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Fatty acid synthase (FASN) is known as a crucial enzyme of cellular de novo fatty acid synthesis in mammary gland which has been proved as the main source of short and medium-chain fatty acids of milk. However, the regulatory role of FASN in goat-specific milk fatty acids composition remains unclear. We cloned and analyzed the full-length of FASN gene from the mammary gland of Capra hircus (Xinong Saanen dairy goat) (DQ 915966). Comparative gene expression analysis suggested that FASN is predominantly expressed in fat, small intestine and mammary gland tissues, and expresses higher level at lactation period. Inhibition of FASN activity by different concentrations (0, 5, 15, 25 and 35 μM) of orlistat, a natural inhibitor of FASN, resulted in decreased expression of acetyl-CoA carboxylase α (ACCα), lipoprotein lipase and heart-type fatty acid binding protein (H-FABP) in a concentration-dependent manner in goat mammary gland epithelial cells (GMEC). Similar results were also obtained by silencing of FASN. Additionally, reduction of FASN expression also led to apparent decline of the relative content of decanoic acid (C10:0) and lauric acid (C12:0) in GMEC. Our study provides a direct evidence for inhibition of FASN reduces cellular medium-chain fatty acids synthesis in GMEC. [ABSTRACT FROM PUBLISHER]

Myc oncoproteins are commonly upregulated in human cancers of different organ origins, stabilized by Aurora A, degraded through ubiquitin-proteasome pathway-mediated proteolysis, and exert oncogenic effects by modulating gene and protein expression. Histone deacetylases are emerging as targets for cancer therapy. Here we demonstrated that the class III histone deacetylase SIRT2 was upregulated by N-Myc in neuroblastoma cells and by c-Myc in pancreatic cancer cells, and that SIRT2 enhanced N-Myc and c-Myc protein stability and promoted cancer cell proliferation. Affymetrix gene array studies revealed that the gene most significantly repressed by SIRT2 was the ubiquitin-protein ligase NEDD4. Consistent with this finding, SIRT2 repressed NEDD4 gene expression by directly binding to the NEDD4 gene core promoter and deacetylating histone H4 lysine 16. Importantly, NEDD4 directly bound to Myc oncoproteins and targeted Myc oncoproteins for ubiquitination and degradation, and small-molecule SIRT2 inhibitors reactivated NEDD4 gene expression, reduced N-Myc and c-Myc protein expression, and suppressed neuroblastoma and pancreatic cancer cell proliferation. Additionally, SIRT2 upregulated and small-molecule SIRT2 inhibitors decreased Aurora A expression. Our data reveal a novel pathway critical for Myc oncoprotein stability, and provide important evidences for potential application of SIRT2 inhibitors for the prevention and therapy of Myc-induced malignancies. [ABSTRACT FROM AUTHOR]

In the swine industry, manure and odor treatment are one of the important issues. The purpose was analyze the effects of Lactobacillus kefiranofaciens M1, Bacillus subtilis S14 and B. amyloliquefaciens S20 on enhancing the growth performance, reducing fecal noxious gas emission, and increasing carcass quality of grow-finishing pig. The possible mechanism involving the modifying intestinal microbiota was also investigated. Thus, in the present study, different combinations of probiotics were fed to grow-finishing Pigs. The swine growth performance, fecal odor, and blood biochemistry were monitored. After slaughter, the carcass characteristics were assessed. The possible mechanisms involving in changes of gut microbiota were also investigated. In addition, full length 16S rRNA analysis was used to involved changes in the gut microbiota. Results showed that administration of the selected probiotics could increase the growth performance, reduce the fecal odor, and improve carcass quality. Further, microbiota analysis demonstrated that the taxa with significantly differences in abundances among groups were paralleled with linear discriminant analysis (LDA) effect size (LEfSe) finding, which were identified as the critical gastrointestinal bacterial biomarkers. Expected that the potential probiotic strains selected could be used as feed supplements to provide a possible solution for the difficulties faced by the swine industry. [ABSTRACT FROM AUTHOR]

The role of fatty acid synthase (FASN) on de novo fatty acid synthesis has been well established. In monogastrics, unlike acetyl-coenzyme A carboxylase, FASN is primarily controlled at the transcriptional level. However, no data exist on ruminant mammary cells evaluating effects of FASN knockdown on mRNA expression of lipogenic genes. Inhibition of FASN in mammary cells by C75-mediated interference, a synthetic inhibitor of FASN activity, and short hairpin RNA-mediated interference markedly reduced cellular triglyceride content at least in part by decreasing the expression of genes related to triglyceride synthesis (GPAT, AGPAT6, and DGAT2) and enhancing the expression of lipolysis-related genes (ATGL and HSL). Consistent with the markedly lower expression of genes related to lipid droplet formation and secretion (TIP47, ADFP, BTN1A1, and XDH), cellular lipid droplets also were reduced sharply after incubation with C75 or adenovirus-short-hairpin-RNA. The results underscored the essential role of FASN in the overall process of milk-fat formation in goat mammary epithelial cells. [ABSTRACT FROM AUTHOR]

Milk fat originates from the secretion of cytosolic lipid droplets (CLD) synthesized within mammary epithelial cells. Adipocyte differentiation-related protein (ADRP; gene symbol PLIN2) is a CLD-binding protein that is crucial for synthesis of mature CLD. Our hypothesis was that ADRP regulates CLD production and metabolism in goat mammary epithelial cells (GMEC) and thus plays a role in determining milk fat content. To understand the role of ADRP in ruminant milk fat metabolism, ADRP (PLIN2) was overexpressed or knocked down in GMEC using an adenovirus system. Immunocytochemical staining revealed that ADRP localized to the surface of CLD. Supplementation with oleic acid (OA) enhanced its colocalization with CLD surface and enhanced lipid accumulation. Overexpres-sion of ADRP increased lipid accumulation and the concentration of triacylglycerol in GMEC. In contrast, morphological examination revealed that knockdown of ADRP decreased lipid accumulation even when OA was supplemented. This response was confirmed by the reduction in mass of cellular TG when ADRP was knocked down. The fact that knockdown of ADRP did not completely eliminate lipid accumulation at a morphological level in GMEC without OA suggests that some other compensatory factors may also aid in the process of CLD formation. The ADRP reversed the decrease of CLD accumulation induced by adipose triglyc-eride lipase. This is highly suggestive of ADRP promoting triacylglycerol stability within CLD by preventing access to adipose triglyceride lipase. Collectively, these data provide direct in vitro evidence that ADRP plays a key role in CLD formation and stability in GMEC. [ABSTRACT FROM AUTHOR]

In nonruminants, the alternative splicing of peroxisome proliferator-activated receptor γ (PPARG) generates PPARG1 and PPARG2 isoforms. Although transcriptional control differences between isoforms have been reported in human adipose tissue, their roles in ruminant mammary cells are not well known. To assess which of these isoforms is more closely associated with the regulation of mammary lipogenic pathways, their tissue distribution was analyzed and the expression of key genes regulating lipogenic gene networks was measured after overexpression of the 2 isoforms in goat mammary epithelial cells (GMEC). The expression of PPARG2 was markedly greater in adipose tissue, whereas PPARG1 is the main isoform in goat mammary tissue (ratio of PPARG1:PPARG2 was close to 37:1). As was reported in previous work, PPARG1 upregulated the transcription regulators SREBF1 and PPARG and the lipogenic genes FASN, ACACA, and SCD. Along with a tendency for greater expression of AGPAT6, DGAT1, and PLIN2, these data suggest that PPARG1 is the isoform controlling lipogenesis in mammary cells. Addition of the PPARG ligand rosiglitazone (ROSI) to GMEC overexpressing both isoforms upregulated the expression of LPL and CD36, which help control uptake of long-chain fatty acids into mammary cells. Other responses to ROSI addition to GMEC overexpressing PPARG1 and PPARG2 included upregulation of AGPAT6, DGAT1, INSIG1, SREBF1, and NR1H3. Although the data suggest that both PPARG1 and PPARG2 could affect mammary lipogenesis via control of gene expression when stimulated (e.g., by ROSI), the fact that PPARG1 is more abundant in mammary tissue and that its overexpression alone upregulated key lipogenic gene networks suggest that it is the more important isoform in goat mammary cells. [ABSTRACT FROM AUTHOR]

Objective: To investigate the regulative effects of Streptococcus mutans (Sm) antisense vicK RNA (ASvicK) on the multi-species biofilm formed by three common oral streptococci (Sm, Streptococcus sanguinis , and Streptococcus gordonii ) (Sm+Ss+Sg). Methods: ASvicK over-expression strain was constructed by using a recombinant plasmid, and three-species biofilm UA159+Ss+Sg and ASvicK+Ss+Sg were cultured. The phenotypes of biofilms were detected by scanning electron microscopy (SEM). Crystal violet (CV) assay was used to detect biofilm biomass. Lactate kit and anthrone-sulfuric acid colorimetric assay were used to determine the abilities of lactic acid and exopolysaccharides production, respectively. The proportions of three-species and expression levels of the cariogenic-related genes in biofilms were detected by TaqMan fluorescence quantitative PCR and real-time fluorescence quantitative PCR. A biofilm demineralization model of human enamel slabs was further constructed, and the hardness of enamel surface was detected. Results: Compared to UA159+Ss+Sg, over-expression of ASvicK could inhibit biofilm formation and lactic acid production in ASvicK+Ss+Sg biofilm significantly decreased by 78.93% ( P< 0.001) and 62.23% ( P< 0.001), respectively. With ASvicK over-expression, the amounts of water-insoluble and-soluble glucoses in ASvicK+Ss+Sg biofilm were reduced respectively by 39.13% ( P< 0.001) and 68.00% ( P< 0.001). Compared to the UA159+Ss+Sg Group, the proportion of Sm, the cariogenic bacteria, showed 33.00% reduction ( P< 0.01) in Sm+Ss+Sg biofilm, and the gene expressions of cariogenic-relative genes vicK/X, gtfB/C/D, and ftf significantly decreased ( P< 0.05). The micro-hardness value of enamel slabs after demineralization by ASvicK+Ss+Sg biofilm was significantly increased to 183.84% ( P< 0.001). Conclusions: ASvicK over-expression could reduce the Sm proportion and weaken the cariogenicity of oral Streptococcus biofilm, thereby possibly slowing down the progression of caries.

Objective: To establish a method for the determination of chlorothalonil in air by filtration membrane sampling, solvent elution and gas chromatography. Methods: PTFE filter was used for sampling, eluted with dichloromethane, separated by DB-5 capillary column, and determined by FID. Results: The standard curve was used for quantitative detection, and the correlation of chlorothalonil in the range of 15 μg/ml-300 μg/ml, R (2)=0.9999. The detection limit of this method was 1.70 μg/ml and the lower quantitative limit was 5.70 μg/ml. The minimum detected concentration was 0.045 mg/m(3) (75L air sample was collected) . The recovery rate was 90.14%-91.81%. The precision of the same batch was 1.5%-1.8%, and that of different batches was 2.3%-3.8%. The sampling efficiency can reach above 95%; The samples can be stored for 14 days at room temperature. Conclusion: The results show that the method of filtration membrane sampling-solvent elude-gas chromatography is suitable for the determination of chlorothalonil in the air.

Objective: To investigate the correlation between tooth movement and profile change in patients with class Ⅱ division 1 malocclusion. Methods: Pre- and post-treatment lateral cephalograms of 42 patients [10 males and 32 females, (23.8±6.3) years old, mean treatment time: 1.9 years] with class Ⅱ division 1 malocclusion were collected in Department of Oral & Cranio-Maxillofacial Surgery, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from June 2012 to November 2017. The patients were treated with extraction of four first premolars or two maxillary first premolars. Cephalometric analysis was carried out before and after treatment. Thirty parameters were measured. The changes of soft and hard tissue after orthodontic treatment and their correlations were analyzed using bivariate linear regression. Related factors affecting the upper and lower lip, nasolabial angle (NLA) and mentolabial angle (MLA) were analyzed according to the standardized regression coefficient ( Beta ). Results: Among all the 30 parameters, 18 parameters were statistically different before and after treatment. After treatment, upper central incisor sagittal distance [(63.87±7.14) mm] and upper lip sagittal distance [(77.73±7.60) mm] were significantly decreased ( P <0.05). The changes in 14 parameters after treatment showed linear relationship including strong positive correlation between upper lip sagittal retraction and upper central incisor sagittal retraction ( r =0.649, P <0.01). There were moderate positive correlations between upper lip and upper central incisor vertical movement ( r =0.544, P <0.01). While the sagittal change of gnathion and the Y-axis angle showed moderate negative correlations ( r =0.537, P <0.01). The stepwise multiple linear regression showed that the retraction of upper lip process was correlated with the retraction of upper central incisor, the increase of occlusal plane angle and the increase of upper central incisor angle, which was most correlated with the retraction of upper central incisor ( Beta =0.79). The downward displacement of upper lip process was correlated with the downward displacement of upper incisor, the decrease of upper central incisor angle, the decrease of the distance between maxillary first molar and palatal plane, and the increase of occlusal plane angle, which was more correlated with the downward displacement of upper incisor and the increase of occlusal plane angle ( Beta =0.59). The downward displacement of lower lip process was correlated with the downward displacement of upper incisor and lower incisor, which was more correlated with the upper incisor ( Beta =0.36). Conclusions: The relationship among nose, lips and chin was more coordinated. Incisor retraction had significant influence on lip prominence, and the lower lip position was highly related to the movement of upper incisor in sagittal and vertical dimension after orthodontic treatment in patients with class Ⅱ division 1 malocclusion. However, tooth movement had limited impact on the chin position.

Objective: To analyze and summarize the prevalence of diabetic retinopathy (DR) in China, and provide scientific evidence for its prevention and intervention. Methods: Literature search of PubMed, Web of Science, Embase, Wanfangdata, Vip and CNKI between 1990 and 2020 was performed to retrieve epidemiological studies of DR in China. The total prevalence of DR was analyzed by performing a meta-analysis, and the prevalences of DR in different regions and age groups were also evaluated by subgroup analyses. Results: A total of 40 studies were enrolled. The total sample size of the general population was 282 620, and the total number of diabetic patients was 47 022. The prevalence of DR in the general population and diabetic patients was 1.7% (95 %CI : 1.4%-2.0%) and 22.4% (95% CI: 18.8%-26.1%), respectively. The highest prevalence of DR was found in patients aged 50-59 years (22.1%). Among the diabetic population, the prevalence of DR was high in North (27.7%) and Northeast China (23.7%), but the East China had the largest estimated number of DR patients (4 971 000). Moreover, the prevalence of DR in rural areas (34.0%) was higher than that in urban areas (18.7%). Conclusions: The prevalence of DR is high in Chinese diabetic population, and there are differences among regions and age subgroups. The results of the current meta-analysis emphasize the necessity of DR screening for diabetic population, especially in rural areas.

Objectives: To evaluate the safety and efficacy of chidamide to reverse HIV-1 latency in vivo and to compare the effects of four clinically tested histone deacetylase (HDAC) inhibitors on non-histone proteins in vitro., Methods: Participants received chidamide orally at 10 mg twice weekly for 4 weeks while maintaining baseline antiretroviral therapy. The primary outcome was plasma viral rebound during chidamide dosing and the secondary outcomes were safety, pharmacokinetic and pharmacodynamic profiles, changes in cell-associated HIV-1 RNA and HIV-1 DNA, and immune parameters. Western blotting was used to compare the in vitro effects of the four HDAC inhibitors on HSP90, NF-κB and AP-1., Results: Seven aviraemic participants completed eight oral doses of chidamide, and only grade 1 adverse events were observed. Cyclic increases in histone acetylation were also detected. All participants showed robust and repeated plasma viral rebound (peak viraemia 147-3850 copies/mL), as well as increased cell-associated HIV-1 RNA, during chidamide treatment. Furthermore, we identified an enhanced HIV-1-specific cellular immune response and a modest 37.7% (95% CI: 12.7-62.8%, P = 0.028) reduction in cell-associated HIV-1 DNA. Compared with the other three HDAC inhibitors, chidamide had minimal cytotoxicity in vitro at clinically relevant concentrations and showed mechanistically superior effects on non-histone proteins, including HSP90, NF-κB and AP-1., Conclusions: Chidamide safely and vigorously disrupts HIV-1 latency in vivo, which makes it a promising latency-reversing agent., (© 2020 British HIV Association.)

Objective: To understand the clinical characteristics, change of liver function, influencing factors and prognosis in hospitalized patients with coronavirus disease-19 (COVID-19) combined with liver injury. Methods: The general conditions, biochemical indicators of liver, blood clotting mechanism, routine blood test, UGT1A1 * 28 gene polymorphism and other data of 40 cases with COVID-19 admitted to the isolation ward of Tangdu Hospital were retrospectively analyzed. The clinical characteristics, influencing factors and prognosis of liver injury in patients with liver injury group and those with normal liver function group were compared. The mean of two samples in univariate analysis was compared by t-test and analysis of variance. The counting data was measured by χ(2) tests. The non-normal distribution measurement data were described by the median, and the non-parametric test was used. Statistically significant influencing factors were used as the independent variables in univariate analysis. Multiple logistic regression analysis was used to analyze the main influencing factors of liver injury. Results: Of the 40 cases, 25 were male (62.5%) and 15 were female (37.5%), aged 22 to 83 (53.87 ± 15.84) years. Liver injury was occurred in 22 cases (55%) during the course of the disease. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) level was initially increased (4.4 to 3.5 times of the normal value) along with decrease of albumin in the second week, and the difference was statistically significant ( P < 0.001). Ten cases (43.5%) had highest abnormal total blood bilirubin (54.1 μmol/ L). There was no correlation between the increase in transaminase and the increase in total blood bilirubin ( R = -0.006, P = 0.972). Three cases had prothrombin activity (PTA) of ≤50%, 10 cases had elevated FDP, and 13 cases had elevated D-dimer, all of whom were severe or critically ill. Liver function injury was more likely to occur in patients who used many types of drugs and large amounts of hormones ( P = 0.002, P = 0.031), and there was no correlation with the TA6TA7 mutation in the UGT1A1 * 28 gene locus. Multiple regression analysis showed that the occurrence of liver injury was only related to critical illness. The liver function of all patients had recovered within one week after conventional liver protection treatment. Conclusion: COVID-19 combined with liver function injury may be due to the slight elevation of transaminase, mostly around the second week of the disease course. Severe patients have a higher proportion of liver injury, and critical type is an independent risk factor for liver injury.

Objective: Hepatitis B surface antigen (HBsAg) loss is seldom achieved with nucleos(t)ide analog (NA) therapy in chronic hepatitis B patients but may be enhanced by switching to finite pegylated-interferon (Peg-IFN) alfa-2a. We assessed HBsAg loss with 48- and 96-week Peg-IFN alfa-2a in chronic hepatitis B patients with partial response to a previous NA. Methods: Hepatitis B e antigen (HBeAg)-positive patients who achieved HBeAg loss and hepatitis B virus DNA < 200 IU/mL with previous adefovir, lamivudine or entecavir treatment were randomized 1:1 to receive Peg-IFN alfa-2a for 48 ( n = 153) or 96 weeks ( n = 150). The primary endpoint of this study was HBsAg loss at end of treatment. The ClinicalTrials.gov identifier is NCT01464281. Results: At the end of 48 and 96 weeks' treatment, 14.4% (22/153) and 20.7% (31/150) of patients, respectively, who switched from NA to Peg-IFN alfa-2a cleared HBsAg. Rates were similar irrespective of prior NA or baseline HBeAg seroconversion. Among those who cleared HBsAg by the end of 48 and 96 weeks' treatment, 77.8% (14/18) and 71.4% (20/28), respectively, sustained HBsAg loss for a further 48 weeks. Baseline HBsAg < 1 500 IU/mL and week 24 HBsAg < 200 IU/mL were associated with the highest rates of HBsAg loss at the end of both 48- and 96-week treatment (51.4% and 58.7%, respectively). Importantly, extending treatment from 48 to 96 weeks enabled 48.3% (14/29) more patients to achieve HBsAg loss. Conclusion: Patients on long-term NA who are unlikely to meet therapeutic goals can achieve high rates of HBsAg loss by switching to Peg-IFN alfa-2a. HBsAg loss rates may be improved for some patients by extending treatment from 48 to 96 weeks, although the differences in our study cohort were not statistically significant. Baseline and on-treatment HBsAg may predict HBsAg loss with Peg-IFN alfa-2a.

Objective: To offer the objective evidences for the diagnosis by analyzing anxiety and depression among patients with sudden deafness and vertigo and their possible causes. Method: Eighty patients with sudden deafness and vertigo of single ear as the observation group; 80 patients with sudden deafness without vertigo and 60 healthy people as control group.Hamilton anxiety scale (HAMA), Hamilton depression scale (HAMD) are used for quantitative analysis and compare the results of each group score. Result: ①The HAMA score of the deafness group with vertigo was significantly higher than that of the group without vertigo and the healthy group ( P <0.01). The HAMD score of the deafness group with vertigo was significantly higher than that of the group without vertigo and the healthy group ( P <0.01). ②There was correlation between HAMA and sex, age, duration of dizziness, tinnitus, DHI and efficacy grading ( P <0.05). There was no correlation between HAMA and the mean hearing threshold, urban and rural distribution, ear fullness ( P > 0.05). There was a correlation between HAMD and sex, age, tinnitus, DHI and efficacy grading ( P <0.05). There was no correlation between HAMD and the mean hearing threshold, duration of dizziness, urban and rural distribution, ear fullness ( P >0.05). Conclusion: There are anxiety and depression in patients with deafness and vertigo. It is mainly related to sex, age, dizziness duration, tinnitus, DHI and grade of efficacy. It is necessary to pay attention to the mental state of patients in clinical treatment. Psychological counseling or treatment can be used to improve clinical 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 Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval ( CI ) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion: In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.

Objective: To investigate the efficacy and safety of the new investigational drug pegylated interferon α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 µg/week) combined with ribavirin in the treatment of patients with genotype 1/6 chronic hepatitis C (CHC), with standard-dose Peg-IFN-α-2a combined with ribavirin as a positive control. Methods: A multicenter, randomized, open-label, and positive-controlled phase III clinical trial was performed. Eligible patients with genotype 1/6 CHC were screened out and randomly divided into Peg-IFN-α-2b(Y shape, 40kD) group and Peg-IFN-α-2a group at a ratio of 2:1. The patients in both groups were given oral ribavirin for 48 weeks in addition and then followed up for 24 weeks after drug withdrawal. Abbott Real Time HCV Genotype II was used to determine HCV genotype, and Cobas TaqMan quantitative real-time PCR was used to measure HCV RNA level at 0, 4, 12, 24, 48, and 72 weeks. Adverse events were recorded in detail. The primary efficacy endpoint was sustained virological response (SVR), and a non-inferiority test was also performed. Results: A total of 561 patients with genotype 1/6 CHC were enrolled, among whom 529 received treatment; 90.9% of these patients had genotype 1 CHC. The data of the full analysis set showed that SVR rate was 69.80% (95% CI 65.00%-74.60%) in the trial group and 74.16% (95% CI 67.73%-80.59%) in the control group ( P = 0.297 0). The data of the per protocol set (PPS) showed that SVR rate was 80.63% (95% CI 76.04%-85.23%) in the trial group and 81.33% (95% CI 75.10%-87.57%) in the control group ( P = 0.849 8), and the 95% CI of rate difference conformed to the non-inferiority standard. The analysis of the PPS population showed that of all subjects, 47.9% achieved rapid virologic response, with a positive predictive value of 93.8%. The incidence rate of adverse events was 96.30% in the trial group and 94.94% in the control group, and the incidence rate of serious adverse events was 5.13% in the trail group and 5.06% in the control group. Conclusion: In the regimen of Peg-IFN-α combined with ribavirin for the treatment of genotype 1/6 CHC, the new investigational drug Peg-IFN-α-2b(Y shape, 40 kD) has comparable clinical effect and safety to the control drug Peg-IFN-α-2a.

The aim of the current study was to explore the effect of the ketamine on the immune function and cognitive function in young rats. The young rats (40) rats were randomly divided into two groups where each group contains 20 rats, such as Group I: normal control; Group II: Ketamine treated group. All group rats received the intravenous injection of treatment for three times and the hippocampal neuronal apoptosis and the immune parameters such as IL-2, IL-4 and IL-10 and whole brain IL-1β level were estimated. The cognitive ability effect of the young rats was also tested using the Morris water maze test. In Morris water maze test, it has been found, as the time increases, the latency of the control group and ketamine treated groups rats were gradually decreased, with a significant difference. The latency rate of the control group was unchanged significantly (P<0.05), but after 3 days, the latency has been decreased significantly. The hippocampal neuronal apoptosis of the control group and ketamine treated group rats were found to be 13.5×5.8 % and (2.1×1.4) %, respectively. The level of the serum IL-4 and IL-10 were also found significantly (P<0.05) higher in the ketamine group as compared to the control group rats. The level of the IL-2 was found to be almost similar in both normal control and ketamine group rats. Markedly, the level of the whole brain IL-1β was found to be significantly higher in the ketamine treated group in comparison to the control group rats. On the basis of the above fact, it has been conclude that the ketamine might be able to inhibit the cognitive function as well as immune function.

The role of p44/42 mitogen-activated protein kinase (MAPK), p38, and c-Jun NH(2)-terminal kinase (JNK) in tumor necrosis factor (TNF)-alpha-induced cyclooxygenase (COX)-2 expression was studied in NCI-H292 epithelial cells. TNF-alpha-mediated COX-2 expression and COX-2 promoter activity were inhibited by the MAPK kinase inhibitor PD98059 or the p38 inhibitor SB203580. Treatment of cells for 10 min with TNF-alpha resulted in activation of p44/42 MAPK, p38, and JNK. C2-ceramide (a cell-permeable ceramide analog), bacterial neutral sphingomyelinase (Smase; an enzyme that degrades sphingomyelin to ceramide), and N-oleoylethanolamine (a ceramidase inhibitor) all induced activation of MAPKs, COX-2 expression, nuclear factor (NF)-kappaB DNA-protein binding, and COX-2 promoter activity. The inactive analog, dihydro-C2-ceramide, had no effect. SMase- or C2-ceramide-induced COX-2 expression and COX-2 promoter activity were also inhibited by PD98059 or SB203580. Glutathione, a neutral SMase inhibitor, attenuated TNF-alpha- or SMase-induced activation of MAPKs, COX-2 expression, and COX-2 promoter activity. TNF-alpha- or C2-ceramide-induced COX-2 promoter activity was inhibited by the dominant negative mutant of extracellular signal-regulated kinase 2, p38, JNK, IkappaB kinase (IKK)1, or IKK2. IKK activity was stimulated by either TNF-alpha or C2-ceramide, and these effects were inhibited by PD98059 or SB203580. All these results suggest that, in NCI-H292 epithelial cells, activation of MAPKs by ceramide contributes to the TNF-alpha signaling that occurs downstream of neutral SMase activation and results in the stimulation of IKK1/2, and NF-kappaB in the COX-2 promoter, followed by initiation of COX-2 expression.

TNF-alpha induced a dose- and time-dependent increase in cyclooxygenase-2 (COX-2) expression and PGE2 formation in human NCI-H292 epithelial cells. Immunofluorescence staining demonstrated that COX-2 was expressed in cytosol and nuclear envelope. Tyrosine kinase inhibitors (genistein or herbimycin) or phosphoinositide-specific phospholipase C inhibitor (U73122) blocked TNF-alpha-induced COX-2 expression. TNF-alpha also stimulated phosphatidylinositol hydrolysis and protein kinase C (PKC) activity, and both were abolished by genistein or U73122. The PKC inhibitor, staurosporine, also inhibited TNF-alpha-induced response. The 12-O-tetradecanoylphorbol 13-acetate (TPA), a PKC activator, also stimulated COX-2 expression, this effect being inhibited by genistein or herbimycin. NF-kappaB DNA-protein binding and COX-2 promoter activity were enhanced by TNF-alpha, and these effects were inhibited by genistein, U73122, staurosporine, or pyrolidine dithiocarbamate. TPA stimulated both NF-kappaB DNA-protein binding and COX-2 promoter activity, these effects being inhibited by genistein, herbimycin, or pyrolidine dithiocarbamate. The TNF-alpha-induced, but not the TPA-induced, COX-2 promoter activity was inhibited by phospholipase C-gamma2 mutants, and the COX-2 promoter activity induced by either agent was attenuated by dominant-negative mutants of PKC-alpha, NF-kappaB-inducing kinase, or I-kappaB (inhibitory protein that dissociates from NF-kappaB) kinase (IKK)1 or 2. IKK activity was stimulated by both TNF-alpha and TPA, and these effects were inhibited by staurosporine or herbimycin. These results suggest that, in NCI-H292 epithelial cells, TNF-alpha might activate phospholipase C-gamma2 via an upstream tyrosine kinase to induce activation of PKC-alpha and protein tyrosine kinase, resulting in the activation of NF-kappaB-inducing kinase and IKK1/2, and NF-kappaB in the COX-2 promoter, then initiation of COX-2 expression and PGE2 release.

The interaction of apolipoprotein H (Apo H) with lipid membrane has been considered to be a basic mechanism for the biological function of the protein. Previous reports have demonstrated that Apo H can interact only with membranes containing anionic phospholipids. Here we study the membrane-induced conformational change of Apo H by CD spectroscopy with two different model systems: anionic-phospholipid-containing liposomes [such as 1, 2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) and cardiolipin], and the water/methanol mixtures at moderately low pH, which mimic the micro-physicochemical environment near the membrane surface. It is found that Apo H undergoes a remarkable conformational change on interaction with liposomes containing anionic phospholipid. To interact with liposomes containing DMPG, there is a 6.8% increase in alpha-helix in the secondary structures; in liposomes containing cardiolipin, however, there is a 12.6% increase in alpha-helix and a 9% decrease in beta-sheet. The similar conformation change in Apo H can be induced by treatment with an appropriate mixture of water/methanol. The results indicate that the association of Apo H with membrane is correlated with a certain conformational change in the secondary structure of the protein.

The interaction of rabbit C-reactive protein (rCRP) with a supported monolayer containing a phosphorylcholine moiety was studied. Three types of phospholipids were synthesized, each containing a insertion spacer of eight, six, or three atoms between the phosphorylcholine group and hydrophobic tail. By varying the length of the insertion spacer, we can vary the extension of the phosphorylcholine group from the membrane surface. By varying the monolayer composition, we can control the lateral distance between the exposed phosphorylcholine groups. Using the surface plasmon resonance technique (SPR), we demonstrated that the calcium-dependent binding of rCRP to the model membrane is governed not only by the ability of the ligand to access the binding pocket fully (spacer length), but also by lateral hindrance within the two-dimensional plane of the membrane. The value of the apparent binding constant was estimated by theoretical analysis, which is obviously dependent on the composition of the lipid mixture, and a maximum of (9.9 +/- 1.5) x 10(6) M-1 was obtained.

A stereological study of the numbers of germ cells in various stages of spermatogenesis was undertaken in testosterone-treated intact and hypophysectomized (HPX) rats. Adult Sprague-Dawley rats were given testosterone by Silastic implants, which either inhibited (3-cm length) or partially maintained (10 cm) spermatogenesis over a 13-week period. The numbers of nuclei of the various germ cell categories (spermatogonia, spermatocytes, and round spermatids) in the testes were estimated by profile counting and measurement of nuclear diameter. The numbers of elongated spermatids were determined separately in testicular homogenates. Testis weight, seminiferous tubule volume, and tubule diameter were significantly decreased in intact rats with 3- and 10-cm testosterone implants and in HPX rats, although they were partially maintained in groups with 10-cm implants compared to those in groups with 3-cm implants (P less than 0.05). The effect of 3-cm testosterone implants in the intact group was to suppress the number of spermatogonia to 57%, reduce the conversion of spermatogonia to spermatocytes to 85%, and reduce the conversion of round to elongated spermatids to 19% of the control value. This latter effect was largely overcome with 10-cm testosterone implants. In HPX rats, only 10-cm implants were effective in maintaining the conversion of round spermatids to elongated spermatids. However, testosterone alone was less effective in maintaining the conversion of spermatocytes to round spermatids, suggesting that a pituitary factor, probably FSH, was involved. It is concluded that testosterone has a major effect on the conversion of round to elongated spermatids. The conversion of spermatogonia to spermatocytes and the conversion of spermatocytes to round spermatids depend on the synergistic action of both FSH and testosterone. However, the effect of FSH is greatest on the conversion of spermatocytes to spermatids, i.e. meiosis.

The Raman spectroscopic lines of liquid cultures of Rhizobium japonicum have been compared with electron microscopic examinations and growth measurements of these cells. The results showed that the significant Raman lines are related to the reproduction activities of the procaryotic cells.

Gossypol treatment of endometriosis and uterine myomas in 15 cases was started with a daily dose of 20 mg of gossypol-acetic acid twice a day for 20 days as a loading dose, followed by 40 mg (20 mg twice weekly). A maintenance dose of 20 mg was given weekly. The total course of treatment lasted about 6 months. Light and electron microscopic examination of the endometrium was carried out before treatment and 2, 3, 4, and 6 months after gossypol treatment. After gossypol treatment three different characteristics of the endometrium were noted: (1) irregular secretory activity, (2) proliferative activity, and (3) atrophy. On electron microscopic examination it was shown that there were important changes of the organelles, indicating suppressed secretory activity. Based on the above results, the nature and significance of the ultrastructural changes of the endometrium induced by gossypol were analyzed and the possibility of developing a new field of research in female contraception with gossypol was considered.