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Objectives: To analyze the rules of acupoint selection in treatment of cancer-related insomnia with acupuncture and moxibustion by data mining technology., Methods: The articles of cancer-related insomnia treated with acupuncture and moxibustion were searched from CNKI, Wanfang, VIP, SinoMed, PubMed, WOS, Cochrane, and Embase databases, from the inception of each database to January 5, 2024. The prescription database of acupuncture and moxibustion for cancer-related insomnia was established. The descriptive analysis was conducted on the use frequency, meridian tropism and distribution of acupoints. Using SPSS Modeler 18.0 Apriori algorithm, the association rules of acupoint prescriptions were analyzed. With Cytoscape3.9.1 software used, the complex network diagram was plotted, and the cluster analysis of high-frequency acupoints was performed by SPSS26.0 software., Results: Forty-one articles were included, and 67 prescriptions were extracted with 89 acupoints involved, and the total use frequency was 447 times. The top 4 acupoints of the high use frequency were Baihui (GV20), Sanyinjiao (SP6), Shenmen (HT7) and Shenting (GV24). The included meridians were the governor vessel, the spleen meridian, the bladder meridian, the conception vessel, the heart meridian and the stomach meridian. The selected acupoints were mostly distributed on the head, the neck and and the upper and lower limbs. The special acupoints of the high use frequency included the five- Shu points, the crossing points and yuan -primordial points. Regarding acupoint combination, GV24, SP6, HT7, and GV20 were highly correlated. The three effective clusters were categorized among the top 12 acupoints of the high use frequency., Conclusions: In treatment of cancer-related insomnia with acupuncture and moxibustion, the principle focuses on supporting the healthy qi , eliminating pathogens, regulating yin and yang , promoting the circulation of the governor vessel for regulating the spirit, and tranquilizing the mind. The core acupoint prescription may includes GV24, SP6, HT7 and GV20；combined with Zusanli (ST36) and Yintang (GV4 + ) to enhance the therapeutic effect.

Objective: To describe the relapse status of smokers aged ≥15 years in China, and investigate the main factors influencing their relapse behavior. Methods: The data of this study cames from the 2018 China Adult Tobacco Survey, covering 400 committees or villages in 200 districts or counties in 31 provinces (autonomous regions and municipalities) of China. The relevant data of smoking and relapsing in residents aged ≥15 years were collected by face-to-face interview. Software SAS 9.4 was used to clean and analyze the data, and logistic regression model was used to analyze the influencing factors for relapse rate. Results: A total of 19 376 questionnaires were completed, with a response rate of 91.50%. In 2018, 66.05% of smokers aged ≥15 years in China had smoking relapse, in whom 66.59% were males and 55.79% were females. In all age groups, the age group 15-24 years had the highest smoking relapse rate (82.63%). Multivariate analysis showed that the younger age 15-24 years ( OR =4.618,95% CI :1.981-10.763), e-cigarette use ( OR =9.782,95% CI :3.139-30.490), and tobacco advertising, promotion and sponsorship in the past 30 days ( OR =1.710,95% CI :1.291-2.265) were associated with higher smoking relapse rate. Compared with people who were allowed smoking at home or those without smoking limit, the smoking relapse rate in people who were not allowed to smoke at home ( OR =0.562, 95% CI : 0.439-0.719) or those with smoking limit ( OR =0.487, 95% CI : 0.366-0.647) was lower. Conclusion: The smoking relapse rate in Chinese smokers is high, especially in young people. It is suggested to conduct targeted intervention based on the results of this study to reduce the smoking relapse rate and help achieve the smoking control goal in Healthy China 2030.

Objective: To observe the effect of electroacupuncture(EA) on the expression of nuclear transcription factors κB (NF-κB) and Tau protein and content of amyloid (Aβ) in diabetic rats with cognitive impairment, so as to explore its mechanism underlying improvement of learning-memory ability., Methods: Male SD rats were randomly divided into normal control ( n =9), model ( n =9) and EA ( n =9) groups. The diabetic mellitus model was established by feeding the rats with high fat and high sugar for 1 month and intraperitoneal injection of STZ (25 mg·kg-1·d-1) for 2 days. EA was applied to "Zusanli"(ST36) and "Neiting"(ST44) for 20 min, alternatively on both side every day, and "Yishu"(EX-B3) was stimulated by twirling the acupuncture needle with uniform reinforcing-reducing method for 1 min, followed by retaining it for 20 min. The treatment was conducted 6 times a week for 4 weeks. The learning-memory ability was evaluated by using Morris water maze swimming test. The blood glucose level was randomly detected by using a glucometer, the content of Aβ1-42 in the hippocampal tissue was detected by ELISA, and the relative expression levels of hippocampal Tau and NF-κB p65 proteins and mRNAs were determined by Western blot and fluorescence quantitative real-time PCR, separately., Results: After modeling, the blood glucose, escape latency, Aβ1-42 content and the expression levels of Tau and NF-κB p65 proteins and mRNAs in the model group were significantly increased ( P <0.01, P <0.05) in comparison with the normal control group. Following EA intervention, the modeling induced increase of blood glucose, escape latency, Aβ1-42 content and the expression levels of Tau and NF-κB p65 proteins and mRNAs were reversed ( P <0.05, P <0.01)., Conclusion: EA can improve the learning-memory ability in rats with diabetic cognitive impairment, which may be related to its function in down-regulating the levels of hippocampal Aβ1-42, Tau and NF-κB proteins.

Professor YANG Jun 's clinical experience of acupuncture and moxibustion for oculomotor paralysis is summarized. Professor YANG Jun pays attention to disease differentiation and syndrome differentiation in the treatment of this disease. According to the characteristics of oculomotor paralysis, "early diagnosis and seeking treatment from the source" is advocated. According to the etiology and pathogenesis, professor YANG divides oculomotor paralysis into three types: the syndrome of wind-evil attacking collaterals, the syndrome of spleen-stomach weakness and the syndrome of qi -deficiency and blood-stasis. As such, the acupoints are selected according to syndrome differentiation, and several different acupuncture methods (pricking needling at eyelids, penetrating needling and lifting eyelids and contralateral- balance needling on the healthy side) are adopted to improve the symptoms of oculomotor paralysis. It is also suggested to use the combination of scalp acupuncture and electroacupuncture to achieve the best dose-effect state. Moreover, local stimulation around the eyes is important to achieve the effects of " qi reaching affected area".

Objective: To investigate the exposure to tobacco advertisements and promotions among Chinese adults aged 15 and above, identify the publicity channels and forms of tobacco advertising and promotion in 2010 and 2018, and implicate further tobacco control measures in China. Methods: A multistage, stratified, randomized cluster sampling design was used in 2010 and 2018 China Adult Tobacco Survey, with national representativeness. 13 354 and 19 376 permanent residents were selected in the 2010 and 2018 surveys. SAS 9.4 software was applied for data analysis, and all the data were weighted based on a complex sampling design. Rao Scott χ 2 test was used for group comparison of a single factor. Results: In 2010 and 2018, 19.61% and 18.14% of the survey respondents did see tobacco advertising and promotion. From 2010 to 2018, there was no significant change in the situation of tobacco advertisements and promotions. Among those who had gone to a cigarette shop, the proportion of seeing tobacco advertisements increased from 29.28% in 2010 to 43.28% in 2018. Among those who had seen tobacco advertisements on TV, the rate fell from 50.93% in 2010 to 28.58% in 2018. Among those who had gone to movie theaters, the proportion of seeing tobacco advertisements increased from 2.17% in 2010 to 9.89% in 2018. Among those who had used the Internet, the proportion of seeing tobacco advertisements online rose from 19.20% in 2010 to 42.30% in 2018. In terms of tobacco promotion, the percentages of people who had seen tobacco promotions, cigarette price discounts, cigarette discount coupons, gifts, and other preferential activities in various places in the past 30 days were 4.99% vs. 9.30%, 0.78% vs. 4.09%, 0.04% vs. 0.33% and 0.98% vs. 3.33% in 2010 and 2018, respectively ( P <0.001). Conclusions: Tobacco advertising and promotion are still prevalent in China, with no significant change in 2010 and 2018. Tobacco advertising and promotion have been with the constant changes and development of media platforms. It is necessary to improve the implementation of relevant policies, comprehensively ban tobacco advertisements and promotions, and strengthen the supervision of ads and promotions.

Objective: To investigate secondhand smoke (SHS) exposure among non-current smokers aged 15 and over and its influencing factors in China in 2010 and 2018. Methods: The 2010 and 2018 China Adult Tobacco Surveys used multistage stratified cluster random sampling methods to obtain national representative samples across 31 provinces (autonomous regions and municipalities) in China. This study selected non-current smokers aged 15 and over as the research subjects to describe the general demographic characteristics, perceptions of SHS hazards, attitudes towards smoking bans in indoor areas in public places, and SHS exposure and the smoking restriction regulations in different places. The Rao-Scott χ 2 test was used to compare the rates, and the unconditional logistic regression was used to analyze the influencing factors of SHS exposure. All the subjects in the analysis were weighted based on a complex sampling design. Results: Among non-current smokers aged 15 and over, from 2010 to 2018, the percentage of indoor smokers had decreased from 84.7% to 71.9%. The rates of people who saw smoking in various places declined with a statistically significant difference ( P <0.001). The exposure to SHS for females, people aged 45-64, teachers, medical staff, workers in enterprises, businesses, waiters, and people who live in rural and central areas declined. The differences were statistically significant ( P <0.05). In 2018, the percentage of SHS exposure for those who reported a comprehensive smoking ban in households or allowed smoking in certain areas was lower than for those who reported allowed or without a smoking ban. Among those aged 25-44, SHS exposure for those who reported complete smoking prohibition in workplaces was lower than those who reported allowed or without smoking prohibition ( OR =0.65, 95% CI : 0.49-0.87). The proportion of SHS exposure for those who believe that smoking should not be allowed in indoor areas of 0-5 public places is higher than that smoking should not be allowed in the indoor spaces of 8 public places ( OR =2.13, 95% CI : 1.35-3.36). Among people aged 45-64, the proportions of SHS exposure for teachers ( OR =0.37, 95% CI : 0.17-0.78) and medical staff ( OR =0.35, 95% CI : 0.16-0.76) and staff working in agriculture, forestry, animal husbandry, fishery, and water conservancy were lower than governmental staff ( OR =0.49, 95% CI Data from the decreased exposure of Chinese non-current smokers aged 15 and over to SHS suggests the initial achievement through constructing a smoke-free environment, but continued efforts are needed. It is necessary to actively advocate for smoke-free families, strengthen publicity and education on the hazards of tobacco and SHS, and improve the skills of non-smokers in rejecting SHS exposure.Conclusions: Data from the decreased exposure of Chinese non-current smokers aged 15 and over to SHS suggests the initial achievement through constructing a smoke-free environment, but continued efforts are needed. It is necessary to actively advocate for smoke-free families, strengthen publicity and education on the hazards of tobacco and SHS, and improve the skills of non-smokers in rejecting SHS exposure.

Objective: To describe the current situation on quit intention and quit attempts among current smokers aged 15 years and above in China and explore the main factors affecting their smoking cessation behaviors. Methods: The subjects were people aged 15 years and above in China. A stratified multi-phased randomized cluster sampling method was used. The survey covered 200 districts/counties in 31 provinces (autonomous regions and municipalities directly under the central government), including 19 376 people under investigation. The data were weighted with SAS 9.4 for complex sampling analysis, and the frequency and weighted component ratio were used for description. The influencing factors were studied using the Rao Scott χ 2 test and unconditional multivariate logistic regression. Results: Among current smokers, 6.63% reported that they planned to quit smoking in the next month, 5.44% for men and 8.49% for women. 17.96% of the current smokers made quit attempts in the past 12 months, with 17.80% men and 22.15% women. People who smoke occasionally ( OR =3.42, 95% CI : 2.09-5.59) made quit attempts in the past ( OR =4.91, 95% CI : 0.33-0.52) and knew that smoking would lead to three diseases ( OR =2.24, 95% CI : 1.48-3.40) had higher quit intention. People who occasionally smoke ( OR =1.68, 95% CI : 1.20-2.34) received smoking cessation suggestions from medical staff ( OR =1.96, 95% CI : 1.62-2.37), were a ware of that smoking causes three diseases ( OR =1.63, 95% CI : 1.32-2.02) and had smoking prohibition regulations in indoor areas of their homes, had a higher rate of quit attempts. Conclusion: Providing effective smoking cessation intervention services and creating a supportive tobacco control environment are significant in promoting smoking cessation behavior.

Objective: To observe the effect of electroacupuncture(EA) on the expression of peroxisome proliferator-activated receptor gamma coactivators-1-alpha (PGC-1α), Irisin and brain-derived neurotrophic factor (BDNF) in the ischemic peripheral cortex, hippocampus and local skeletal muscle in rats with focal cerebral ischemic/reperfusion injury (CI/RI), so as to explore its underlying mechanism of improving of CI/RI., Methods: Male SD rats were randomly divided into 3 groups: sham-operation, model and EA (11 rats in each group). The focal CI/RI model was established by middle cerebral artery occlusion (MCAO). EA (2 Hz /15 Hz, 2 to 4 mA) was applied to "Quchi" (LI11) and "Zusanli" (ST36) of the affected side for 20 min, once a day for 7 days. Zea-Longa's score and Balance Beam score were used to evaluate the neurological and motor functions. The infarcted volume of the brain was detected by using 2,3,5-triphenyltetrazolium chloride staining. The expression levels of PGC-1α, fibronectin type III domain-containing protein 5(FNDC5) and BDNF proteins in the ischemic peripheral cortex, hippocampus and local skeletal muscle were detected by Western blot., Results: Compared with the sham-operation group, the Zea-Longa's score, Balance Beam score, percentage of cerebral infarct volume were notably increased ( P <0.01), while the expression levels of PGC-1α, FNDC5 and BDNF proteins in the cerebral cortex and hippocampus (not in the local muscle) were significantly down-regulated in the model group ( P <0.01, P <0.05). In comparison with the model group, the increase of Zea-Longa's score, Balance Beam score, percentage of cerebral infarct volume, and the decrease of expression levels of PGC-1α, FNDC5 and BDNF proteins in the ischemic peripheral cortex and that of BDNF in the hippocampus were reversed in the EA group ( P <0.01, P <0.05). No significant changes were found in the expression levels of hippocampal PGC-1α and FNDC5 proteins in the hippocampus and those of PGC-1α, FNDC5 and BDNF proteins in the local muscle after EA intervention ( P >0.05)., Conclusion: EA can improve neurological and motor functions and reduce cerebral infarction volume in CI/RI rats, which may be related to its functions in activating PGC-1α/Irisin(FNDC5)/BDNF pathway in the cerebral cortex.

A 43-year-old male patient with onset of edema caused by nephrotic proteinuria and low titer of anti-M type phospholipase-A 2 -receptor (PLA 2 R) antibody was diagnosed as idiopathic membranous nephropathy by renal biopsy. Administrated with prednisone 40 mg once a day and cyclosporine 100 mg twice a day as front-line regimen, the patient relapsed after transient partial remission. When treatment was combined with cyclophosphamide 100 mg once a day, the 24-hour total urine protein and titer of anti-PLA 2 R antibody were even elevated. Therefore, the patient received rituximab 1 g intravenously in April 2019, October 2019 and October 2020 respectively. CD19 positive B lymphocytes in peripheral blood were eliminated from 71/μl to zero. Immunosuppressants and corticosteroids were withdrawn successively. On the last follow-up in November 2020, the anti-PLA 2 R antibody was negative, and the 24-hour total urine protein and serum albumin was 4.4 g and 34 g/L, respectively. This case suggested the potential efficacy of rituximab for refractory membranous nephropathy. Further studies should explore whether the titer of anti-PLA 2 R antibody indicates the dose of rituximab.

To summarize professor YANG Jun 's experience in treatment of refractory facial paralysis by different acupuncture manipulation. Professor YANG Jun believes that refractory facial paralysis has prolonged course, qi and blood deficiency and collaterals injury after a long illness, and it is difficult to obtain obvious curative effect with routine acupuncture. Aimed at the etiology, pathogenesis and characteristics of disease, professor YANG Jun adopts 3 kinds of acupuncture manipulations, such as "penetration needling with long needle" "pulling technique exert after needle stuck" "contralateral-balance needling on the healthy side", to relieve deviation.

Onset with fever and back pain, an 81-year-old man had sudden oliguria and progressively elevated serum creatine from normal range to 660 μmol/L within 1 week after receiving contrast agents, various antibiotics, and several nonsteroidal anti-inflammation drugs. Urine output recovered after supportive treatment. However, his serum creatinine level rose again soon after a temporary decline accompanied by gross hematuria with almost normal morphology, nephrotic proteinuria, and hypoalbuminemia. Renal biopsy revealed necrotizing glomerulonephritis. Methylprednisolone was intravenously administrated 500 mg per day for 3 days, followed by oral glucocorticoids and cyclophosphamide. Gradually the patient's serum creatinine descended to 144 μmol/L.

Objective: To compare the results of visual acuity testing for teenagers with visual acuity charts designed at 2.5-meter and 5-meter distances, and to investigate the accuracy of the 2.5-meter visual acuity chart. Methods: It was a self-control study. A total of 227 teenagers (454 eyes) with ametropia who came to the ophthalmic clinic of Peking Union Medical College Hospital from March 2019 to September 2019 were included. There were 123 males and 104 females aged (11.74±3.26) years. The vision examiners were trained in the same testing environment and passed the consistency test. Distance visual acuity of all participants was tested with charts designed at 2.5 meters and 5 meters in a 10-minute interval. According to the age (7-9, 10-12, 13-15 and 16-18 years old) and visual acuity (1.00-0.52, 0.40-0.30 and 0.22-0.10), the results of two kinds of visual acuity charts were compared. The testing order of the two visual charts was randomly determined. The visual acuity results were converted into logMAR values and compared. Paried t -student test was used to compare the difference between two charts, and Pearson correlation test was used to explore the correlation between the results of two charts. Results: The visual acuity of the right eye was 0.37±0.24 (logMAR) at 2.5 meters and 0.50±0.26 (logMAR) at 5 meters. The distance visual acuity measured with the 2.5-meter visual acuity chart was better ( t =16.19, P <0.01). The visual acuity of the left eye was 0.36±0.23 (logMAR) at 2.5 meters and 0.45±0.23 (logMAR) at 5 meters ( t =11.71, P <0.01). The differences between two charts were larger among teenagers with lower visual acuity ( r =0.387, P <0.01). Conclusion: Under the same test conditions, the distance visual acuity measured with a 2.5-meter chart was significantly better than a 5-meter chart. The visual acuity chart designed at 2.5 meters was not an appropriate tool to measure distance vision in adolescents. ( Chin J Ophthalmol, 2021, 57: 122-125) .

Objective: To observe the effect of moxibustion on cognitive function in patients with cognitive decline of type 2 diabetes mellitus (T2DM)., Methods: A total of 47 patients with cognitive decline of T2DM were randomized into an observation group (24 cases, 4 cases dropped off ) and a control group (23 cases, 3 cases dropped off ). Conventional hypoglycemic intervention was adopted in the control group. On the basis of the treatment in the control group, Huayu Tongluo moxibustion (moxibustion for removing stasis and clearing collaterals) was applied at Baihui (GV 20), Dazhui (GV 14), Shenting (GV 24), once every 2 days, 4 weeks were as one course, totally 3 courses were required. Before and after treatment, scores of auditory verbal learning test (AVLT), complex figure test (CFT), Montreal cognitive assessment (MoCA) and trail making test-A (TMT-A) were observed in the both groups, and the clinical effective rates were compared., Results: After treatment, the scores of immediate memory and delayed memory of AVLT in the observation group were higher than those before treatment and those in the control group ( P <0.05). After treatment, the scores of visual space and executive function, delayed memory, attention and orientation of MoCA in the observation group were higher than those before treatment and those in the control group ( P <0.05). The effective rate was 85.0% (17/20) in the observation group, which was superior to 10.0% (2/20) in the control group ( P <0.05)., Conclusion: Huayu Tongluo moxibustion has a definite effect on cognitive decline of type 2 diabetes mellitus.

To scientifically evaluate the intervention effect of Chinese medicine preventive administration(combined use of Huo-xiang Zhengqi Oral Liquid and Jinhao Jiere Granules) on community population in the case of coronavirus disease 2019(COVID-19), a large cohort, prospective, randomized, and parallel-controlled clinical study was conducted. Total 22 065 subjects were included and randomly divided into 2 groups. The non-intervention group was given health guidance only, while the traditional Chinese medicine(TCM) intervention group was given two coordinated TCM in addition to health guidance. The medical instructions were as follows. Huoxiang Zhengqi Oral Liquid: oral before meals, 10 mL/time, 2 times/day, a course of 5 days. Jinhao Jiere Granules: dissolve in boiling water and take after meals, 8 g/time, 2 times/day, a course of 5 days, followed up for 14 days, respectively. The study found that with the intake of medication, the incidence rate of TCM intervention group was basically maintained at a low and continuous stable level(0.01%-0.02%), while the non-intervention group showed an overall trend of continuous growth(0.02%-0.18%) from 3 to 14 days. No suspected or confirmed COVID-19 case occurred in either group. There were 2 cases of colds in the TCM intervention group and 26 cases in the non-intervention group. The incidence of colds in the TCM intervention group was significantly lower(P&lt;0.05) than that in the non-intervention group. In the population of 16-60 years old, the incidence rate of non-intervention and intervention groups were 0.01% and 0.25%, respectively. The difference of colds incidence between the two groups was statistically significant(P&lt;0.05). In the population older than 60 years old, they were 0.04% and 0.21%, respectively. The incidence of colds in the non-intervention group was higher than that in the intervention group, but not reaching statistical difference. The protection rate of TCM for the whole population was 91.8%, especially for the population of age 16-60(95.0%). It was suggested that TCM intervention(combined use of Huoxiang Zhengqi Oral Liquid and Jinhao Jiere Granules) could effectively protect community residents against respiratory diseases, such as colds, which was worthy of promotion in the community. In addition, in terms of safety, the incidence of adverse events and adverse reactions in the TCM intervention group was relatively low, which was basically consistent with the drug instructions.

A refractive index insensitive temperature sensor is proposed base on cascading single mode fiber with few mode fiber(FMF). During the sensor preparation, the splicing current is set to 100 mA, and a section of FMF is no core-offset splicing between two single-mode fibers. Therefore, it can motivate the transmission mode preferably and form optical fiber Mach-Zehnder interferometer. The mode phase difference in FMF will be changed according to the outside environment. It will cause interference fringe shift. The parameter to be measured can be achieved by detecting the amount shift of interference spectrum. The FMF can transmit four modes with LP01, LP11, LP21, LP02. The transmission spectrum is also analyzed, which shows that they have two modes of LP01 and LP11 in sensor with the length of 81.5 mm. In the refractive index and temperature sensing experiment, the cascading FMF sensor with the length of 81.5 mm is used. The results show that the transmission spectrum of sensor appears obvious blue shift as temperature is increasing, the temperature sensitivity can be up to -85.9 pm·℃-1 within the range of 27.6~93.8 ℃ with good linearity. The refractive index sensitivity is 3.697 34 nm·RIU-1 within the range of 1.347 1~1.443 9. There is no obvious shift phenomenon in the transmission spectrum with the feature of refractive index insensitive. Therefore, compared with the traditional cladding mode and multimode interferometric fiber-optic sensor, the proposed sensor based on FMF is easier to control and analyze transmission mode has the advantages of simple structure, easy process and high sensitivity. It can avoid cross-sensitivity between temperature and refractive index measurement. Thus, it can be used for temperature detection of power system, biomedicine, aerospace and other fields.

This study aimed to analyze the effect of temperature on performance and microbial community structure of an anaerobic packed bed reactor (APBR). The temperature was increased step-wise from room temperature (22 degrees C ± 1 degrees C) to psychrophilic (15 degrees C ± 1 degrees C), mesophilic (37 degrees C ± 1 degrees C) and thermophilic (55 degrees C ± 1 degrees C). The results showed that, in the temperature changing process, the higher the temperature of APBR was, the higher COD removal rate and daily gas production were. After temperature changed to psychrophilic, mesophilic and thermophilic, COD removal rate and daily gas production were 25%, 45%, 60% and 2.3 L x d(-1), 4.0 L x d(-1), 8.5 L x d(-1) respectively. However, there was no significant change in biogas composition (-60%). A sudden temperature change caused a simultaneous increase in the concentration of volatile fatty acids (VFA), which had been fluctuating. Using 16S rRNA gene clone library screening, Euryarchaeota was commonly found, including important methanogens: MBT (Methanobacteriales), Mst (Methanosaetaceae) , Msc (Methanosarcinaceae) and MMB (Methanomicrobiales), as well as thermophilic bacteria and few spring Archaea. However, the diversity of methanogenic groups was reduced, especially at mesophilic. The results of quantitative PCR showed that the 16S rRNA gene concentrations of Mst, MMB and Msc were reduced by temperature changes. Although the relative proportion of every kind of methanogen was significantly affected, Mst was the dominant methanogen.

Objective: To investigate the prognostic factors for non-small cell lung cancer(NSCLC)in patients under 40 years of age., Methods: The clinicopathological data of 148 young patients with NSCLC were retrospectively analyzed. Kaplan-Meier and Cox regression analyses were used to analyze the relationship between prognostic factors and survival time., Results: The patients were followed-up for 6 - 148 months, and the follow-up rate was 100%. In the whole group, 122 patients died and 26 cases were surviving. The 1-, 3- and 5-year survival rates were 54.7%, 10.4% and 5.6%, respectively. The median survival time (MST) was 14.7 months. Kaplan-Meier analysis showed that Karnofsky performance status (KPS), clinical stage, treatment modality and serum CEA were related with prognosis (P < 0.05). Multivariate analysis indicated that KPS, clinical stage, treatment modality and serum CEA were independent prognostic factors (P < 0.05)., Conclusions: KPS, CEA, clinical stage and treatment modalities are independent prognostic factors in young NSCLC patients.

Objective: To observe the immune response of Th17 cells in mesenteric lymph node (MLN) of C57BL/6 mice infected by Schistosoma japonicum., Methods: Twenty C57BL/6 mice were randomly divided into infected group and control group each with ten mice. The mice in infected group were infected each with 40 +/- 5 S. japonicum cercariae. Five to six weeks later, MLN lymphocytes were separated and stimulated for 4 h by anti-CD3 (1 microg/ml) and anti-CD28 (1 microg/ml) before examination of IL-17 and retinoic acid receptor-related orphan receptor gammat (ROR-gammat) mRNA by reverse transcription PCR. The level of IL-17 and IFN-gamma was detected by ELISA after culturing with supernatant for 72h. MLN lymphocytes were stimulated for 5h by 10 ng/ml phorbol myristoyl acetate (PMA) and 1 microg/ml ionomycin. The intracellular cytokines were stained and the content of Th17 and other cytokines was examined by flow cytometry., Results: The level of IFN-gamma [(214.3 +/- 62.6) pg/ml] and IL-17 [(176.8 +/- 62.1) pg/ml] in the supernatant of cultured MLN cells from the infected mice was significantly higher than that of normal mice [(467 +/- 13.9) and 0 pg/ml) (P < 0.05). The expression level of IL-17 and ROR-gammat mRNA was also considerably higher than that of normal mice. IL-17+ IL-4+, IL-17+ IFN-gamma+, IL-17+ IL-5+ and IL-17+ IL-9 cells accounted for 0.06%, 0.02%, 0.02%, and 0.01% of the mesenteric lymph node CD4+ T cells of the infected mice, respectively. However, IL-17+ IL-10+ and IL-17+ Foxp3+ cells were undetected., Conclusion: The MLN of S. japonicum-infected C57BL/6 mice can induce the production of Th17 cells, and these cells can secrete IL-4, less IFN-gamma, IL-5 and IL-9, but not IL-10, and can not express Foxp3 in the infected mice.

Objective: To investigate the risk perception of heat wave, and further explore its related factors in Guangdong province., Methods: A total of 2183 adults were selected by a multi-stage sampling method in Guangdong province. Each subject was interviewed in their home with a structured questionnaire by a well trained investigator from September to November, 2010. The questionnaire contained socio-demographic characteristics, heat wave related knowledge, risk perception of heat wave, etc. Chi-square test and multivariate logistic regression were employed in this study., Results: The average age of total 2183 participants was (39.31 ± 14.16) years, among which 53.37% (1165/2183) were males, and 48.74% (1064/2183) were selected from urban. About 38.11% (832/2183) of participants heard about heat wave, and 38.52% (841/2183) of subjects thought the heat wave had higher impact on their health (risk perception score of heat wave ≥ 5 points). About 81.91% (1788/2183) of all participants thought the weather in most recent years was hotter than several years ago. Among these people, 30.48% (545/1788) thought the main reason of weather becoming hotter was due to emission of carbon dioxide, and 26.51% (474/1788) thought it was due to air pollution. Results from the multivariate logistic regression showed that the risk perception score of heat wave were higher in subjects with higher education (OR = 2.16, 95%CI: 1.41 - 3.30), from urban(OR = 1.37, 95%CI: 1.10 - 1.72), with higher score of trust(OR = 1.08, 95%CI: 1.01 - 1.14) and participants with higher score of heat wave related knowledge (OR = 1.39, 95%CI: 1.27 - 1.52). Furthermore, compared to hierarchist, egalitarian (OR = 1.73, 95%CI: 1.30 - 2.29), individualist (OR = 1.93, 95%CI: 1.41 - 2.65) and fatalist (OR = 1.80, 95%CI: 1.41 - 2.29) also had higher risk perception score of heat wave., Conclusion: There is a lack of knowledge and risk perception to heat wave among the residents in Guangdong province. Risk perception of heat wave was higher in people who were from urban, had higher level of trust on government, experts and media, had higher health-related knowledge score, and non-hierarchists.

Objective: To compare docetaxel plus prednisone with mitoxantrone plus prednisone as first-line chemotherapy for metastatic hormone-refractory prostate cancer (mHRPC)., Methods: From January 2007 through August 2010, 62 patients with mHRPC received 5 mg of prednisone twice daily were randomly assigned to receive mitoxantrone 12 mg/m² every three weeks (group A) or 75 mg/m² every three weeks (group B). The cycles of each regimen were less than 10 times. The primary end point was overall survival. The secondary end points were the prostate-specific antigen (PSA) response rate, the duration of PSA response and the objective tumor response rate (ORR). All the t test, χ² test and Fisher's exact test were performed between 2 groups., Results: Thirty-one patients enrolled in group A received a median 4 cycles of regimen (range 1 - 10), whereas 30 patients enrolled in group B received a median of 7 cycles of regimen (range 2 - 10). There were 45.2% patients in group A and 70.0% in group B had PSA response (χ² = 3.85, P < 0.05). The duration time of PSA response was 121 days (range 20-323 days) in group A and 168 days (range 42 - 447 days) in group B, respectively. The ORR was 15.0(3/20) in group A and 10.3% (3/29) in group B, respectively. The median survival was 511 days (95%CI: 357 - 665 days) in group A and 833 days (95%CI: 634 - 1032 days) in group B, respectively (χ² = 4.20, P = 0.040). The incidence of thrombocytopenia in group A was higher than group B (χ² = 5.60, P = 0.018); the incidences of nausea and vomiting (χ² = 4.32, P = 0.038), diarrhea (P = 0.024), fatigue (χ² = 5.90, P = 0.015), and alopecia (χ² = 5.42, P = 0.020) in group B were higher than group A., Conclusion: Docetaxel plus prednisone can lead to superior overall survival and PSA response rate in patients with mHRPC.

Objective: To explore the relationship between distorted weight perception and suicide ideation among normal weight adolescents in Guangdong province., Methods: This study used the data of Guangdong Provincial Youth Health Risk Behavior Survey in 2004 and 2007. To identify the association between distorted weight perception and suicide ideation, a logistic regression analysis was performed. The effects of age, economic status, mother's education, mental health, and depression were also adjusted., Results: A total of 12 729 people participated in this study, 6096 males and 6633 females. The prevalence of suicide ideation among normal weight adolescents was 13.58% (1729/12 729) in the past 12 months, with higher prevalence in girls (16.15%, 1071/6633) than that in boys (10.79%, 658/6096) (χ(2) = 77.71, P = 0.00). It was common that the adolescents misperceived their weight. Only 44.93% (5719/12 729) of normal weight students correctly perceived their body weight while 43.52% (5540/12 729) of them overestimated their weight and 11.43% (1455/12 729) underestimated their weight. The distorted weight perception in girls (65.58%, 4350/6633) was higher than that in boys (43.39%, 2645/6096) (χ(2) = 993.91, P = 0.00). Distorted weight perception was significantly associated with suicide ideation after controlling for factors age, economic status, mother's education, mental health, and depression. The students who overestimate their body weight were more likely to have suicide ideation than that who correctly perceived their weight (OR = 1.43, 95%CI: 1.27 - 1.61)., Conclusion: The prevalence of distorted weight perception was high and it significantly associated with suicide ideation.

Objective: To compare the magnitude of inequities in health-related behaviors among males in Guangdong province, and to investigate the extent of the disparities., Methods: Data sets available from the Guangdong Chronic Diseases and Risk Factors Survey 2007 are used. Concentration index (C) and concentration curve are employed to measure the differential of males' health-related behaviors across urban and rural areas in Guangdong. Odds ratios of 6 health-related behaviors among different areas are derived from 4 logistic models, after adjusting for age, married state, educational status, occupation and income., Results: Results from Cs reveal that the inequality gradients disadvantageous to men in rural areas are:smoking (C = -0.075, P = 0.000), alcohol intake (C = -0.023, P = 0.002), blood pressure (C = 0.106, P = 0.000), blood sugar (C = 0.114, P = 0.000) and weight (C = 0.107, P = 0.000), while lack of physical activity (C = 0.044, P = 0.000)concentrates in the more affluent areas. The magnitudes of these inequalities appear to be higher on health-seeking behaviors than on health-risk behaviors. After adjusting for age and marital status, there is still strong evidence showing the rural-urban differences in the health related behaviors among males in Guangdong province. When educational status, occupation and income are added to the logistic model as control factors, the results have led to a loss of statistical significance on such rural-urban inequalities, indicating that socioeconomic factors play an important role on these health-related behaviors which leads to the inequalities among males in Guangdong province., Conclusion: To reduce the gaps in health-related behaviors seen in the rural and urban areas, effective policies should be developed to change the social determinants of rural-urban differences in health and to strengthen the implementation of health-related programs on those vulnerable groups.

Objective: To investigate the factors involved in the development of retinopathy of prematurity (ROP), and to provide the preliminary data for the evaluation of current criteria for ROP screening., Method: Premature infants with birth body weight (BBW) ≤ 2000 g or gestational age (GA) ≤ 34 weeks in the two hospitals in Zhejiang between March 2005 and November 2008 were recruited and examined by indirect ophthalmoscopy. The records were analyzed., Result: One thousand two hundred and twenty-five premature infants were included. Of them, 713 were male and 512 female. There were 179 twins and 21 triplets in the premature infants. The incidence of ROP was 10.8% (132 in 1225 patients). There were 12 cases (0.98%) to the point of pre threshold ROP. 4 cases (0.3%) developed threshold ROP. Only one case developed pre threshold ROP of low risk among 65 cases without history of oxygen treatment (1.5%). The percentage has significant difference compared to that of cases with history of oxygen (χ(2) = 5.115, P < 0.01).Between ROP and Non-ROP groups, there was significant difference in BBW(F = 26.39, P < 0.001), gestational age (F = 19.73, P < 0.001), but there was no significant difference in sex (χ(2) = 0.279, P > 0.05) or twins and triplets (χ(2) = 3.449, P > 0.05). The incidence of ROP among premature infants with BBW ≤ 1000 g was more than three times of that with BBW > 1000 g, and the incidence of ROP among premature infants with GA ≤ 28 weeks was about 2.5 times of that with GA > 28 weeks. Logistic regression analysis indicated that less BBW or shorter GA or undulation of blood oxygen concentration was a significant risk factor involved in the development of ROP (r = 0.57, P < 0.05). All ROP patients were cured., Conclusion: Less BBW, shorter GA and undulation of blood oxygen concentration are the important risk factors for the development of ROP. Premature infants with BBW ≤ 1000 g or GA ≤ 28 weeks, who had oxygen history, should be given very special attention in the ROP screening. The current criteria for ROP screening should be narrowed. In general, the ROP screening has lowered the incidence of blindness among children by investigating and treating ROP timely.

Objective: To study the protective immunity induced by recombinant vaccination of Cs-Rho GTPase of Clonorchis sinensis (Cs)., Methods: 20 SD-rats(8 weeks) were divided into two groups: A (recombinant protein experiment group) and B (PBS control group). Rats in group A were immunized with 1 ml protein of Cs-Rho GTPase (90 microg/ml) and 1 ml Freund's complete adjuvant through back and vola. 2 week later, the rats were given 1 ml protein of Cs-Rho GTPase (90 microg/ml) and 1 ml Freund's incomplete adjuvant, followed by 1 ml protein of Cs-Rho GTPase (90 microg/ml) through intraperitoneal injection at 4, 7, 11 week after the first immunization. Rats in group B were given PBS in the same way as group A. All rats were challenged each with 50 Clonorchis sinensis metacercariae after the last immunization. 21 d later, fecal samples were collected from all rats for examining eggs (number of eggs per gram feces, EPG) in every 3-5 d. When eggs were found, the rats were sacrificed and worms were collected. IgG, IgG1 and IgG2a in sera were detected by ELISA before every immunization. Mean number of worms and eggs, and antibody level in the experiment group were calculated and statistically compared with the controls., Results: The mean number of worms and EPG were (9.2 +/- 9.9) and (956.8 +/- 1 062.5) respectively in group A, which were significantly lower than those of group B [(23.25 +/- 15.75) and (3 062.5 +/- 2 501.8) respectively] (P < 0.05). The absorbency values of serum IgG (0.1, 0.45, 0.65, 0.6, 0.65), IgG1 (0.1, 0.45, 1.1, 1.0, 1.1), and IgG2a (0.1, 0.7, 1.1, 1.1, 1.1) before every immunization in group A were significantly higher than those of group B (almost always 0.1) (P < 0.05)., Conclusion: Recombinant vaccination of Cs-Rho GTPase induces partial protective immunity against Clonorchis sinensis infection in rats.

In the present paper, a laboratory-made high-performance electrophoresis microcolumn unit was prepared for UV-Vis spectrophotometer. X-ray diffraction was used in the preparation of electrophoretic microcolumns. And an analytical technique of microcolumn electrophoresis coupled with UV-Vis spectrophotometry was introduced. Uniform quartz microncrystals were prepared by hydrothermal synthesis. Their crystalline phase and morphology were identified by X-ray diffraction and scanning electron microscope, respectively. The quartz microncrystals were packed into a 2-mm i. d. fused-silica tube to prepare the electrophoretic microcolumn. With 1.5 mmol x L(-1) disodium phosphate buffer solution (pH 11.5) containing 25% (phi) methanol and 10% (phi) acetonitrile, tryptophan, phenylalanine and tyrosine were on-line separated on line and detected by microcolumn electrophoresis coupled with UV-Vis spectrophotometry without derivatization. The limits of detection were 0.037, 0.20 and 0.20 micromol x L(-1), respectively. The separation efficiency of tryptophan was 4.5 x 10(4) plates/m. The sample capacity of the electrophoretic microcolumn achieved 35 microL. It was found that the electrophoretic microcolumn packed with quartz microncrystals was able to limit Joule heat, increase sample capacity and enhance detection sensitivity. The laboratory-made electrophoretic microcolumn could be a high-performance separation unit for conventional UV-Vis spectrophotometer. The on-line coupling of microcolumn electrophoresis and UV-Vis spectrophotometry could separate and determine samples with complicated matrices, reduce zone broadening and enhance separation efficiency, so expand the analytical function of spectrophotometer in the trace analysis of mixed components with overlapped spectra.

Objective: To investigate whether there is an association between the expression of B7-H1 in HBV transgenic mice and the immune tolerance to HBV., Methods: T cells stimulatory capacity of DC was analyzed using mixed lymphocyte reaction. Expression of MHC-II, CD80, CD86, B7-H1 on DC was detected by Flow Cytometry. IL-2, IFNgamma, IL-10 production of T cells were determined by using ELISA. B7-H1 mRNA and protein expression in liver tissue were detected by RT-PCR and Western blotting respectively., Results: The ability of DC cells from HBV transgenic mice to stimulate T cell proliferation was significantly impaired compared with DC cells from control mice (t = 16.674, 19.674, 21.712, P less than 0.01). Expression of MHC-II, CD80 on DC was markedly decreased in transgenic mice (t = 7.910, 6.413, P less than 0.05). Meanwhile, the expression of CD86 and B7-H1 on DC cells in HBV transgenic mice were not significantly different from that in control mice. The levels of IL-2, IFNgamma, IL-10 in supernatant of T cells was significantly lower compared with controls (t = 18.712, 18.712 and 11.683, P less than 0.05). There was no significant difference in B7-H1 expression at mRNA and protein levels in liver tissue compared with controls., Conclusions: Functional defect of DC, partly due to decreased expression of MHC-II, CD80, but not related to B7-H1 expression, is the cause for immune tolerance to HBV in HBV transgenic mice.

Objective: To explore the feasible age limits in Chinese elderly patients with non-Hodgkin's lymphoma (NHL)., Methods: The clinical data of 507 patients with NHL who were admitted to Peking Union Medical College Hospital (PUMCH) from January 1990 to December 2007 were retrospectively analyzed. They were further followed up by reviewing medical records or by phone. The deadline of follow-up was October 2008., Results: The 5-year/8-year overall survival (OS) rates were 64.6%/45.7%, 53.0%/ 44.1%, 32.8%/17.5%, 40.0%/22.8%, and 19.8%/0, respectively, in patients aged < 60 years, 60-64 years, 65-69 years, 70-74 years, and > or = 75 years. The OS rate was significantly different between patients aged > or = 75 years and other age groups, and between patients aged 65-70 years and patients younger than 60 years (P < 0.05). Only age, serum albumin, and hemoglobin affected the survival status in elderly NHL patients., Conclusion: Sixty-five years can be regarded as the age limit in Chinese NHL patients.

Objective: To study the association of integrin alpha-2 (ITGA2) gene C807T, integrin beta-3 (ITGB3) gene T176C polymorphisms with ischemic stroke and the effect of the polymorphisms on plasma lipid and lipoprotein levels., Methods: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to detect the integrin genotypes in 265 patients with ischemic stroke and 280 healthy controls. The plasma lipid and lipoprotein levels were measured by routine method., Results: Plasma total cholesterol (TC), triacylglycerol (TG) and low density lipoprotein-cholesterol (LDL-C) in the patients with ischemic stroke were significantly higher than those in the controls (P< 0.05). The distributions of the ITGB3 gene T176C polymorphism were not different between the ischemic stroke group and control group, but the ITGA2 gene C807T polymorphism was significantly different. The relative risk suffering from ischemic stroke of the T allele carrier was 1.455 times as that of the C allele carrier (OR=1.455, 95%CI: 1.134-1.866). The level of plasma lipid in the T allele carriers was significantly higher than that in the C allele carriers (P< 0.05)., Conclusion: The ITGA2 gene C807T polymorphism was associated with ischemic stroke, the 807 T allele may be a genetic risk factor for ischemic stroke. The ITGA2 gene C807T polymorphism may affect ischemic stroke through plasma lipid and lipoprotein levels.

Objective: To investigate the effect of Cedemex on cAMP and cGMP contents in different brain regions in morphine withdrawal rats precipitated by naloxone., Method: A physical morphine dependent model of rats was established by subcutaneous injection of morphine in gradually increasing dosage within 7 days. cAMP and cGMP contents of VTA, cortex and hippocampus of the rat brains were determined by radioimmunoassay., Result: The morphine withdrawal symptoms of rats were relieved significantly by ig Cedemex. Compared with the controls, cAMP content in the region of VTA, cortex and hippocampus of the morphine dependent rats were significantly higher (P < 0.05), while cGMP contents in those regions were significantly lower (P < 0.05). cAMP contents in the area of VTA, cortex and hippocampus of the morphine dependent rats were significantly reduced, while cGMP contents were significantly increased by ig Cedemex., Conclusion: Cedemex may significantly attenuate the morphine withdrawal symptoms in rats. The mechanism of this effect may be related to adjusting the contents of cAMP and cGMP in some brain regions.

Objective: To detect the effect of gossypol, praziquantel and artemether on activity of the recombinant lactate dehydrogenase of Schistosoma japonicum (rSjLDH)., Methods: Effect of gossypol (0-0.10 mmol/L), praziquantel (0-0.20mmol/L) and artemether (0-0.10 mmol/L) on the enzymatic activity of rSjLDH was detected by spectrophotometric method in standard reaction system established before, same solvent of each reagent was used as control. Results were analyzed by SPSS13.0 software., Results: Compared with the control, considerable inhibition for both reduction and oxidation reactions catalyzed by rSjLDH was observed at 0.04 mmol/L of gossypol (P < 0.01). The enzyme activities of rSjLDH for reduction and oxidation reactions were inhibited by 91.3% and 89.1% respectively at 0.1 mmol/L of gossypol, com-pared with the control (P < 0.01). To praziquantel, enzymatic activity inhibition was observed at 0.02 mmol/L for oxidation reaction and at 0.06 mmol/L for reduction reaction (P < 0.05). At 0.2 mmol/L of praziquantel, enzymatic activities were inhibited by 83.8% for reduction reaction and 72.2% for oxidation reaction respectively, than that of the control (P < 0.01). No inhibition for both reduction and oxidation reactions was observed at 0.1 mmol/L of artemether, compared with that of the control (P > 0.05)., Conclusion: Gossypol and praziquantel show a high inhibition on rSjLDH. SiLDH may be one of the molecular targets of praziquantel.

Objectives: To study the expression of Polo-like kinase 1 (PLK1) and E-cadherin in the tissues of hepatocellular carcinoma, and to discuss the relationship between them and clinical-pathological features, and to evaluate their prognostic value of hepatocellular carcinoma after liver transplantation., Methods: mRNA and protein expression of PLK1, E-cadherin were detected by RT-PCR and immunohistochemistry method respectively, the correlations of them with clinical-pathological data, tumor free time, recurrence rate were compared and analyzed., Results: The mRNA expression was observed in 90.0% for PLK1 and 96.0% for E-cadherin, and higher in cancerous' tissues than paracancerous' of all cases for PLK1 but no trend for E-cadherin. The positive and decreased expression rate for PLK1 and E-cadherin was observed in 60.0% and 50.0% respectively, the positive PLK1 expression was correlated with preoperative serum alpha-fetoprotein (AFP) only (chi2 = 4.433, P = 0.035), while E-cadherin expression was associated with none of the clinical-pathological features. There was a correlation between the positive PLK1 and decreased E-cadherin expression (chi2 = 5.333, P = 0.021). PLK1 (P = 0.006), E-cadherin (P = 0.019) and larger tumor (P = 0.019), portal vein tumor thrombi (P = 0.030), Edmondson grading (P = 0.019), preoperative serum AFP (P = 0.020) were all correlated with recurrence rate under Kaplan-Meier analysis, while only PLK1 (RR = 3.104, P = 0.009) had significant difference under Cox regression analysis., Conclusions: The positive PLK1 expression and the decreased E-cadherin expression indicate higher recurrence rate of HCC after liver transplantation, and PLK1 is a independent risk factor.

In order to study the distributions characteristics, sources and relationship of PAHs in PM10- phoenix tree leaves-soil system of a coking & chemical factory in Shanghai, the samples of PM10, phoenix tree leaves and soil around the factory were collected for a year. The concentration of PAHs were analyzed according to the USEPA method 8 000 series. The results showed that the average concentration of PAHs in PM10, phoenix tree leaves and soil were 101.11 ng/m3, 79.45 ng/g and 121.53 microg/g, respectively. Particulate phase (PM10) contained mainly carcinogenic and mutagenic PAHs, among which BaA, BghiP, Flu and BaP were found at significant concentrations. In phoenix tree leaves, Nap,Chy, BaP and BghiP presented a higher level of concentration. In soil, 3 and 4-ring PAHs presented a higher level. PAHs concentrations of phoenix tree leaves were very lower in May. Only Ace (0.16 ng/g) and Pyr (0.63 ng/g) were detected. In July and August the concentrations (39.19 ng/g and 150.94 ng/g, respectively) were uplifted significantly. It could be concluded PAHs was from petroleum and coal-fired compound source. There were very strong positive relationships of 16 PAHs level among phoenix tree leaves, soil and PM10 (p < 0.01).

Objective: To screen cDNA library from Clonorchis sinensis for gene identification, and to clone and construct library of secreted recombinant proteins., Methods: cDNA library from Clonorchis sinensis was screened for identifying new genes by Blastn protocol, the sequence of which was further analyzed by Motifscan and NCBI Conserved Domain Search protocol., Results: The secreted proteins (Pcs004f03) were identified, with a 689 bp DNA sequence (187aa), and a theoretical molecular weight of Mr 21100. It is predicted that the gene CsCrSP contains one N-glycosylation site, three N-myristoylation sites, three casein kinase II phosphorylation sites, one cAMP-and cGMP-dependent protein kinase phosphorylation site and protein kinase C phosphorylation site, a signal peptide at N-terminals and SCP-like extracellular protein domain., Conclusion: The CsCrSP gene is a secretory cysteine-rich protein with a SCP-like extracellular protein domain.

Objective: To investigate the effects of ciclosporin (CsA) and tacrolimus (FK506) on replication of hepatitis B virus (HBV) in vitro., Methods: HBV genome permanently transfected human liver cancer cells of the line HepG2.2.15 were cultured. CsA and FK506 at different concentrations were added into the culture fluid so as to identify the nontoxic concentrations by MTT method. Then the HepG2.2.15 cells were treated by CsA and FK506 at different nontoxic concentrations respectively for 4 days. ELISA was used to detect the HB surface antigen (HBsAg) and HB e antigen (HBeAg) in the supernatant. The relative replication level of HBV DNA was detected by slot blot analysis., Results: MTT method confirmed that the nontoxic concentrations of CsA and FK506 were 0-40.0 microg/ml and 0-400 ng/ml respectively. After the treatment of CsA at the concentration of 1.3, 2.5, and 5.0 microg/ml, in comparison to the control group, the suppression rates of HBsAg expression in the HepG2.2.15 cells were 16.5% +/- 9.4%, 21.5% +/- 8.9%, and 33.1% +/- 5.3% respectively (all P < 0.05); the suppression rates of HBeAg expression in the HepG2.2.15 cells were 7.8% +/- 2.2%, 11.0% +/- 2.3%, and 20.8% +/- 1.5% respectively (all P < 0.05); and the HBV DNA replication levels were 56 +/- 16, 42 +/- 11, and 40 +/- 10 respectively (P > 0.05, P < 0.05, and P > 0.05). However, FK506 at different nontoxic concentrations showed no significant inhibitory effect on the levels of HBsAg, HBeAg, and HBV DNA., Conclusion: CsA dose-dependently inhibits the HBV replication in vitro, and FK506 does not exercise similar effects.

Objective: To explore the effect of the pretreatment with hypertonic saline (HTS) in the hepatic ischemia/reperfusion (I/R) injury., Methods: Twenty-five SD rats were randomly divided into sham operation group, heme oxygenase-1 (HO-1) blocker ZnPP group, I/R group, HTS pretreatment group and ZnPP intervention group (n=5). The rat model of partial hepatic I/R injury was reproduced by isolating the portal venous and hepatic arterial branches to the left and median hepatic lobes, and they were occluded with a microvascular clamp for 30 minutes, followed by reperfusion. In HTS pretreatment group, the rats received 4 ml/kg volume of HTS (7.5%) intravenous 1 hour before the occlusion of the vessels. The rats were sacrificed 6 hours after reperfusion. The levels of alanine aminotransferase (ALT) and tumor necrosis factor-alpha (TNF-alpha) in serum, liver myeloperoxidase (MPO) activity and liver endothelin-1 (ET-1) were determined. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to examine the expression of HO-1 in the liver. The pathological changes in the liver, including hepatic sinusoid, were evaluated in hematoxylin and eosin (HE)-stained sections and transmission electron microscopy (TEM) of liver specimens. The effect of HTS pretreatment was also assessed in the rats pretreated with ZnPP., Results: The levels of serum ALT and TNF-alpha, ET-1 and MPO activity in hepatic tissues increased after hepatic I/R injury (all P<0.01), and expression of HO-1 mRNA and protein were also increased. RT-PCR and Western-blot revealed that the expression of HO-1 in the liver was upgraded significantly, and the ALT level, serum TNF-alpha, liver MPO activity and liver ET-1 were suppressed significantly after I/R injury in the HTS pretreatment group. In this group, there were moderate swelling of hepatocytes and mild neutrophils infiltration in the liver. The hepatic microcirculatory dysfunction was ameliorated. All these findings showed that HTS pretreatment produced the effect of prevention on hepatic I/R injury. However, the adjunctive infusion of ZnPP abrogated the beneficial effects., Conclusion: Pretreatment of HTS has the effect of the prevention of hepatic I/R injury by promotion of the expression of HO-1.

Objective: To explore novel methods of possible donor organ supply and immunologic tolerance induction of organ transplantation., Methods: Whole metanephroi from d14-19 (E14-E19) embryos of pregnant rats were grouped and allografted into the omenta or near remnants of renal vessels of nonimmunosupressed adult rats. At the time of implantation, host rats underwent unilateral nephrectomy. Four weeks after implantation, allografted metanephroi in host rats were removed for gross, biochemical and histopathological examination., Result: Four weeks post-implantation, (1) E19 and E18 metanephroi had enlarged,but were replaced by connective tissues. (2) E17 and E16 metanephroi showed the signs of acute rejection such as hypercellular glomeruli and lymphocyte infiltration in peritubular spaces. E16 grafted metanephroi underwent mild acute rejection of Banff schema, while E17 had moderate or severe acute rejection. When Cyclosporine A was administrated, E17 metanephroi formed mature nephrons and collecting ducts with few lymphocyte infiltration. (3) Metanephroi from E15 and E14 embryos allografted into the omentum or near remnants of renal vessels of uninephrectomized adult rats were enlarged and vascularized, and formed mature tubules and glomeruli. (4) The concentrations of urea nitrogen and creatinine in cyst fluid of E15 and E16 metanephroi were increased 40-fold and 50-fold, which were comparable to those in bladder urine. (5) In contrast, rat metanephroi did not grow or differentiate in rats without host kidney resection., Conclusion: E14 and E15 metanephroi allografted into nonimmunosuppressed adult rats or E17 into cyclosporine-treated hosts undergo growth and differentiation and become vascularized. A variety of factors affect the growth and development of allografted metanephroi, while rejection is the main one.

Objective: To evaluate the effect of ciclosporin A (CsA) on the organogenesis and function of embryonic metanephroi allografted into adult rats., Methods: The whole metanephroi from the 15, 16 and 17 embryonic day-old (E15, E16, E17) embryos of Sprague-Dawley (SD) rat were allografted into the omenta of SD adult rats with their left kidneys resected, which were divided into 2 categories: 3 CsA-treated groups of 10 rats (E15CsASD, E16CsASD, and E17CsASD) and 3 non-CsA-treated groups of 10 rats (E15SD, E16SD, and E17SD). Thirty SD rats without kidney resection were divided into 6 equal groups and underwent allografting embryonic metanephroi in the same manner as mentioned above. The E15 metanephroi of Lewis rats were allografted into the omenta of Thirty adult Brown Norway (BN) rats with one kidney resected. Were divided into 6 equal groups, received allografting of embryonic metanephroi, and injected with CsA of normal saline in the manner as mentioned above (E15CsABN and E15BN). Two to 4 weeks after implantation, the metanephroi allografted in host rats were removed for histopathological examination or anastomosed for renal function measurement 4 weeks later., Results: (1) Four weeks after implantation, the E17SD and E16SD metanephroi showed signs of acute rejection as hypercellular glomeruli and mononuclear cell infiltration in the interstitium. The E16CsASD and E17CsASD metanephroi formed mature nephrons and collecting ducts with few lymphocytic infiltrates. After CsA was discontinued, the E16CsASD and E17CsASD metanephroi were rejected fully within 21 days. (2) 4 weeks after implantation, the E15SD metanephroi were enlarged, became vascularized, and developed mature tubules and glomeruli; however, they were rejected by 100 days after implantation. The E15 Lewis metanephroi were fully rejected within two weeks in the BN adult rats. With CsA administrated, the E15 Lewis metanephroi developed normal mature nephrons and collecting ducts within the adult BN rats. If CsA was discontinued, the E15CsABN metanephroi were rejected. (3) The E15CsASD metanephroi had significantly lower values of wet weight (P = 0.006) and higher values of creatinine clearances (P = 0.007) than the E15SD metanephros transplants, but were identical to those of the E16CsASD metanephroi (P = 0.948, P = 0.840). (4) The metanephroi did not grow or differentiate in the rats without host kidney resection., Conclusions: (1) Cyclosporin A may suppress graft rejection, thus normalizing the growth and function of fetal metanephroi in the omenta of host rats. (2) A variety of factors affect the growth and development of allografted metanephroi, whereas rejection remains the major one.

Objective: To construct the eukaryotic co-expression plasmid of tissue-plasminogen activator (tPA) genes and vascular endothelia growth factor165 (VEGF165) and observe its expression in vascular smooth muscle cells (VSMCs)., Methods: The tPA and VEGF165 genes were cloned from hominal heart tissue with RT-PCR, and then the tPA and VEGF165 genes were cloned into eukaryotic expression plasmid pBudCE4.1 to construct the eukaryotic co-expression plasmid pBudCE4.1/tPA-VEGF165. The pBudCE4.1/tPA-VEGF165 was transfected into VSMCs via lipofection mediation. The expression levels of tPA and VEGF165 mRNAs in the transformed VSMCs were detected by RT-PCR and the expression levels of tPA and VEGF165 proteins were detected by ELISA., Results: The size of RT-PCR product of tPA and VEGF165 genes was 1.9 kb and 576 bp respectively. Restrictive enzyme digestion analysis showed that recombinant co-expression plasmid pBudCE4.1/tPA-VEGF165 had been constructed successfully. The expression of tPA and VEGF165 at mRNA and protein levels in the transformed VSMCs and cells culture supernatant were detected respectively by RT-PCR and ELISA., Conclusion: The recombinant eukaryotic co-expression plasmid pBudCE4.1/tPA-VEGF165 has been successfully constructed. The tPA and VEGF165 are expressed in transformed VSMCs.

Objective: To investigate the effects of immunization with fusions of dendritic cells and H22 cells on tumor-bearing mice and their possible mechanisms., Methods: Fusion cells of DC and H22 cells were prepared with polyethylene glycol (PEG). Expression of MHC and costimulatory molecules by dendritomas were determined by FACs. To study the antitumor immune preventative and therapeutic effects, fusions were subcutaneously injected into tumor-bearing mice. The cytotoxic T lymphocyte (CTL) activity was determined by LDH method, the expression of TNF-a and IFN-g in tumors were assayed by RT-PCR., Results: The data showed that the hybridomas of DC and H22 cells acquired both DC and H22 cell phenotypes. Immunization of BALB/C mice with DC/H22 fusions induced potent CTL activity (mean CTL activity=0.624+/-0.024, compared with DC + H22, DC, H22 groups, F = 65.46) and a protective immunity against a high dose of H22 tumor challenge. After treatment with hybridomas, the survival time of tumor-bearing mice was greatly extended (x2=18.45). The expression levels of TNF-a and IFN-g mRNA were remarkably increased (TNF-a, F = 47.84; IFN-g, F = 37.23)., Conclusions: The hybridomas of DC and H22 cells could induce effective antitumor immune responses and may have a useful potential in prevention and management of the recurrences and metastases of HCC.

Objective: To investigate the relation between the activity of nuclear factor kappaB (NF-kappaB) and the expression of interferon (IFN)-gamma gene transcription with or without ciclosporin treatment after liver transplantation., Methods: Orthotopic liver transplantation was performed in this study. Group I: syngeneic control (Wistar-to-Wistar); Group II: acute rejection (SD-to-Wistar); Group III: acute rejection treated with ciclosporin by intramuscular route (SD-to-Wistar + ciclosporin). Electrophoretic mobility shift assay and reverse transcriptase polymerase chain reaction were used to analyze NF-kappaB activity of splenocytes and IFN-gamma gene transcription expression of grafted liver with or without ciclosporin treatment after liver transplantation. Histopathological examination was also used in this study., Results: Low NF-kappaB activity was only detected at day 5 and day 7 in Wistar-to-Wistar group after transplantation, meanwhile low IFN-gamma mRNA expression was detected at any time in this group. In contrast, high NF-kappaB activity was detected in SD-to-Wistar group and high level IFN-gammamRNA expression was detected at all time points in this group. The activity of NF-kappaB and IFN-gammamRNA expression were significantly inhibited in SD-to-Wistar + ciclosporin group which was significantly lower than that of SD-to-Wistar group (P < 0.001). A good correlation was found between activity of NF-kappaB and IFN-gamma mRNA expression in this study (r=0.815, P <0.01)., Conclusions: The change of expression IFN-gamma mRNA is at least partially due to the activity change of NF-kappaB after orthotopic liver transplantation. CsA down-regulates NF-kappaB activity and further inhibit IFN-gamma gene transcription.

A tissue culture system for embryogenic callus (EC) induction and plant regeneration of common bermudagrass using mature caryopsis (embryos) as explants was developed. The results showed that embryogenic calli could be induced from caryopsis with high frequency, in MS medium with 2,4-D 2.0-6.0 mg/L, and the best concentration of 2,4-D was 4.0 mg/L. The best method for maintaining EC and tissue differentiation was to subculture EC in MS+2,4-D 4.0 mg/L 1-2 times, follwed by subculturing in 1/2 MS+2,4-D 2.0 mg/L for 1-2 times, then to transfer EC to 1/2 MS without hormone for a 10-d-preregeneration in light, followed by transferring to MS+6-BA 3.0 mg/L for regeneration, with regeneration frequency 31.7%. Morphological and micro-structural differences between EC and non-embryogenic callus (NEC) were observed by electron microscope. Ultrastructrual characteristics of the EC cells are described in this paper.

Objective: To investigate the effect of combined CsA and FK506 with 5-FU on hepatocellular carcinoma rats., Methods: A syngeneic rat model of hepatocellular carcinoma was used. Control group (A) underwent 4 ml 5% GS. Treatment group was divided into 3 groups namely, group B: only 5-FU and 5% GS; group C: 5-FU, CsA and 5% GS; group D: 5-FU, FK506 and 5%GS. Cell cycle, apoptosis, necrosis and mitochondrial transmembrane potential were measured by flow cytometry, laser scanning confocal microscopy, and electron transmission microscopy. Statistical analysis was performed by SPSS 10.0 for Windows software. Statistical comparisons were made with ANOVA followed by Dunnett's T3 or LSD test., Results: Compared to the control group, the percentage of apoptotic cells including trifle necrotic cells was significantly higher, and among the treatment group, group D was the highest, and group C was higher than group B. In the treatment group, cell cycle of hepatoma cells was mainly arrested at S phase, but in group D, G0/G1 phase cells were significantly decreased and S phase cells significantly increased. Compared to the control group, mitochondrial transmembrane potential was significantly decreased in the treatment group, among with, group B was the lowest, group C was higher than group D. Morphological changes demonstrated by electron microscopy included dispersed nuclear chromatin, loss of nucleoli, membrane bleeding, cell shrinkage, typical apoptotic bodies and marked swelling of mitochondria in the treatment group. In the control group, however, they were characterized by normal cell ultrastructure., Conclusions: The present study reveals that 5-FU combined with CsA or FK506 demonstrated a synergistic effect on hepatocellular carcinoma rats. For FK506, the powerful mutual effect is related to the increase of tumor cell's quantity in S phase. Both CsA and FK506 can provide protection on mitochondrial transmembrane potential reduction against hepatoma cells damage from 5-FU.

OBJECTIVE: To evaluate the relatinship between the expression of P51, P73 and the oncogenesis and development of human gastric carcinoma. METHODS: The expression of P73 mRNA were detected both in 32 human gastric carcinoma tissues and adjacent normal gastric tissues by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Overexpressions of P73 mRNA were found in 17/32 gastric carcinoma tissues,in 2/32 adjacent normal gastric tissues.The positive expression rate of P73 mRNA in gastric carcinooma tissues was significantly higher than in adjacent normal gastric tissues( P<0.01). However, a significant correlation was found between the positive expression rate of P73 mRNA in gastric carcinoma tissues and the TNM staging(P<0.05). THe low expressions of P51A mRNA and P51B were found in all gastric carcinoma tissues and adjacent normal gastric tissues. The expression of P51A in gastric carcinoma tissues were much higher than adjacent normal gastric tissues (P<0.05). The expression of P51B is no significant correlation was observed between gastirc carcinoma tissues and adjacent normal gastric tissues. CONCLUSION: The results suggest that there is an overexpression odf P73 and P51A mRNA in gastric cancer tissues, and their expressions is relationship with oncogenesis and developnment of gastric carcinoma.

The effect of IL-3 and hydroxyurea on human erythroleukemia cell line (K562 cells) was demonstrated by using the electro-microscopy and flow cytometry. Our data showed that neither IL-3 nor hydroxyurea could induce the apoptosis of K562 cells alone. However, the IL-3 and hydroxyurea could induce the apoptosis of K562 cells cooperatively. Analysis with flow cytometry showed that the percentage of apoptotic cells was about 31.90% after K562 cells were induced by IL-3 and hydroxyurea cooperatively for 5 days, and the sub-G1 peak (apoptotic peak) was detected in the induced K562 cells. Meanwhile, the percentage of S-phase in the IL-3 and hydroxyurea induced K562 cells was increased, and the proliferation of the induced K562 cells was inhibited significantly. Furthermore, the IL-3 and hydroxyurea induced K562 cells showed chromatin condensation with regular crescents at the nuclear edges and apoptotic bodies. It suggested that IL-3 could enhance the sensitivity of K562 cells to hydroxyurea and the apoptosis of K562 cells could be induced by IL-3 and hydroxyurea cooperatively.

A yeast (Saccharomyces cerevisiae) secretory expression vector containing PRO3 gene was constructed (pCBy310). beta HCG(Human choriogonadotropin beta subunit)-cDNA was inserted into pCBy310 to form a recombinant plasmid pCBy314. Since yeast proline auxotroph will not survive in rich medium (YPD), YPD could be used as a selection pressure, and pCBy314 could be maintained mitotically stable in transformants of yeast Pro3- auxotroph (MB299-7A) in rich medium. At an improved, yet not optimized cultural condition, the expression of beta HCG in culture medium was 650 micrograms/L. Our results showed not only that YPD could be used as a selection medium, but also that yeast grew better in YPD so as to increase the gene expression product, and that the component of YPD was simple and cheap. Our data suggested that PRO genes might be used widely in constructing vectors to clone and express foreign genes in yeast so that rich medium can be used as a selection pressure for direct selection.