Objective To explore the regulation mechanism of autophagy-related protein, microtubule-associated protein 1 light chain 3 (LC3), via c-Jun in methotrexate resistant human choriocarcinoma JEG-3 cell lines. Methods Human choriocarcinoma JEG-3 cell lines, and methotrexate resistant choriocarcinoma JEG-3 (JEG-3/MTXR) cell lines were used in our present study. Phosphorylation c-Jun (p-c-Jun) was evaluated after exposure to 0.02 ng/ml methotrexate for 72 hours in both cells by western blot. c-Jun gene was knockdown by small interference RNA (siRNA) in JEG-3/MTXR cells, and LC3 was evaluated by western blot and reverse transcription-PCR. The binding of LC3 promoter with c-Jun protein was detected via chromatin immunoprecipitation assay (ChIP) with or without 0.02 ng/ml methotrexate exposure. Results The results showed that p-c-Jun was up-regulated after methotrexate treatment for 72 hours (1.99±0.20, versus 0.20±0.06 at 0 hour;P<0.05) by western blot analysis in JEG-3/MTXR cell lines. Further investigation demonstrated that c-Jun-siRNA could inhibit the up-regulation of LC3 formation and after methotrexate exposure (LC3 mRNA:1.24±0.17 versus 3.03±0.43;LC3 protein:0.52±0.07 verus 1.20± 0.15; all P<0.05). The binding of LC3 promoter by c-Jun protein was up-regulated after methotrexate treatment by the method of ChIP in methotrexate resistant JEG-3/MTXR cells [(2.95 ± 0.35) times]. Conclusion Autophagy-related gene LC3 expression regulated by c-Jun protein may be involved in the effect mechanism of the development of methotrexate resistance in choriocarcinoma JEG-3 cells.

Objective To investigate the age distribution and types of human papillomavirus (HPV) cervical infection in patients in Wuhan region, so as to provide scientific basis for the prevention and treatment of HPV infection and cervical cancer.Methods 9 915 exfoliated cervical cell specimens from patients in Wuhan Medical and Health Center for Women and Children between January 2015 and March 2016 were performed HPV genotyping (21 subtypes), clinical data of 1 732 HPV positive cases were statistically analyzed.Results HPV-positive rates in ≤25, 26~, 36~, 46~, and ≥56 age groups were 22.15%, 15.90%, 17.04%, 19.97%, and 17.57%,respectively (χ2=36.587,P<0.01),HPV-positive rates in ≤25 and 46~ age groups were both higher than other age groups.There were significant differences in single infection and multiple infection rates among different age groups respectively(χ2=14.39, 36.51,respectively, both P<0.05),single infection rate was highest in 46~ and ≤25 age groups (15.41% and 15.24% respectively);multiple infection rates was highest in ≤25 and ≥56 age groups (6.90% and 5.86% respectively).The percentage of single infection and multiple infection were 75.58% and 24.42% respectively, the major single infection type was HPV high-risk subtype (84.34%),the major multiple infection types were high-risk and high-risk compound subtype as well as high-risk and low-risk compound subtype, accounting for 60.52% and 38.77% respectively.The main single infection types were HPV high-risk subtypes 33, 68, 31 and 16, as well as and low-risk subtype 11, the ratios of single infection to multiple infection were 3.13, 2.03, 1.71, 1.67 and 2.00 respectively.Conclusion Cervical infection rates in women in Wuhan region is high in ≤25 and 46~ age groups, there are differences in the distribution of different HPV subtypes of single infection and multiple infection.

Background Posterior capsular opacification(PCO) is common complication after extrecapsular extract of cataract.Matrix metalloproteinases-3 (MMP-3) can degrade all the extracellular matrix except polyose.The gene therapy of PCO upon MMP-3 is the researching hot topic.Fibronectin ( FN ) is a degrade gelatin,so its expression can reflect the effect of MMP-3 on LECs indirectly. Objective The aim of this study was to construct MMP-3 eukaryotic recombination plasmid and transfect to lens epithelium cells(LECs) for the observation of MMP3 expression,and to explore the feasibility of gene therapy for after cataract. Methods Six fresh lenses were obtained from pigs.LECs were cultured using explant method.The eukaryotic expression vector pEGFP-N1-MMP-3 was reconstructed with MMP-3 and pEGFP-N1 plasmids.The accuracy of MMP-3 gene fragment was confirmed by double enzyme digestion and DNA sequencing analysis.After transfecting pEGFP-N1-MMP-3 into LECs of pig,the expression of MMP-3 protein in the cells was indirectly observed by green fluorescent protein.The expression of FN in LECs was detected using Western blot. Results The result of double enzyme digestion was consistent with the base number of pEGFP-N1 plasmids and target fragment.By enlacing the result of DNA sequencing analysis with software,the resemblance of the DNA sequence of MMP-3 from recombination plasmid pEGFP-N1-MMP-3 and that of homo MMP-3 was 99.6％,indicating that the target fragment was inserted to pEGFP-N1 plasmids successfully.Green fluorescence for GFP was seen in the LECs in pEGFP-N1-MMP-3 transfected group,but absent response for GFP was in empty vector group.Western blot revealed that the relative expression level of FN in LECs was 0.666±0.008 in pEGFP-N1-MMP-3 trasfected group and 0.326 ±0.071 in empty vector group,with a significant difference between these two groups(P=0.000). Conclusions Eukaryotic recombination plasmid pEGFP-N1-MMP-3 is successfully constructed,and MMP-3 can be expressed in LECs after transfected.These results lay a foundation for the further research of MMP-3 gene therapy for PCO.

Objective To determine risk factors for the miss rate of colorectal adenomas during colonoscopy.Methods A total of 981 patients,diagnosed as having at least one polyp in colonoscopy,received a second colonoscopy in 6 months from November 2012 to March 2016.All polyps were removed in the second colonoscopy.Bio-information of patients such as sex,age,surveillance interval and features of polyps such as number,size,shape,location,pathology,withdrawal time,bowel preparation was retrospectively analyzed.Factors associated with the miss rates in these patients were analyzed with Chisquare and was also analyzed with Logistic regression model for multiple factors.Results A total of 981 patients were selected according to the inclusion and exclusion criteria,including 604 males and 377 females.Miss rates of males and females were 38.9％ (235/604) and 27.9％ (105/377) (P＜0.01)respectively.Age ranged from 25 to 87 years with mean age being 61.0±9.7 years.Miss rates of senior patients ＜65 and ≥65 years were 31.5％(195/619) and 40.1％(145/362) respectively (P＜0.01).A total of 1 728 adenomas were found in first colonoscopy.A total of 2 267 adenomas were found in the second colonoscopy.The adenoma miss rate was 23.8％ (539/2 267).The miss rate of adenoma whose size ≤ 5 mm was 42.5％ (311/732);and that of larger size of 6 to 9 mm was 17.8％ (194/1 090);that of even larger size,i.e.,≥10 mm,was 7.6％(34/445)(P＜0.01).Miss rates of Is,Isp,Ip,LST and Ⅱ adenomas in shape were 28.4％(489/1 720),9.3％(24/235),6.6％ (12/182),9.0％(6/67) 20.5％ (8/39) respectively (P＞0.05).Location with highest adenoma miss rate were descending colon,ascending colon and transverse colon,27.8％(64/230),25.5％(120/470),25.5％ (161/632) respectively.Miss rates of high and low risk adenoma were 44.8％ (277/618) and 17.4％ (63/363) (P＜0.01).The highest adenoma miss rate of all the pathology type was tubular adenoma.The adenoma miss rate was 26.9％ (449/1 671) (P＜0.01).Miss rates of good and poor bowel preparation were 30.2％ (271/897) and 82.1％ (69/84) (P＜0.01).Miss rates of adequate and inadequate withdrawal time were 24.3％ (174/717) and 62.9％ (166/264) (P＜0.01).Conclusion Male,old-age,diameter ≤ 5 mm,poor bowel preparation and inadequate withdrawal time,high risk adenoma are the risk factors for missed adenoma.But the shape and location of adenoma are not the risk factors.

This study tested the effects of the gastrointestinal pulse train electrical stimulation with different parameters and at different locations on the neuronal activities of the lateral hypothalamus area (LHA) in obese rats in order to find the optimal stimulation parameter and location. Eight gastric electrical stimulations (GES) with different parameters were performed and the neuronal activities of gastric-distension responsive (GD-R) neurons in LHA were observed. The effects of stimulations with 8 parameters were compared to find the optimal parameter. Then the optimal parameter was used to perform electrical stimulation at duodenum and ileum, and the effects of the duodenal and ileac stimulation on the GD-R neurons in LHA were compared with the gastric stimulation of optimal parameter. The results showed that GES with the lowest energy parameter (0.3 ms, 3 mA, 20 Hz, 2 s on, 3 s off) activated the least neurons. The effects of GES with other parameters whose pulse width was 0.3 ms were not significantly different from those of the lowest energy parameter. Most gastric stimulations whose pulse width was 3 ms activated more LHA neurons than the smallest energy parameter stimulation, and the effects of those 3 ms gastric stimulations were similar. Accordingly, the lowest energy parameter was recognized as the optimal parameter. The effects of stimulations with the optimal parameter at stomach, duodenum and ileum on the LHA neuronal activities were not different. Collectively, gastrointestinal electrical stimulation (GIES) with relatively large pulse width might have stronger effects to the neuronal activities of GD-R neurons in LHA of obese rats. The effects of the GIES at different locations (stomach, duodenum and ileum) on those neurons are similar, and GES is preferential because of its easy clinical performance and safety.

Objective To investigate the functional and histological changes in aging gerbil bladders.MethodsTwelve male gerbils were randomly divided into control group and aging bladder group,with 6 in each group.Experiment was conducted in gerbils of control group and aging bladder group 6 months and 24 months after normal feeding,respectively.Urodynamic examinations including irrigation volume,compliance and filling pressure of bladder were performed,bladder weight was obtained,bladder weight index was calculated,morphological observation was conducted with HE staining,ratio of amount of smooth muscle to collagen was obtained with double immunohistochemical staining,and electron microscope was used to evaluate the ultrastructure of bladder.ResultsCompared with those in control group,the irrigation volume and compliance of bladder significantly decreased in aging bladder group(P0.05).However,the bladder weight index was significantly lower in aging bladder group(P0.05).There existed degeneration in smooth muscle cells and organelles in aging bladder group.ConclusionThe function of aging bladder in gerbils is impaired,which may be related to the degeneration of bladder tissues.

AIM: To observe the effect of Ahmed glaucoma valve implantation intravitreal bevacizumab in the treatment of neovascular glaucoma ( NVG) .
METHODS:Twenty-two cases (22 eyes) who presented with NVG were first treated with intravitreal bevacizumab 0. 1mL ( 2. 5mg ), then with Ahmed glaucoma valve implantation after regression of iris neovessels. Cases were followed - up for 6 - 36 ( mean 24 ) mo with observation on visual acuity, IOP control, regression of iris neovessels, and complications during or after surgery.
RESULTS: Iris neovessels was regressed in different degree after injection within 1wk in 22 eyes. At final follow-up, the IOP of 18 eyes were all less than 21mmHg without any drugs and of 3 eyes with 1-3 kinds of anti-glaucoma drugs after combined Ahmed glaucoma valve implantation. The IOP of one eye was controlled after cryotherapy. The mean IOP dropped from 45. 36 ±8.13mmHg preoperatively to 15. 59 ± 3. 21mmHg postoperatively. IOP reduction was statistically significance between preoperative and postoperative ( P<0. 05) at final follow-up. Visual acuity was improved in 9 eyes (41%) and was no changed in 13 eyes. No serious complications were observed during or after intravitreous bevacizumab injection and Ahmed glaucoma valve implantation.
CONCLUSION: Ahmed glaucoma valve implantation and intravitreal bevacizumab in the treatment of NVG is useful and safe. It improves the success rate of surgery and preserves visual function, furthermore its complications are less.

With inlet temperature, specific gravity, feeding speed as independent variables, the comprehensive evaluating indexes of content of schisandrin and arctiin as dependent variable, the experimental data were fitted to a second order polynomial equation. Based on establishing the mathematical relationship between the comprehensive evaluating indexes and respective variables, Box-Benhnken central composite test and response surface analysis method was employed to optimize the spray drying technology of Biqiu granules ethanol extract. The optimal drying parameter was as follows: the inlet temperature was 175 degrees C, the specific gravity was 1.10, feeding speed was 32 r x min(-1). Under these conditions, the comprehensive evaluating indexes of spraying dry processes was 92.68, which was close to the model prediction. The spraying dry technology of Biqiu granules ethanol extract optimized by response surface methodology was accurate and feasible, which provided theoretical experiment basis for the industrialization production.
Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ethanol

Objective To investigate the relatonahip between TNF-α and ventricular arrhythmias after acute myocardial infarction(AMI)and its mechanism.Method Both the clinical and animal experiments were done.(1)Clinical experiment:Eighty patients with AMI were included in Union Hospital,Tongji Medical College,Huazhong University of Science and Techology,from May 2005 to November 2006 according to the WHO diagnostic criteria.Co-infection of diseases such as severe upper respiratory infection,lung infection,high fever,cancer,et al were excluded.The relationship between the levels of TNF-α and arrhythmias were observed at different times after AMI.A straight line correlation,analysis Was done.(2)Animal experiment:Different concentrations of TNF-αwere added to isohted rat hearts for observing the arrhythrnia effects.The effect of TNF-α on intracellular Ca2+ concentration was detected by laser confocal technique.All data were analyzed by SNK-q test using SPSS 13.0 sofeware prograrn.Results(1)The plasma levels of TNF-α were significantly associated with the Lown class of PVC after AMI and they were higher in AMI of anterior wall[(46.41±10.34)pg/ml]than other positions [(28.25±6.35)pg/ml,P<0.05].2)The frequency of ventricular arrhythmias was interrelated with the concentralions of TNF-α.Using etanercept beforehand,TNF-α induced a slight increase of intracellular Ca2+ intensity (P<0.05).Conclusions There was a relationship between TNF-αlevels and ventricular arrhythmisa in patients with AMI.Animal experiments confirmed the isolated heart perfusion with TNF-α induced ventricular arrhytrnias.Expression of TNF-α after AMI was related with the occurrence of ventricular arrhythrnias.The effect might be associated with the increased inuaeellular Ca2+ intensity caused by TNF-α.

First with the qualified rate of granules as the evaluation index, significant influencing factors were firstly screened by Plackett-Burman design. Then, with the qualified rate and moisture content as the evaluation indexes, significant factors that affect one-step pelletization technology were further optimized by Box-Behnken design; experimental data were imitated by multiple regression and second-order polynomial equation; and response surface method was used for predictive analysis of optimal technology. The best conditions were as follows: inlet air temperature of 85 degrees C, sample introduction speed of 33 r x min(-1), density of concrete 1. 10. One-step pelletization technology of Biqiu granules by Plackett-Burman design and Box-Behnken response surface methodology was stable and feasible with good predictability, which provided reliable basis for the industrialized production of Biqiu granules.
Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Temperature