay, water soluble tetrazolium salt assay and colorimetry, respectively. The expressions of p-p38 MAPK, p-ERKs and p-JNKs in the myocardium were observed using immunohistochemical staining and Western blots., Results: No significant changes in cardiac function were detected before and after the perfusion in control group while UFPs perfused hearts showed a decline in cardiac function in a time-dependent manner (all P < 0.05). In UFPs-treated group, LVDP, +d p /d t max , -d p /d t max and CF were statistically reduced from (82.6±2.1) mmHg, (1 624±113) mmHg/s, (1 565±116) mmHg/s, (12.0±0.2) mL/min to (56.8±4.4) mmHg, (1 066±177) mmHg/s, (1 082±134) mmHg/s, (8.7±0.3) mL/min (all P < 0.05), respectively. Furthermore, The comparison between the two groups observed that UFPs perfusion caused a significant decrease in cardiac function at 30 and 40 min compared with the control group (all P < 0.05). At the end of the perfusion, the level of MDA was increased from (0.98±0.14) nmol/L to (1.95±0.18) nmol/L, while SOD and TAOC were reduced from (12.50±1.87) U/mL and (6.83±1.16) U/mL to (6.50 ±1.04) U/mL and (3.67±0.82) U/mL (all P < 0.001) in UFPs group, respectively. In coincidence with these changes, immunohistochemistry and Western blots results showed that the levels of p-p38 MAPK, p-ERKs and p-JNKs in the myocardium significantly increased in UFPs group as compared with control group (all P < 0.05)., Conclusion: The results of this study demonstrated that the short-term exposure of UFPs to the isolated rat hearts has direct and acute toxic effects on cardiac function, probably related to attenuation of anti-oxidative capacity and activation of MAPK signaling pathways.

Objective: To analyze the disease burden of liver cancer in China. Methods: Based on eight data sources, including the series of Chinese Cancer Registry Annual Report, three national death cause surveys in China, China Health Statistical Yearbook, China Death Cause Surveillance Datasets, GLOBOCAN, Cancer Incidence in Five Continents (CI5), WHO Mortality Database and the Global Burden of Disease (GBD), the information on incidence, mortality and disability-adjusted life year (DALY) of liver cancer, were extracted for the analysis on the past, current and future disease burden caused by liver cancer in China. Results: 1) Past situation: The long-term data from 1973 to 2012 reported by the CI5 showed that in urban populations in China (taking Shanghai as an example), the incidence rate of liver cancer in males and females decreased by 41.3 % and 36.3 % , respectively, and that in rural areas (taking Qidong as an example) decreased by 32.3 % and 12.2 % , respectively. The Chinese Cancer Registry Annual Reports showed that the national incidence and mortality rates of liver cancer decreased by 8.1 % and 12.8 % respectively from 2005 to 2015. The Joinpoint analysis based on the data from the China Health Statistics Yearbook also showed a declining trend: the average annual percentage change of liver cancer mortality in China from 2002 to 2017 was -3.0 % ( P <0.05), and that in rural areas was -3.1 % ( P <0.05). 2) Current status: GLOBOCAN estimates that the rates of incidence, mortality and prevalence of liver cancer in China in 2018 were 18.3 per 100 000, 17.1 per 100 000 and 10.8 per 100 000, respectively. According to the latest annual report, the incidence and mortality rates of liver cancer in cancer registration areas in 2015 were 17.6 per 100 000 and 15.3 per 100 000, respectively, and both increased with age. The mortality rate was similar to that reported in 2017 (16.7 per 100 000) by the China Death Cause Surveillance Datasets, and the male to female ratio of live cancer deaths was estimated as 3.1. The GBD 2017 reports that the DALYs caused by liver cancer in China reached 11 153.0 thousand in 2017 (accounting for 53.7 % of the global DALYs) and hepatitis B virus infection was always the leading cause. 3) Prediction: The GLOBOCAN 2018 predicts that, by 2040, the number of liver cancer cases and deaths in China would reach 591 000 and 572 000 (with an increase of 50.5 % and 54.9 % , respectively, compared with those in 2018), with a more significant increase in people over 70 years old. 4) Economic burden: According to the literature review of economic burden data on liver cancer, the direct medical expenditure per patient with liver cancer generally showed a rising trend. Conclusions: Multiple data sources indicate that the incidence and mortality rates of liver cancer in populations in China decreased in the past decades, indicating the effect of population interventions. However, the population-level disease burden are still substantial, and comprehensive intervention strategies need to be continually strengthened and optimized, especially the primary and secondary prevention.

Objective: To evaluate the safety and efficacy of islet transplantation for patients with advanced diabetes. Methods: Five cases of islet allotransplantation were performed on 4 adult recipients. The same blood type adult brain-dead pancreas donors were selected and the islets were prepared in GMP laboratory. The prepared islet suspension was slowly injected into the liver of the recipients within 30-60 minutes. The immunosuppressive regimen was a combination of basiliximab, tacrolimus and mycophenolate mofetil and TNF-alpha monoclonal antibody was used to reduce the post-transplant inflammatory response. Insulin was temporarily applied to control blood glucose after surgery, and the dosage of insulin was adjusted to decrease according to the blood glucose level until it was discontinued. Results: A total of 5 islet transplants were performed in 4 patients, including 1 patient who received the second islet transplantations. All operations were succeed and the blood glucose and portal pressure were stable during the operation. Exogenous insulin was continued to keep blood glucose level stable (4-12 mmol/L) after surgery. Four cases (including the one who received two islet transplantation) started to stop using insulin at the third to fourth week, and the insulin dosage of the other case was 74% lower than that before the operation, and no hypoglycemic reaction occurred in all patients after islet transplantation. The C-peptide level in 3 patients reached the normal range, and the level in one patient with type I diabetes (without insulin release) remained at 0.45-0.6 μg/L (0.15-0.2 nmol/L). In addition, one patient showed a rise in blood glucose again and continued to use insulin half a year after insulin discontinuation. Then, he was performed the second islet transplantation which showed good effect and stopped taking insulin in 10 days after surgery. There were 3 cases of liver puncture bleeding after opeation, of which 2 cases were treated with ultrasound radiofrequency ablation to stop bleeding, 1 case stopped spontaneously, and no other complications were found. Conclusions: Islet transplantation is effective in the treatment of advanced diabetes patients with small trauma and high safety, which is worthy of more promotion. Long-term efficacy and maintenance therapy still need further investigation.

Objective: To investigate the acceptance and attitude toward a novel fecal immunochemical test (FIT) in colorectal cancer screening among populations in China. Methods: From May 2018 to May 2019, 2 474 people aged 50-74 years were recruited from five provinces of China (Zhejiang, Anhui, Jiangsu, Hunan and Yunnan). The general demographic characteristics, acceptance of the new FIT technology and operational difficulties through the whole screening process were obtained through questionnaire survey. Multivariate logistic regression model was used to analyze the factors related to difficulties encountered in sampling stool, reading and uploading results. Results: The subjects were (60.0±6.4) years old, and female, high school of above educated, unemployed/retired/other, married and with medical insurance status of "new rural cooperative medical care (NRCMC)" accounted for 61.7% (1 526), 29.0%(718), 34.3% (849), 92.7% (2 293) and 31.3%(775), respectively. The population's acceptance of the FIT technology was 94.8%. In the process of FIT screening, the percentage of occurred difficulties in sampling stool, reading and uploading results were 33.1% (819), 46.4% (1 147) and 62.9% (1 557), respectively. The main difficulties were the uncertainty about whether the sampling operation was standard (28.0%), the inability to accurately judge the result displayed (32.5%) and the need for help without using a smartphone (44.2%). The results of multivariate logistic regression model analysis showed that people aged 65-74 years old and with medical insurance status of "NRCMC" were more likely to encounter difficulties in sampling, and those who were unemployed/retired/other and living with 3 or more family members were less likely to encounter difficulties in sampling. Those aged 65-74 years old, farmers or migrant workers, and those with "NRCMC" were more likely to encounter difficulties in readingresults, and those with 3 or more family members were less likely to encounter difficulties in reading result. Those with "NRCMC" were more likely to encounter difficulties in uploading results, and those with education level of high school or above, living with more than 3 family members were less likely to encounter difficulties in uploading results. Conclusion: The acceptance of the new FIT technology is relatively high among the subjects. Age, education level, occupation, number of family members living together and medical insurance status might be related to difficulties encountered in sampling stool, reading and uploading results, and it can be further strengthened in terms of the technology and characteristics of sub-populations.

The evaluation of female fertility at first diagnosis is an important premise and basis for determining the treatment scheme of assisted reproduction. In this study, a survey was conducted among 13 assisted reproductive institutions in China to understand the evaluation indicators and detection methods of female fertility at first diagnosis in various institutions, and provide a basis for reasonable selection of indicators. The survey showed that the indicators of female fertility evaluation at first diagnosis among assisted reproductive institutions included general health indicators, ovarian reserve indicators, and uterine conditions, etc. The selection of indicators was considerably consistent, but the detection methods were quite different. Therefore, it is necessary to choose the detection method with better validity and less harm.

As a way of female fertility preservation, oocyte cryopreservation technology(OCT) has attracted more attention from the society. The health technology assessment (HTA) research progresses on OCT are important evidence basis for OCT clinical application promotion. Literatures on the maternal and offspring safety, efficacy and social ethics assessment of OCT were reviewed in this paper. Based on the current OCT evaluation evidence, short-term safety and effectiveness were confirmed, but long-term maternal and child safety should be testified in a large scale follow-up study. Social ethics evaluation methods of human assisted reproductive technology(ART) research were still in the exploratory stage. Therefore, it is necessary to establish the social ethics evaluation methods and system of human ART, including OCT, in China.

Objective: This study was to systematically update the economic evaluation evidence of colorectal cancer screening in mainland China. Methods: Based on a systematic review published in 2015, we expanded the scope of retrieval database (PubMed, EMbase, The Cochrane Library, Web of Science, CNKI, Wanfang Data, VIP, CBM) and extended it to December 2018. Focusing on the evidence for nearly 10 years (2009-2018), basic characteristics and main results were extracted. Costs were discounted to 2017 using the consumer price index of medical and health care being provided to the residents, and the ratio of incremental cost-effectiveness ratio (ICER) to per capita GDP in corresponding years were calculated. Results: A total of 12 articles (8 new ones) were included, of which 9 were population-based (all cross-sectional studies) and 3 were model-based. Most of the initial screening age was 40 years (7 articles), and most of the frequency was once in a lifetime (11 articles). Technologies used for primary screening included: questionnaire assessment, immunological fecal occult blood test (iFOBT) and endoscopy. The most commonly used indicator was the cost per colorectal cancer detected, and the median (range) of the 20 screening schemes was 52 307 Chinese Yuan (12 967-3 769 801, n =20). The cost per adenoma detected was 9 220 Yuan (1 859-40 535, n =10). In 3 articles, the cost per life year saved (compared with noscreening) was mentioned and the ratio of ICER to GDP was 0.673 (-0.013-2.459, n =11), which was considered by WHO as "very cost-effective" ; The range of ratios overlapped greatly among different technologies and screening frequencies, but the initial age for screening seemed more cost-effective at the age of 50 years (0.002, -0.013-0.015, n =3), than at the 40 year-olds (0.781, 0.321-2.459, n =8). Conclusions: Results from the population-based studies showed that the cost per adenoma detected was only 1/6 of the cost per colorectal cancer detected, and limited ICER evidence suggested that screening for colorectal cancer was generally cost-effective in Chinese population. Despite the inconclusiveness of the optimal screening technology, the findings suggested that the initial screening might be more cost-effective at older age. No high-level evidence such as randomized controlled trial evaluation was found.

Objective: The existed economic evaluations of cancer screening in Chinese population are almost all single-cancer focused, evidence on parallel comparison among multiple cancers is lacking. Thus, the aim of this study was, from a priority setting perspective, to compare the cost-effectiveness of six common cancers(colorectal cancer, breast cancer, liver cancer, lung cancer, esophageal cancer and stomach cancer) to facilitate policy making in future scaled-up screening in populations in China. Methods: Partially based on our previous single-cancer systematic reviews (colorectal cancer, breast cancer, liver cancer, and lung cancer), evidence of economic evaluations of cancer screening in populations in mainland China were systematically updated and integrated. The main updates include: 1) Stomach cancer and esophageal cancer were newly added to the current analysis. 2) The literature searching was extended to 8 literature databases, including PubMed, EMbase, The Cochrane Library, Web of Science, CBM, CNKI, Wanfang Data, and VIP. 3) The period of publication year was updated to the recent 10 years: January 1, 2009 to December 31, 2018. 4) The study focused on populations in mainland China. Following the standard processes of literature searching, inclusion and exclusion from previous systematic reviews, the basic characteristics, evaluation indicators and main results of the included studies were extracted. All the costs were discounted to 2017 value using the by-year consumer price index of medical and health care residents in China and presented in the Chinese Yuan (CNY). The ratios of incremental cost-effectiveness ratio (ICER) to China's per capita GDP in 2017 were calculated (<1 means very cost-effective, 1-3 means cost-effective, >3 means not cost-effective). Given a specific indicator, the median value among all reported screening strategies for each cancer was calculated, based on which priority ranking was then conducted among all cancers when data available. Results: A total of 45 studies were included, 22 for breast cancer, 12 for colorectal cancer, 6 for stomach cancer, 4 for esophageal cancer (all conducted in high-risk areas), 1 for liver cancer and none for lung cancer (was not then considered for next ranking due to limited numbers of studies). When based on the indicator, the median ratio of cost per life-year saved to China's per capita GDP (reported in 12 studies), the lowest ratio (-0.015) was observed in esophageal cancer among 16 strategies of 2 studies ( N =2, n= 16), followed by 0.297 for colorectal cancer ( N =3, n= 12), 0.356 for stomach cancer ( N =1, n= 4) and 0.896 for breast cancer ( N =6, n= 52, P (75)=3.602). When based on another commonly used ICER indicator, the median ratio of cost per quality-adjusted life-year gained to China's per capita GDP (reported in 13 studies), the least cost was found in stomach cancer (0.495, N =3, n= 8, P (75)=3.126), followed by esophageal cancer (0.960, N =1, n= 4, P (75)=1.762) and breast cancer (2.056, N =9, n= 64, P (75)=4.217). Data was not found for colorectal cancer. In addition, cost per cancer case detected was the most adopted indicator (32 studies). The median cost among all screening strategies for each cancer was 14 759 CNY for stomach cancer ( N =5, n= 7), 49 680 CNY for colorectal cancer ( N =12, n= 25) and 171 930 CNY for breast cancer ( N =13, n= 24), respectively. Data was not available for esophageal cancer and rare for precancer cases detected. Evidence related to cost per disability-adjusted life-year gained was not available. Conclusions: At China's national level and limited to the six cancers covered by the current study, the preliminary analysis suggests that stomach cancer and colorectal cancer were the most cost-effective target cancers and could be given priority in the future scaled-up screening in general populations. Esophageal cancer screening should be prioritized in high-risk areas. Breast cancer was also cost-effective in general but some of the intensive screening strategies were marginal. Data on liver cancer and lung cancer were too limited to conclude, and more well-designed studies and high-quality research evidence should be required. This priority ranking might be changed if other common cancers were involved analyses.

Objective: To study the safety of topical berberine solution in rabbit eyes and its effect on corneal epithelial repair in rabbit eyes. Methods: Experimental Study. Ninety-two Japanese rabbits were randomly divided into two groups by random number table method: the general group (32 rabbits, 64 eyes) and the corneal injury group (60 rabbits, 60 eyes).The general groups were further divided into 4 groups by random number table method, and each group has 8 rabbits (16 eyes). According to the administration of deionized water or 0.5, 1.0, 1.5 mg/ml berberine solution, they were divided into the general control group and the general A, B, and C group. Dosing with both eyes, each eye was given a single dose, and then it was given multiple times for 4 weeks after observation for 72h. After the corneal epithelium injury model made in the right eye of rabbits in the corneal injury groups, they were divided into a corneal injury control group and a corneal injury group A, B, and C according to the administration of deionized water or 0.5, 1.0, 1.5 mg/ml berberine solution. there were 5 rabbits (15 eyes) in each group, and the solutions were given continuously for 1 week. The rabbits in the general group were observed their behavioral changes, ocular surface and iris were scored by Draize eye irritation test scoring system. IOP was measured at different time points. Electroretinogram (ERG) was used to detect b-wave amplitude. In the corneal injury group, corneal epithelial defect repairment was observed at 1, 2, 3, 4, 5, 6, and 7 days after the corneal defect. Corneal histopathology observation after discontinuation of all rabbits. The pH value of rabbit tears was described by the paired t test, and the score of Draize eye irritation test were described by the rank-sum test. The analysis of variance and SNK- q were used for IOP, electroretinogram b-wave amplitude, corneal epithelial injury area and repair time. Results: No abnormal behavior was observed in the general group rabbits after single and multiple administration. There was no significant difference in the Draize eye irritation score among the general control group and the general group A, B, C at 1, 2, and 4 weeks of multiple administrations. Among them, the Draize eye irritation score of the general group C was 7 (0, 12), 6 (0, 10), 6 (0, 16) points (χ(2)=1.640, 0.265, 1.963, 1.381; P> 0.05).There were no significant difference in IOP at different times among the general control group and the general group A, B, C at different times ( F= 0.065, 0.292, 0.015, 0.041; P> 0.05). Before multiple administrations and after administration at 2, 4 weeks, the b-wave amplitudes of the general control group were (127.75±17.12), (129.18±15.83), (128.81±13.58) μV, and the general group A were (130.68).±18.75), (131.38±16.96), (130.62±12.18) μV,and the general group B were (128.00±16.74), (128.44±16.64), (129.06±16.16) μV, and the general group C were (131.81±19.37), (132.13±18.36), (129.94±12.60) μV. There was no statistically significant difference in b-wave amplitude in the groups at different times before and after administration ( F= 0.037, 0.011, 0.017, 0.702; P> 0.05). There was no significant difference in the results of corneal histopathology among the general control group and the general group A, B, C. The area of corneal epithelial defect in each corneal injury group was statistically significant at different time ( F= 5.316, 25.864, 127.613; P< 0.05). The corneal injury control group compared with the corneal injury group A, B, C, the corneal epithelial defect area in the corneal injury group C was significantly larger than the other three groups, with statistical differences ( q= 5.153, 10.313, 6.976; P< 0.05). The repair time of corneal epithelial in control group and the group A,B,C of corneal injury were (83.0±1.85), (82.9±2.07), (83.7±2.09) and (101.6±2.20) h. The corneal epithelium defect repair time in group C was longer and the difference was statistically significant ( F= 301.437, P= 0.000). Comparing the corneal injury control group and corneal injury group A and B, there was no statistical difference in the repair time of corneal epithelial defect ( F= 0.813, P= 0.450). After repair, there was no significant difference in the pathological results of the corneal tissue between the corneal injury groups. Conclusions: Berberine solution in rabbit eyes with topical application was safety, and has no obvious toxic effect on the ocular surface and ERG of normal rabbits. 1.5 mg/ml berberine solution delayed the repair of experimental corneal epithelial defect, but had no effect on the integrity of corneal tissue after repair. (Chin J Ophthalmol, 2020, 56: 131-137) .

Objective: To summarize clinical experience on the clinical feature, etiology and treatment of patients with spontaneous bloody tears as the initial symptom. Methods: Retrospective series of case studies. The clinical data and follow-up data of 27 cases of bloody tears as the first symptom in Lacrimal Center of Ophthalmology, the Third Medical Center of Chinese PLA General Hospital from June 2015 to December 2018 were reviewed. The clinical feature, specific cause, diagnosis, treatment and prognosis of these cases were evaluated. Results: A total of 27 cases were collected in this study. The patients were 10 males (37.0%) and 17 females (63.0%), including 21 adults (≥ 18 years old, 77.8%) and 6 minors (<18 years old, 22.2%). There were 22 monocular cases (81.5%) and 5 binocular cases (18.5%). Five cases (18.5%) were bleeding from the eye and other parts of the body, and 22 cases (81.5%) were bleeding only from the eye. There were 19 cases (70.4%) with hematic epiphora and secretions from the punctum, 3 cases (11.1%) with blood-stained tears, and 7 cases (25.9%) with blood-like tears. With regard to etiology, 6 cases (22.2%) were combined with systemic lesions, one of which was granulomatosis with polyangiitis and five of which (<18 years old) were idiopathic bloody tears. Twenty-one cases (77.8%) were local lesions, including 18 cases only involving the lacrimal system, 2 cases only involving the ocular surface, and 1 case involving both the lacrimal system and the ocular surface. Among the 21 cases with local lesions, 5 cases were induced by foreign body, 6 cases were induced by simple inflammation, and 10 cases were induced by tumor including 1 case with conjunctival benign tumor and 9 cases with tumor of the lacrimal system (5 with malignant tumor and 4 with benign tumor). Patients with idiopathic bloody tears received psychological and medical treatment, and interictal discharge was lengthened. One case of granulomatosis with polyangiitis was treated by trans-nasal endoscopic dacryocystorhinostomy. With the recurrence of granulomatosis and polyangiitis, bloody tears recurred after surgery. One patient with conjunctival hemangioma was untreated. Lesions in the lacrimal duct system were removed and dacryocystorhinostomy was performed. In this study, 2 patients (1 with small cell neuroendocrine carcinoma and 1 with adenoid cystic adenocarcinoma) died and the other had a good prognosis. Conclusions: Among the cases of bloody tears, adults and local lesions are more common. Most of the lesions are located in the lacrimal system and are tumors. The main treatment is to remove the lesions, and if necessary, to expand the resection and reconstruct the lacrimal duct. Idiopathic bloody tears occur in minors, who are gave psychotherapy and necessary medical treatment. (Chin J Ophthalmol, 2020, 56: 53 - 58) .

Objective: To understand the health literacy and relevant factors of cancer prevention consciousness in Chinese urban residents from 2015 to 2017. Methods: A cross-sectional survey was conducted in 16 provinces covered by the Cancer Screening Program in Urban China from 2015 to 2017. A total of 32 257 local residents aged ≥18 years old who could understand the investigation procedure were included in the study by using the cluster sampling method and convenient sampling method. All local residents were categorized into four groups, which contained 15 524 community residents, 8 016 cancer risk assessment/screening population, 2 289 cancer patients and 6 428 occupational population, respectively. The self-designed questionnaire was used to collect the information of demographic characteristics and cancer prevention consciousness focusing on nine common risk factors, including smoking, alcohol, fiber food, food in hot temperature or pickled food, chewing betel nut, helicobacter pylori, moldy food, hepatitis B infection, estrogen, and exercise. The logistic regression model was adopted to identify the influencing factors. Results: The overall health literacy of the cancer prevention consciousness was 77.4% (24 980 participants), with 77.4% (12 018 participants), 79.9% (6 406 participants), 77.2% (1 766 participants) and 74.5% (4 709 participants) in each group ( P< 0.001). The correct response rates for nine risk factors ranged from 55.2% to 93.0%. The multivariate logistic regression analysis showed that compared with community residents, people with primary school level education or below, and the number of people living together in the family <3, the cancer risk assessment/screening intervention population, cancer patients, those with junior high school level educationor above and the number of people living in the family ≥3 had better health literacy of the cancer prevention consciousness (all P values <0.05). Compared with females, 39 years old and below, government-affiliated institutions or civil servants, from the eastern region, males, older than 40 years, company or enterprise employees, and from the middle or western region had worse health literacy of the cancer prevention consciousness (all P values <0.05). Conclusion: The health literacy of the cancer prevention consciousness in Chinese urban residents should be improved. The cancer screening intervention, gender, age, education, occupation, the number of people co-living in the family, and residential region were associated with the health literacy of the cancer prevention consciousness.

Filamentous fungi are one of the platforms for producing fermented products. The specific characteristic of their submerged fermentation is the aggregation of mycelia that is affected by environmental conditions, leading to significantly different rheology for fermentation broth. Such a rheological change not only affects the transfer of mass, heat and momentum, but also the biosynthesis of target products and the efficiency of their production. In this article, strategies for morphological regulation of filamentous fungi are reviewed, and the impact of calcium signal transduction and chitin biosynthesis on apical growth of hyphae and branching of mycelia for their aggregation are further commented.

Objective: To analyze the impact of dual antiplatelet (DAPT) therapy combining with or without proton pump inhibitors (PPI) on the main outcomes after percutaneous coronary intervention (PCI). Methods: The PubMed, EMBASE and Cochrane Library were searched for relevant literature and the references obtained from these sources were retrieved manually from inception till September 2017. Inclusion and exclusion criteria were established follow the Cochrane review standard. A total of 977 literatures were included, 193 duplicates were excluded, 74 reviews, case reports, letters and systematic reviews were excluded, 667 literatures were excluded after reading the title and abstract, 34 literatures were excluded due to non-randomized control studies and unrelated outcome indicators, and 9 literatures were finally included with a total of 16 589 patients. RevMan 5.3 software was used to compare the incidence of major adverse cardiovascular events (MACE), cardiogenic death, recurrent myocardial infarction, target vessel revascularization, all-cause death, stent thrombosis, stroke, gastrointestinal bleeding and gastrointestinal events in patients with DAPT combining with or without PPI after PCI. Results: MACE was observed in 8 out of the 9 included literatures, and the results showed that MACE occurred in 561 out of 6 282 patients receiving DAPT combining with PPI therapy and in 951 out of 9 632 patients using DAPT alone ( OR= 1.15, 95 %CI 0.88-1.51, P> 0.05). Cardiogenic death was observed in 7 out of the 9 included literatures, and the results showed that cardiogenic death occurred in 172 out of 6 453 patients receiving DAPT combining with PPI treatment and in 321 out of the 9 839 patients using DAPT alone ( OR= 0.97, 95 %CI 0.80-1.18, P> 0.05). Recurrent myocardial infarction was observed in 7 out of the 9 included literatures, the results showed 416 out of 6 282 cases in DAPT combining with PPI therapy group experienced recurrent myocardial infarction and 691 out of 9 632 cases in DAPT group experienced recurrent myocardial infarction ( OR= 1.01, 95% CI 0.89-1.16, P> 0.05). Four out of 9 literatures observed revascularization. The results showed that revascularization was performed in 64 out of 2 173 patients receiving DAPT combining with PPI therapy and in 105 out of the 2 770 patients using DAPT alone ( OR= 1.33, 95 %CI 0.55-3.24, P> 0.05). All-cause death was observed in 7 out of the 9 included literatures, and the results showed that all-cause death occurred in 172 out of the 6 453 patients in DAPT combining with PPI therapy group and in 321 out of the 9 839 patients using DAPT alone ( OR= 0.97, 95 %CI 0.80-1.18, P> 0.05). Three out of the 9 included articles observed stent thrombosis, and the results showed that stent thrombosis occurred in 99 out of 2 997 patients receiving DAPT combining with PPI therapy and in 245 out of the 6 198 patients treated with DAPT ( OR= 1.07, 95 %CI 0.83-1.37, P> 0.05). Stroke was observed in 2 out of the 9 included literatures. The results showed that stroke occurred in 5 out of 2 019 patients receiving DAPT combining with PPI therapy, and in 4 out of the 2 033 patients treated with DAPT ( OR= 1.00, 95 %CI 0.29-3.49, P> 0.05). Gastrointestinal bleeding was observed in 6 out of the 9 included literatures. The results showed that gastrointestinal bleeding occurred in 26 out of 3 517 patients receiving DAPT combined with PPI therapy, and in 93 out of the 3 506 patients treated with DAPT, gastrointestinal bleeding was significantly lower in the DAPT combining with PPI group than DAPT alone group ( OR= 0.27, 95 %CI 0.17-0.41, P< 0.01). Gastrointestinal events were reported in 6 out of the 9 included articles. Similarly, gastrointestinal events were observed in 51 out of 3 517 patients receiving DAPT combined with PPI therapy, and in 190 out of the 3 506 patients treated with DAPT alone, the incidence of gastrointestinal events in the DAPT combined with PPI group was significantly lower than DAPT alone group ( OR= 0.24, 95 %CI 0.14-0.42, P< 0.01). Conclusions: The incidence of MACE, cardiogenic death, recurrent myocardial infarction, target vessel revascularization, all-cause death, stent thrombosis and stroke are not affected by DAPT combined with PPI therapy after PCI, while the incidence of gastrointestinal bleeding and gastrointestinal events could be reduced by adding PPI to DAPT in patients undergoing PCI.

Trichoderma reesei Rut-C30 is widely used in industrial cellulase production, and development of cellulase hyper-producer is of great importance for economic lignocellulosic biorefinery. In this study, T. reesei Rut-C30 was engineered with an artificial zinc finger proteins (AZFPs) library. Two mutants T. reesei M1 and M2 with improved cellulase production were obtained. Compared to the parent strain, the filter paper activity (FPase) of T. reesei M1 and M2 increased 100% and 53%, respectively. In addition, the total amount of extracellular protein from the M1 mutant increased 69%, whereas the endo-β-glucanase (CMCase) activity of the M2 mutant is 64% higher compared to the parental strain. Furthermore, RT-qPCR analysis showed that the major cellulase genes exhibited significantly increased expression in both mutants, but different patterns were observed in the two mutants. On the other hand, the cellulase transcriptional repressor ace1 was down-regulated in both mutants, but the transcription level of the activator xyr1 was only up-regulated in the strain M1. These results demonstrated that different AZFPs exert diverse regulatory mechanisms on cellulase production in T. reesei. Analysis of the target genes of AZFPs from T. reesei M1 and M2 will not only benefit further exploration of the regulatory mechanisms of cellulase biosynthesis in T. reesei, but also enable development of cellulase hyper-producing strains by metabolic engineering.

By-products released from pretreatment process of lignocellulose seriously hinder the development of cellulosic fuel ethanol. Therefore, the great way to increase the efficiency of cellulosic ethanol production is improvement of Saccharomyces cerevisiae tolerance to these inhibitors. In this work, the effects of LCB4 gene overexpression on cell growth and ethanol fermentation in S. cerevisiae S288C under acetic acid, furfural and vanillin stresses were studied. Compared to the control strain S288C-HO, the recombinant strain S288C-LCB4 grew better on YPD solid medium containing 10 g/L acetic acid, 1.5 g/L furfural and 1 g/L vanillin. Ethanol yields of recombinant strain S288C-LCB4 were 0.85 g/(L·h), 0.76 g/(L·h) and 1.12 g/(L·h) when 10 g/L acetic acid, 3 g/L furfural and 2 g/L vanillin were supplemented into the fermentation medium respectively, which increased by 34.9%, 85.4% and 330.8% than the control strain S288C-HO. Meanwhile, ethanol fermentation time was reduced by 30 h and 44 h under furfural and vanillin stresses respectively. Further metabolites analysis in fermentation broth showed that the recombinant strain produced more protective compounds, such as glycerol, trehalose and succinic acid, than the control strain, which could be the reason for enhancing strain tolerance to these inhibitors from pretreatment process of lignocellulose. The results indicated that overexpression of LCB4 gene could significantly improve ethanol fermentation in S. cerevisiae S288C under acetic acid, furfural and vanillin stresses.

Objective: To evaluate the clinical application and effect of preseting Double-J ureteric stent in percutaneous nephrolithotripsy. Method: 74 cases of renal calculi treated with PCNL in our hospital during June 2014 to February 2017 were retrospectively analyzed. Of 74 cases, 54 was male, 20 was female. All cases were aged 27 to 78, the mean age was (49.5±12.3) years old. The diameter of the stone was 20 to 59 mm, and the mean diameter was of (29.4±4.3) mm.Our Surgical methods was first putingFr6 double J tube in abnormal ureteral in advance in lithotomy position, then indwellingthree-way Foley catheter and clipping drainage port, perfusingirrigation port with 3 000 ml saline from 60-80 cm height.Perfusingsaline through irrigation port in prone position, we produce artificial hydronephrosis, then indwelling channel Fr20 through B ultrasound guided percutaneous nephrostomy, and removing renal calculi using holmium laser lithotripsy. Results: All patients were successfully completed percutaneous nephrostomy and indwell Fr20 channel, mean channel set up time (8.0 ±2.0) min, mean operation time (79±46) min, mean decline of hemoglobin (17.0±4.0) g/L, mean serum creatinine increased(3.1±1.1) μmol/L, one-stage stone-free rates 81.1%, complication rate 8.1% (1 case injured pleura and suffered from pneumothorax, 1 case suffered from massive hemorrhage of renal arteriovenous fistula after operation, 4 cases suffered postoperative fever). Conclusion: Advance in percutaneous nephrolithotripsy indwelling double J tube is a safe and feasible method, which is advantageous to the percutaneous renal puncture and the establishment of channels, and can avoid the blindness of along the line of indwelling double J.

Kluyveromyces marxianus, as unconventional yeast, attracts more and more attention in the biofuel fermentation. Although this sort of yeasts can ferment pentose sugars, the fermentation capacity differs largely. Xylose and arabinose fermentation by three K. marxianus strains (K. m 9009, K. m 1911 and K. m 1727) were compared at different temperatures. The results showed that the fermentation performance of the three strains had significant difference under different fermentation temperatures. Especially, the sugar consumption rate and alcohol yield of K. m 9009 and K. m 1727 at 40 ℃ were better than 30 ℃. This results fully reflect the fermentation advantages of K. marxianus yeast under high-temperature. On this basis, five genes (XR, XDH, XK, AR and LAD) coding key metabolic enzymes in three different yeasts were amplified by PCR, and the sequence were compared by Clustalx 2.1. The results showed that the amino acid sequences coding key enzymes have similarity of over 98% with the reference sequences reported in the literature. Furthermore, the difference of amino acid was not at the key site of its enzyme, so the differences between three stains were not caused by the gene level, but by transcribed or translation regulation level. By real-time PCR experiment, we determined the gene expression levels of four key enzymes (XR, XDH, XK and ADH) in the xylose metabolism pathway of K. m 1727 and K. m 1911 at different fermentation time points. The results showed that, for thermotolerant yeast K. m 1727, the low expression level of XDH and XK genes was the main factors leading to accumulation of xylitol. In addition, according to the pathway of Zygosaccharomyces bailii, which have been reported in NCBI and KEGG, the xylose and arabinose metabolic pathways of K. marxianus were identified, which laid foundation for further improving the pentose fermentation ability by metabolic engineering.

This study was designed to reverse multidrug resistance of lung cancer cells by downregulating MDR1 genes through RNA interference (RNAi) technology. A novel biodegradable cationic polymer (PEG- b- PLG-g-PEIs, GGI) was synthesized and characterized by 1H NMR. The particle size and zeta potential were measured by dynamic light scattering (DLS). The cell viability profile of GGI was tested by MTT method with both A549 and A549/DDP cell lines. Flow cytometry (FCM) technology was used to investigate the efficiency and intensity of delivering siRNA to cells by GGI polymer. RT-PCR and Western blot were used to detect the mRNA and P-gp expression after GGI/MDR1 siRNA transfection assay. The sensitivity of cisplatin administration after transfecting GGI/MDR1 siRNA polyplexs was performed with MTT and Annexin V-FITC/PI methods. The results suggest that the particle size and zeta potential of GGI/siRNA were 150 −200 nm and 16−28 mV. GGI exhibited a lower cell cytotoxity than PEI 25K and higher efficiency of delivering siRNA, which dramatically decreased the expression of MDR1 mRNA and P-gp of A549/DDP cells and increased much sensitivity to cisplatin in A549/DDP cells. GGI holds a great potential in gene delivery as a novel cationic polymer for further investigation.

Objective To investigate the role of Raf kinase inhibitor protein (RKIP) in the proliferation of LX-2 human hepatic stellate cells. Methods The recombinant plasmid siRNA-RKIP was transfected into LX-2 cells. Five days later, the stably transfected cells were screened and cultured. MTT assay was used to detect cell proliferation after RKIP was silenced. Cell apoptosis and cell cycle distribution were evaluated by flow cytometry. The expressions of α-smooth muscle actin (α-SMA) and collagen type 1 (Col1) mRNA were detected by quantitative real-time PCR. The expressions of RKIP, α-SMA, Col1 and extracellular signal-regulated kinases/mitogen-activated protein kinase (ERK/MAPK) signaling pathway related proteins were assessed by Western blot analysis. Results Compared with the control group, knockdown of RKIP significantly induced LX-2 cell proliferation, reduced cell apoptosis, raise cell number in G2, and increased the proteins and mRNA expressions of Col1 and α-SMA. Moreover, low-expression of RKIP significantly enhanced the phosphorylation of ERK/MAPK. Conclusion Knockdown of RKIP promotes LX-2 cell proliferation; its mechanism is related to the activation of ERK/MAPK signaling pathway.

Production of bioenergy and bio-based chemicals by using fermentable sugars released from low-cost renewable lignocellulosic biomass has received great attention. Efficient cellulolytic enzymes are crucial for lignocellulose bioconversion, but high cellulase production cost is limiting the bioconversion efficiency of cellulosic biomass and industrial applications of lignocellulose biorefinery. Studies on induction and regulation of cellulase in filamentous fungi will help to further develop superior fungal strains for efficient cellulase production and reduce cellulase production cost. With the advances in high-throughput sequencing and gene manipulation technology using fungal strains, an in-depth understanding of cellulase induction and regulation mechanisms of enzyme expression has been achieved. We reviewed recent progresses in the induction and regulation of cellulase expression in several model filamentous fungi, emphasizing sugar transporters, transcription factors and chromatin remodeling. Future prospects in application of artificial zinc finger proteins for cellulase induction and regulation in filamentous fungi were discussed.

To explore the effect of Mongolia Astragali Radix produced in Longxi of Gansu province in protecting cardiac and nephritic functions of patients of essential hypertension(EH) with metabolic syndrome(MetS). A total of two hundred and twenty-six EH patients with MetS aged above 18 were selected. Patients were randomly divided to control group(adopted conventional medical treatment), Astragali Radix group 1(added Astragali Radix capsules 10 g•d⁻¹ besides conventional medical treatment) and Astragali Radix group 2(added Astragali Radix capsules 5 g•d⁻¹ besides conventional medical treatment). Cardiac anatomy structure, cardiac systolic function and diastolic function were measured by M-mode echocardiography, two-dimensional echocardiography, Doppler echocardiographic determination and tissue Doppler imaging. The level of microalbuminuria(MAU) was evaluated by radioimmunoassay. In addition, the estimated glomerular filtration rate(eGFR) was calculated by modification of diet in renal disease (MDRD) formulas. The changes of relevant indicators for cardiac and nephritic functions before and after treatment were compared during the 12-month follow-up. The study protocol was registered at the website of Chinese clinical trial register and approved by the ethics committee of second hospital of Lanzhou university. Each patient was required to sign an informed consent. SPSS software was used for statistical analysis. According to the result, compare with before treatment, the three groups show no difference in efficacy of metablic indicators. Left ventricular end-systolic volume (ESV) and left ventricular end-systolic dimension (LVESd) of all patients were improved after treatment. However, there was no significant difference among the three groups. After the addition of Astragali Radix, the mitral flow velocity(Vp) of patients was improved to some extent(P<0.05). However, there was no significant difference among the three groups. Astragali Radix had a significant effect in reducing the MAU(P<0.05). Moreover, the MAU level of patients in Astragali Radix group 1 decreased more significantly than the other groups(P<0.05). Compared with conventional therapy, Astragali Radix combined with conventional therapy could improve cardiac structure, left ventricular systolic function, left ventricular diastolic function, and reduce the MAU to a certain extent in EH patients with MetS. Moreover, the effects of high-dose Astragali Radix are better than that of the low-dose Astragali Radix. However, the effect of Astragali Radix on EH patients with MetS shall be further observed to confirm its efficacy., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Objective: To explore the correlation between propagated sensation along meridian (PSM) and TCM constitution at different age stages., Methods: According to age, 840 participants were divided into a youth group (326 cases), a middle aged group (243 cases) and an elderly group (271 cases). The TCM constitution of all the participants was evaluated, and the PSM test was performed. The distribution of TCM constitution, the occurring rate and transmission of PSM in each group were observed and compared; the correlation between PSM and the TCM constitution was preliminary investigated by Logistic regression analysis., Results: The distribution of nine types of TCM constitution in three groups:the proportion of normal constitution and partial constitution were significantly different (all P <0.05); the occurring rate and transmission of PSM in three groups were not significantly different (all P >0.05); the proportion of occurring rate for nine types of TCM constitutions in the whole population, from high to low, presented special intrinsic quality, neutral quality, yin -deficiency quality, qi -deficiency quality, yang -deficiency quality, damp-heat quality, phlegm-dampness quality, qi -stagnation quality and blood-stasis quality; besides, the proportion of occurring rate for different TCM constitutions in the youth group, middle aged group and elderly group was similar to that of whole population. The Logistic regression analysis results indicated the neutral quality ( P =0.025) and special intrinsic quality ( P =0.018) were positively while blood-stasis quality ( P =0.043) was negatively related with PSM in all subjects; the qi -deficiency quality ( P =0.025), phlegm-dampness quality ( P =0.019), blood-stasis quality ( P =0.012) and qi -stagnation quality ( P =0.035) were negatively related with PSM in youth group; the neutral quality ( P =0.001) was positively related with PSM inthe middle aged group; the neutral quality ( P =0.006) and yin deficiency quality ( P =0.004) were positively related with PSM in the elderly group., Conclusions: The occurrence of PSM in different age stages is related with TCM constitution, which could be increased in clinical treatment to improve acupuncture efficacy.

Objective: To investigate whether tetrahydrobiopterin (BH 4 ) could improve left ventricular diastolic function through phosphoinositide-3 kinase/protein kinase B (PI 3 K/Akt) signaling pathway in hypertensive mice. Methods: Ten-week-old male C57BL/6 mice were used to establish the deoxycorticosterone acetate (DOCA)-salt hypertensive model, age matched Sham mice serve as the controls. Mice were divided into four groups: Sham( n =20), Sham+ BH 4 ( n =20), DOCA ( n =22), and DOCA+ BH 4 ( n =22). On the 14 days after surgery, mice in Sham+ BH 4 and DOCA+ BH 4 groups received BH 4 (0.1 ml/10 g) supplement for 7 days, while mice in Sham and DOCA groups were given equal volume of normal saline.The blood pressure measurements were performed 7 days later.Hemodynamic and echocardiographic parameters were used to assess left ventricular functions.High performance liquid chromatography (HPLC) analysis was used to measure cardiac biopterins BH 4 and BH 2 .The phosphorylated phospholamban (p-PLB) was detected by immunohistochemical staining. PI 3 K, Akt and phosphorylated Akt were assayed with Western blot analysis. Results: (1) The systolic and diastolic blood pressure of DOCA group were significantly higher than control group ( P <0.05). Compared with DOCA group, the systolic blood pressure was lower in DOCA+ BH 4 mice ( P =0.027). Diastolic blood pressure was similar between the groups. (2) Compared with Sham group, the left ventricular diastolic function indexes such as mitral annulus velocity (E') and E'/A'ratio were significantly lower, while the E/ E'ratio was significantly higher( P <0.05)in DOCA mice. The E/ E'ratio of DOCA+ BH 4 group was significantly lower than that of DOCA group ( P <0.05). Compared with Sham group, the left ventricular end-diastolic pressure (LVEDP), left ventricular end-diastolic pressure volumetric coefficient (EDPVR) and left ventricular relaxation time constant Tau index were significantly higher in DOCA mice ( P =0.002, 0.011 and 0.016, respectively). The EDPVR and Tau index were significantly lower in DOCA+ BH 4 group than in DOCA group ( P <0.05). (3) Compared with Sham group, the myocardial contents of BH 4 and BH 2 were significantly lower in DOCA mice ( P <0.05). The BH 4 levels and BH 4 /BH 2 ratio were significantly higher in Sham+ BH 4 and DOCA+ BH 4 groups than in the DOCA group ( P <0.05), but the BH 2 levels were similar between groups. (4) The cGMP content, SOD activity and NO content in the left ventricular myocardial tissue were significantly lower ( P <0.05), while the MDA content was significantly higher in DOCA mice than in Sham mice.The NO content and SOD activity in DOCA+ BH 4 groups were significantly higher than in the DOCA group ( P <0.05). (5) Compared with DOCA group, the expression of p-PLB was significantly higher in Sham mice and lower in DOCA+ BH 4 mice ( P <0.05). (6) The expression of PI 3 K, Akt and p-Akt (Ser473 and Thr308) in DOCA mice were significantly lower than in Sham group ( P <0.05). The expression of PI 3 K, Akt and p-Akt (Ser473) was significantly higher in DOCA+ BH 4 group than in DOCA group ( P <0.05). p-Akt (Thr308) expression was similar between DOCA + BH 4 group and DOCA group (all P >0.05). Conclusion: Our results suggest that BH 4 could improve left ventricular diastolic function in hypertensive mice, this effect might be mediated by reducing the oxidative stress in ventricular myocardium through modlating the expression of Akt and PLB phosphorylation.

Objective: To develop a method of high-resolution melting curve analysis (HRM) for investigating the association of single nucleotide polymorphisms (SNPs) of IL-1β and IL-1Ra genes with susceptibility to coronary heart disease (CHD) and serological indexes in CHD., Methods: PCR-HRM assays were established for detecting four SNPs (IL-1B-511C>T, IL-1B-31C>T, IL-1B+3954C>T and IL-1RNT>C) in 260 patients with CHD and 274 healthy controls. A case-control study was performed for analyzing the relationships of the four SNPs with CHD risk and serological indexes., Results: No significant statistical difference was observed between the four SNPs and the susceptibility to CHD, however, the result of gender stratification showed that there was the statistical differences in the male IL-1B-31C>T and the female IL-1B+3954C>T allele frequencies (P=0.039, 0.032, respectively). The result of haplotype analysis of the four SNPs showed that haplotype of IL-1B-511T/IL-1B-31C/IL-1B+3954C/IL-1RN T (TCCT) had a significant correlation with obviously increased CHD risk, but without statistical difference (OR=1.217, 95%CI: 0.950-1.559, P=0.123), while those of CTCC and TTCT had significant correlations with decreased CHD risk (OR=0.612, 95%CI: 0.376-0.994, P=0.046; OR=0.365, 95%CI: 0.154-0.862, P=0.017). No association was found between the four SNPs and the serological indexes in CHD subjects., Conclusion: The developed PCR-HRM detection assays based on HRM technology has been verified by direct sequencing and clinical specimen detection to be a rapid and homogeneous detection method for genotyping of the four SNPs. The SNPs of IL-1B-31C>T, IL-1B+3954C>T and the haplotypes of CTCC and TTCT are associated with the susceptibility to CHD in Chinese Han population of Lanzhou region.

Microalgae have been identified as promising candidates for biorefinery of value-added molecules. The valuable products from microalgae include polyunsaturated fatty acids and pigments, clean and sustainable energy (e.g. biodiesel). Nevertheless, high cost for microalgae biomass harvesting has restricted the industrial application of microalgae. Flocculation, compared with other microalgae harvesting methods, has distinguished itself as a promising method with low cost and easy operation. Here, we reviewed the methods of microalgae harvesting using flocculation, including chemical flocculation, physical flocculation and biological flocculation, and the progress and prospect in bio-flocculation are especially focused. Harvesting microalgae via bio-flocculation, especially using bio-flocculant and microalgal strains that is self-flocculated, is one of the eco-friendly, cost-effective and efficient microalgae harvesting methods.

Objective: To observe practicality and safety of CT-RIDC for the patients with presaccular lacrimal obstruction and study the related factors. To provide guidance for the diagnosis and treatment of the patients with presaccular lacrimal obstruction., Methods: Fifty-four patients (75 eyes) with presaccular lacrimal obstruction, including 3 cases (5 eyes) of upper and lower lacrimal punctum atresia, 15 cases (24 eyes) of superior and inferior canalicular obstruction, 18 cases (28 eyes) of common canaliculus obstruction, 18 cases (18 eyes) of old laceration of superior and inferior canalicular, were recruited. All patients underwent the examination by using 0° and 30° ear endoscope and the imaging characteristics of the inferior orifice of nasolacrimal duct were obtained. Fifty-three cases (65 eyes) with the opening inferior orifice were randomly divided into four groups (15 eyes, 17 eyes, 17 eyes, 16 eyes) according to different angle of head hypsokinesis in the supine position. The OM lines back along the sagittal plane of the head back 10°, 20°, 30°, 40° were measured. The angles of head hypsokinesis were set in accordance with the OM line measurement results.Intubation was successful when the intubation guided needle within the set of epidural anesthetic catheter aided with endoscopy was inserted into the inferior orifice of nasolacrimal duct and then the epidural anesthetic catheter was inserted into the lacrimal duct 6 mm. The angle (θ) between the long axis of the curved part of the intubation guided needle and the long axis of the guided needle was measured. The numbers of intubation success cases in each group were recorded.χ² test was used to compare intubation success rate under the different angle of head hypsokinesis.Scheffe method was used to compare intubation success rate between each group. Contrast medium was then injected into lacrimal duct through the epidural anesthetic catheter for patients with successful intubation and CT scan was carried out. 3D model of lacrimal passage was reconstructed with CT axial scanning., Results: The intubation guided needle was successfully inserted into the inferior orifice of nasolacrimal duct in 44 eyes among 65 eyes with the opening orifice. The number of successful intubation cases were 4 eyes, 15 eyes, 13 eyes, 12 eyes and the intubation success rate was 26.67%, 88.24%, 76.74%, 75.00% according the angle of head hypsokinesis 10° group (15 eyes), 20° group (17 eyes), 30° group (17 eyes), 40° group (16 eyes). The intubation success rate of head hypsokinesis 10° group was significantly lower than that of the other groups. Multiple Comparison by Scheffe Test demonstrated that there was a remarkable difference (P < 0.05) in the intubation success rate between head hypsokinesis 10° group and the other three groups. There were no significant differences in the intubation success rate among head hypsokinesis 20° group, head hypsokinesis 30° group and head hypsokinesis 40° group. The mean of θ was 108° with a range of 93.2°-120.5° by measuring the angle θ of the successful intubation cases. According to the frequency distribution plot, most θ was in 104°-115°. The successful intubation cases (44 eyes) underwent CT scan in the same position immediately after injected contrast medium (Optiray) through the epidural anesthetic catheter. CT images of 28 eyes showed retained contrast medium in the nasolacrimal duct and lacrimal sac. The findings from CT-DCG images were basically agreed with the operation., Conclusions: CD-RIDC may apply to pre-operative examination for the patients with the opening orifice and presaccular lacrimal obstruction.It provideed imaging evidence for diagnosis and therapy and it was safe and practical. Many factors could affect CD-RIDC. The main verified facors were the shape of the inferior orifice of nasolacrimal duct, head position of the patient and the bending angle of the tip of intubation guided needle.

Chromosomal integration enables stable phenotype and therefore has become an important strategy for breeding of industrial Saccharomyces cerevisiae strains. pAUR135 is a plasmid that enables recycling use of antibiotic selection marker, and once attached with designated homologous sequences, integration vector for stable expression can be constructed. Development of S. cerevisiae strains by metabolic engineering normally demands overexpression of multiple genes, and employing pAUR135 plasmid, it is possible to construct S. cerevisiae strains by combinational integration of multiple genes in multiple sites, which results in different ratios of expressions of these genes. Xylose utilization pathway was taken as an example, with three pAUR135-based plasmids carrying three xylose assimilation genes constructed in this study. The three genes were sequentially integrated on the chromosome of S. cerevisiae by combinational integration. Xylose utilization rate was improved 24.4%-35.5% in the combinational integration strain comparing with that of the control strain with all the three genes integrated in one location. Strain improvement achieved by combinational integration is a novel method to manipulate multiple genes for genetic engineering of S. cerevisiae, and the recombinant strains are free of foreign sequences and selection markers. In addition, stable phenotype can be maintained, which is important for breeding of industrial strains. Therefore, combinational integration employing pAUR135 is a novel method for metabolic engineering of industrial S. cerevisiae strains.

Zinc-finger proteins have been widely studied due to their highly conserved structures and DNA-binding specificity of zinc-finger domains. However, researches on the zinc-finger proteins from microorganisms, especially those from prokaryotes, are still very limited. This review focuses on the latest progress on microbial zinc-finger proteins, especially those from prokaryotes and the application of artificial zinc-finger proteins in the breeding of robust strains. Artificial zinc-finger proteins with transcriptional activation or repression domain can regulate the global gene transcription of microbial cells to acquire improved phenotypes, such as stress tolerance to heat, ethanol, butanol, and osmotic pressure. Using the zinc-finger domain as DNA scaffold in the construction of enzymatic system can enhance the catalytic efficiency and subsequently the production of specific metabolites. Currently, zinc-finger domains used in the construction of artificial transcription factor are usually isolated from mammalian cells. In the near future, novel transcription factors can be designed for strain development based on the natural zinc-finger domains from different microbes, which may be used to regulate the global gene expression of microbial cells more efficiently.

Industrial microorganisms are subject to various stress conditions, including products and substrates inhibitions. Therefore, improvement of stress tolerance is of great importance for industrial microbial production. Acetic acid is one of the major inhibitors in the cellulosic hydrolysates, which affects seriously on cell growth and metabolism of Saccharomyces cerevisiae. Studies on the molecular mechanisms underlying adaptive response and tolerance of acetic acid of S. cerevisiae benefit breeding of robust strains of industrial yeast for more efficient production. In recent years, more insights into the molecular mechanisms underlying acetic acid tolerance have been revealed through analysis of global gene expression and metabolomics analysis, as well as phenomics analysis by single gene deletion libraries. Novel genes related to response to acetic acid and improvement of acetic acid tolerance have been identified, and novel strains with improved acetic acid tolerance were constructed by modifying key genes. Metal ions including potassium and zinc play important roles in acetic acid tolerance in S. cerevisiae, and the effect of zinc was first discovered in our previous studies on flocculating yeast. Genes involved in cell wall remodeling, membrane transport, energy metabolism, amino acid biosynthesis and transport, as well as global transcription regulation were discussed. Exploration and modification of the molecular mechanisms of yeast acetic acid tolerance will be done further on levels such as post-translational modifications and synthetic biology and engineering; and the knowledge obtained will pave the way for breeding robust strains for more efficient bioconversion of cellulosic materials to produce biofuels and bio-based chemicals.

Propionic acid, a major inhibitor to yeast cells, was accumulated during continuous ethanol fermentation from corn meal hydrolysate by the flocculating yeast under stillage backset conditions. Based on its inhibition mechanism in yeast cells, strategies were developed for alleviating this effect. Firstly, high temperature processes such as medium sterilization generated more propionic acid, which should be avoided. Propionic acid was reduced significantly during ethanol fermentation without medium sterilization, and concentrations of biomass and ethanol increased by 59.3% and 7.4%, respectively. Secondly, the running time of stillage backset should be controlled so that propionic acid accumulated would be lower than its half inhibition concentration IC50 (40 mmol/L). Finally, because low pH augmented propionic acid inhibition in yeast cells, a higher pH of 5.5 was validated to be suitable for ethanol fermentation under the stillage backset condition.

Ethanol tolerance is related to the expression of multiple genes, and genome-based engineering approaches are much more efficient than manipulation of single genes. In this study, ultraviolet (UV) mutagenesis, dielectric barrier discharge (DBD) air plasma mutagenesis, and artificial transcription factor (ATF) technology were adopted to treat an industrial yeast strain S. cerevisiae Sc4126 to obtain mutants with improved ethanol tolerance. Mutants with high ethanol tolerance were obtained, and the ratio of positive mutants was compared. Among the three approaches, the rate of positive mutation obtained by ATF technology was 10- to 100-folds of that of the two other methods, with highest genetic stability, suggesting the ATF technology promising for rapid alteration of phenotypes of industry yeast strains for efficient ethanol fermentation.

Objective: To explore the relationship of alcoholism between osteoporosis or femoral head necrosis., Methods: In this case-control study, we selected 95 eligible patients with femoral head necrosis and another 67 cases of osteoporosis as case group, together with 342 patients of fractures from the Second Hospital affiliated to Shanxi Medical College, from February to December 2010, as the control group. Questionnaire was used to collect general information of the patients. Through comparative analysis, related factors of femoral head, osteoporosis were defined. 18 patients with alcoholic femoral head necrosis, 11 patients with alcoholic osteoporosis and 20 patients with fractures were selected from the above said three groups and going through the Alcohol Use Disorders Identification Test (AUDIT) as well as the Alcohol Use Disorders Scale(ADS). Using SPSS 13.0 conducted one-way ANOVA(analysis of variance), chi-square test, categorical logistic regression analysis were used for statistical analysis., Results: Results from logistic regression analysis showed that the adjusted odds ratio of those subjects who liked drinking alcohol had an incidence of femoral head necrosis or osteoporosis as 7.70 (95% CI:1.84, 32.30) and 8.44 (95% CI:1.70, 41.90), respectively. The risks of using hormone for treating femoral head necrosis or osteoporosis were 78.43 (95%CI:11.20, 149.05) and 22.75 (95%CI:2.59, 100.27) times than those without. Data from the AUDIT showed that:over-dose of alcohol drinking habit existed 100% in the femoral head necrosis group while 54.45% in the osteoporosis group, while 75 percent patients in the fractures group had normal alcohol drinking habit. Statistically significant differences appeared in the three groups (P < 0.01)., Results: from the ADS showed that there were statistically significant differences between the ADS scores of the three groups(F = 3.68, P = 0.03)., Conclusion: Alcohol intake did seem to be highly correlated with the incidence rates of femoral head necrosis or osteoporosis. Alcohol-related necrosis could be viewed as alcohol-dependent diseases while alcohol-related and osteoporosis could partially be recognized as alcohol-dependent disease.

Dye-decolorizing peroxidase (DyP-type peroxidase) represents a group of heme-containing peroxidases able to decolour various organic dyes, most of which are xenobiotics. To identify and characterize a new DyP-type peroxidase (ZmDyP) from Zymomonas mobilis ZM4 (ATCC 31821), ZmDyP was amplified from the genomic DNA of Z. mobilis by PCR, and cloned into the Escherichia coli expression vector pET-21b(+). Alignment of the amino acid sequence of ZmDyP with other members of the DyP-type peroxidases revealed the presence of the active site conserved residues D149, R239, T254, F256 as well as the typical GXXDG motif, indicating that ZmDyP is a new member of the Dyp-type peroxidase family. pET-21b(+) containing ZmDyP gene was expressed in E. coli by IPTG induction. The expressed enzyme was purified by Ni-Chelating chromatography. SDS-PAGE analysis of the purified enzyme revealed a molecular weight of 36 kDa, whereas activity staining gave a molecular weight of 108 kDa, suggesting that the enzyme could be a trimer. In addition, ZmDyP is a heme-containing enzyme as shown by a typical heme absorption peak of Soret band. Moreover, ZmDyP showed high catalytic efficiency with 2, 2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) as a substrate. These results enrich the pool of DyP-type peroxidases and lay a foundation for further studies.

Breeding of robust industrial Saccharomyces cerevisiae strains with high ethanol tolerance is of great significance for efficient fuel ethanol production. Zinc finger proteins play important roles in gene transcription and translation, and exerting control on the regulation of multiple genes. The sequence and localization of the zinc finger motif can be designed and engineered, and the artificial zinc finger protein can be used to regulate celluar metabolism. Stress tolerance of microbial strains is related to multiple genes. Therefore, it is possible to use artificially-designed zinc finger proteins to breed stress tolerant strains. In this study, a library containing artificial zinc finger protein encoding genes was transformed into the model yeast strain S288c. A recombinant strain named M01 with improved ethanol tolerance was obtained. The plasmid in M01 was isolated, and then transformed into the industrial yeast strain Sc4126. Ethanol tolerance of the recombinant strain of Sc4126 were significantly improved. When high gravity ethanol fermentation using 250 g/L glucose was performed, comparing with the wild-type strain, fermentation time of the recombinant strain was decreased by 24 h and the final ethanol concentration was enhanced by 6.3%. The results of this study demonstrate that artificial zinc finger proteins are able to exert control on stress tolerance of yeast strains, and these results provide basis to construct robust industrial yeast strains for efficient ethanol fermentation.

Consolidated bioprocessing technology can be used for Kluyveromyces marxianus YX01 to produce ethanol from Jerusalem artichoke, which is one of the potential processes to produce biofuel from non-cereal crops. In this study, we combined the aeration rate with the substrate concentration to conduct cross-over experiments for K. marxinaus YX01, and studied ethanol fermentation and the influence of inulin enzyme activity. The substrate concentration had a little repressive effect on ethanol productivity. When substrate concentration reached 250 g/L under anaerobic conditions, ethanol concentration was 84.8 g/L, and ethanol yield was reduced from 86.4% (50 g/L substrate concentration) to 84.7% of the theoretical value. Aeration rate could accelerate K. marxinaus YX01 ethanol fermentation, but reduced ethanol yield. When substrate concentration reached 250 g/L under aeration at 1.0 vvm, ethanol yield was reduced from 84.7% under anaerobic conditions to 73.3% of the theoretical value. With increased concentration of the carbon source and reduced aeration rate, the inulinase of K. marxinaus YX01 reduced and the concentration of glycerol increased, however, the acetic acid increased with the increased concentration of the carbon source and aeration rate. When substrate concentration reached 250 g/L under anaerobic conditions, inulinase activity was only 6.59 U/mL; when substrate concentration reached 50 g/L under aeration at 1.0 vvm, inulinase activity was 21.54 U/mL.

We used ribosome engineering technology, with which antibiotic-resistant strains are resulted from mutations on microbial ribosome, to screen a high butanol-producing Clostridium strain. A novel mutant strain S3 with high butanol production and tolerance was obtained from the original Clostridium acetobutylicum L7 with the presence of mutagen of streptomycin. Butanol of 12.48 g/L and ethanol of 1.70 g/L were achieved in S3, 11.2% and 50%, respectively higher than the parent strain. The conversion rate of glucose to butanol increased from 0.19 to 0.22, and fermentation time was 9 h shorter. This caused an increase in butanol productivity by 30.5%, reaching 0.24 g/(Lh). The mutant butanol tolerance was increased from 12 g/L to 14 g/L, the viscosity of fermentation broth was dramatically decreased to 4 mPa/s, 60% lower than the parent strain. In addition, the genetic stability of mutant strain S3 was also favorable. These results demonstrate that ribosome engineering technology may be a promising process for developing high butanol-producing strains.

Objective: To investigate immune state in lung of BALB/c mice with ovalbumin (OVA) allergy and the effects of fulvotomentoside (Ful) on lungs of the mice and provide some clues for the mechanism that patients with food allergies were prone to asthma and observe the effects of the treatment with traditional Chinese medicine., Method: Ninety-six female BALB/c mice were randomly divided into 6 groups. Mice in group 1 and group 2 were sensitized intraperitoneally and challenged intragastrically with OVA and were exposed to phosphate buffer solution and OVA respectively by nebulized inhalation. Mice in group 3 and group 4 were treated with Ful, other processes were the same as the mice in group 1 and group 2, respectively. Mice in group 5 were not challenged intragastrically with OVA and other processes were the same as the mice in group 2. Group 6 was the control group. The number of total leukocytes and cell classification in bronchoalveolar lavage (BALF) were counted, and inflammatory characteristic of lung was scored by staining with hematoxylin and eosin. The protein expressions of transforming growth factor (TGF-β1), interleukin-6 (IL-6), interleukin-17 (IL-17A) in lung of the mice were detected by immunohistochemical method. The activation of neutrophils in lung was assayed by the level of myeloroxidase (MPO)., Result: There was no inflammatory cells infiltration in lung of the mice in group 1. Compared with group 6, numbers of total leukocytes and erythrocytes as well as the percentage of neutrophils and lymphocytes were increased in group 2. Inflammatory score and protein expressions of TGF-β1 [(75 437 ± 3 638) vs. (6 118 ± 1 978)], IL-6 [(121 650 ± 25 389) vs. (15 726 ± 9 360)], IL-17A [(252 105 ± 31 651)vs. (72 644 ± 12 285)] in lung were increased, too. Inflammatory score and TGF-β1 (11 054 ± 1 468), IL-6 (50 877 ± 11 744), IL-17A (137 864 ± 28 986) expressions in group 5 were lower than those in group 2. Eosinophils infiltration was significant in group 5. After the treatment with Ful, TGF-β1 expression did not change and IL-6, IL-17A expressions were decreased in lung of the mice that inhaled OVA. It was not enough for Ful to relieve the neutrophil aggregation and improve inflammatory reaction in lung., Conclusion: The expressions of TGF-β1, IL-6, IL-17A in lung of the mice with OVA allergy were increased markedly after they inhaled specific antigen, which caused serious inflammation that was induced by neutrophil infiltration in lung. Ful could decrease the expressions of IL-6, IL-17A to some extent, but it was not enough to improve pathologic state in lung.

The objective of this work is to investigate how dilution rate and carbon-to-nitrogen (C/N) ratio affects lipid accumulation by Rhodosporidium toruloides AS 2.138 9 in continuous culture. Under steady-state conditions, the increase in dilution rate led to the decrease in lipid content and lipid yield. The highest lipid yield and lipid content at D = 0.02 h(-1) were 0.18 g lipid/g sugar and 57.1%, respectively, while the highest lipid productivity and biomass productivity were obtained at D = 0.14 h(-1). The increase in C/N ratio led to the increase in lipid content. The highest lipid content of 38% was obtained at C/N = 237. The highest lipid yield of 0.12 g lipid/g sugar was obtained at C/N = 92. However, the highest lipid productivity of 0.12 g/(L x h) was obtained at C/N = 32. No significant changes were observed in terms of fatty acid composition of the lipid produced under different C/N ratios, and these three fatty acids, palmitic acid, stearic acid and oleic acid, took over 85% in all samples.