Objective To investigate the inhibition of pathogenic viral gene expression in HPV6bE6-positive cell line by specific siRNA, which might have great potential for clinical use. Methods B16 cells were transfected with recombinant plasmid pcDNA3.1(+)-GFP/HPV6bE6, and the positive cell clones were selected by fluorescence protein observation and RT-PCR. Four specific siRNAs, none of which shares homology with exons of known human genes, were designed and synthesized to target HPV6bE6 mRNA. Quantitative real-time PCR was performed to measure the inhibition rates of target gene expression by comparing HPV6bE6 mRNA concentrations before siRNA transfection with those after transfection. The inhibition rates of target gene expression with different siRNA concentrations of 0.2 nmol/L, 1 nmol/L, 10 nmol/L, 50 nmol/L, 150 nmol/L and with different treatment time at 24 h, 48 h, 72 h and 96 h after transfection were measured respectively. Results More than 90% reduction of HPV6bE6 mRNA was observed following treatment with HPV6bE6-siRNA, and HPV-negative cells were apparently unaffected by HPV6bE6-siRNA. The decrease of HPV6bE6 mRNA was maximal at 24 h after siRNA treatment and sustained for at least 4 days. The minimal level of siRNA to efficiently silence the homogeneous target gene HPV6bE6 was 1 nmol/L. Conclusion HPV6bE6-siRNA can efficiently and specifically silence target genes and may be developed as a potential therapeutic approach for HPV infection.

A method was developed to assay α-Hydroxyltriazolam and α-Hydroxyalprazolam,　which are the major metabolites of triazolam and alprazolam respectively,in human urine.　After addition of 2-hydroxyflurazepam　(interal standard) and hydrolysis with β-glucuronidase, the hydroxy-metabolites were extracted with hexane-dichloromethane (1∶1) at pH　10.8, then were derivated with (BSTFA). The analysis was performed on a HP-5 capillary column with electron-capture detector.The detection limits of analysis in urine were about 1μg/L.The method was successfully applied to urine specimens collected from healthy human volunteers who ingested 0.5 mg of triazolam or 0.8 mg alprazolam. The method was enough sensitive to assay urine specimen excreted at 24 h by volunteers after taking the medicine.

Objective:To investigate the effect of heat shock protein B8 (HspB8) downregulation on retinal ganglion cell (RGC) and retinal function in the mice model of optic nerve injury (ONC).Methods:Adeno-Associated Virus (AAV) 2 AAV2-shHspB8-GFP was constructed to knockdown HspB8. 66 adult male C57/BL6 mice were randomly divided into the control group, the ONC group, the AAV2-shHspB8 group, the ONC+AAV2-shHspB8 group, and the ONC+AAV2-GFP group. There were 10, 20, 16, 10 and 10 mice respectively, and both eyes were used as experimental eyes. Western blot was used to evaluate the expression of HspB8 on day 3 and 7 after ONC. By GFP immunofluorescence staining, the efficacy of AAV2-shHspB8-GFP transfer was accessed. Moreover, it was possible to identify functional and RGC survival differences between groups by optomotor response (OMR), dark adapted full-field flash electroretinogram (ff-ERG), oscillatory potentials (OPs), photopic negative response (PhNR) and retinal flat-mount RGC counting 5 days after ONC. Comparisons between two groups were made using Mann-Whitney U test, unpaired t-test, unpaired t-test with Welch’s correction, one-way ANOVA, and Bonferroni t test. Results:Compared with the control group, the expression of HSPB8 protein in the retina of mice in ONC3 group was significantly increased, and the difference was statistically significant ( F=43.63, P<0.01). Compared with the control group, the ONC group showed obviously lower visual acuity ( P<0.01), lower a-wave, b-wave, OPs, PhNR amplitude, longer b-wave latency ( P<0.05), and the survival rates of RGC in ONC3 group, ONC5 group and ONC7 group decreased in a time-dependent manner( F=384.90, P<0.01). Transfection of AAV2 efficiency was highest on 4 weeks after IVT. Besides, there was no significant differences between the control group and the AAV2-shHspB8 group on visual acuity, ff-ERG, OPs, PhNR and RGC survival ( P>0.05). In comparison of the control group, we found that RGC survival of the ONC5+AAV2-shHspB8 group was significantly elevated ( F=10.62, P<0.01). Conclusions:Expression of HspB8 on the retina can be induced by ONC. The investigation of RGC counting, visual acuity, and ff-ERG revealed that optic nerve injury destructed functionality of mice retina and resulted to RGC death ultimately. The Most crucial finding of this research is that HspB8 knockdown had a neuroprotective effect in RGC after ONC.

Objective:To investigate the effects of Krüppel-like factor 7 (KLF7) on the survival of retinal ganglion cells (RGCs) and electroretinogram (ERG) after retinal ischemia-reperfusion (RIR) injury in mice.Methods:A total of 126 male C57BL/6J mice were randomly divided into normal group, RIR group, normal-KLF7 group, normal-green fluorescent protein (GFP) group, RIR-KLF7 group and RIR-GFP group. At the age of 8 weeks, mice of normal-KLF7 group and RIR-KLF7 group were intravitreally injected 1ul of 1.0×10 12 vg/ml adeno-associated virus overexpressing KLF7 (AAV2-KLF7-GFP). Mice of normal-GFP group and RIR-GFP group were injected adeno-associated virus of AAV2-GFP with the same titer. At the age of 11 weeks, RIR injury was induced in mice of RIR group, RIR-KLF7 group and RIR-GFP group, and intraocular pressure was measured. Retinal cryosections were used to access the efficacy of virus transfection 4 weeks after AAV2-KLF7-GFP transfer. 7 days after RIR injury, RGCs' survival rate was observed and quantified by immunofluorescent staining. ERG was performed to observe the differences in amplitudes and incubation period of scotopic ERG a-, b-wave, oscillatory potentials (Ops), photopic negative responses (PhNR). Optomotor response was performed to observe the differences of visual acuity. Expression of KLF7 was detected by western blot 4 weeks after AAV2-KLF7-GFP transfer. Results:Compared with normal group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR group were decreased, and the differences were statistically significant ( t=12.860, 7.157, 5.735, 8.953, 4.744, 9.887; P<0.05). With the increase of light intensity, the amplitudes of scotopic ERG a- and b-wave were gradually increased while the incubation period was gradually shortened. Compared with RIR group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR-KLF7 group were increased, and the differences were statistically significant ( t=6.350, 3.253, 3.695, 5.825, 5.325, 4.591; P<0.05). Protein level of KLF7 was up-regulated in normal-KLF7 group than those in normal group, and the difference was statistically significant ( t=4.105, P<0.01). Conclusion:Overexpression of KLF7 can improve RGCs’ survival rates and preserve the electrophysiological function.

In recent years, artificial intelligence (AI) and oncology are rapidly integrated and developed, gradually moving towards clinical application. In China, the incidence and mortality of gastric cancer rank first among various tumors. The development of AI technologies such as convolutional neural network, narrow-band imaging magnifying endoscope, full-field digital section and survival circulation network has the potential to be applied to early screening, auxiliary diagnosis, pathological diagnosis, prognosis analysis, as well as individualized diagnosis and treatment process of gastric cancer. However, AI medical treatment still has many problems such as insufficient data sample size and difficulty in integration. Reasonable design and application of AI technology is expected to provide more scientific and accurate diagnosis and treatment for patients with gastric cancer, improve the overall survival rate and quality of life of patients with gastric cancer, and achieve more breakthroughs in the medical field.

OBJECTIVE:To determine quality indexes of Hejiang Dendrobium nobile at different harvesting time and months,and to provide scientific reference for reasonable harvesting period of Hejiang D.nobile.METHODS:The stem of annual,biennial and triennial D.nobile were collected.The drying rate,the contents of polysaccharides (colorimetry),dendrobine (GC) and total alkaloids (colorimetry),effective component total rate (dendrobine×drying rate) were measured.Biennial D.nobile were collected in autumn and winter (Oct.of the second year-Mar.of the third year,15th day a month) to determine drying rate,the contents of polysaccharides and dendrobine.RESULTS:The drying rate,the contents of polysaccharides and effective component total rate were higher than annual and triennial D.nobile.The content of total alkaloids and dendrobine were in descending order:annual D.nobile (0.52％,0.48％) ＞biennial D.nobile(0.48％,0.44％) ＞triennial D.nobile (0.32％,0.22％).From Oct.of the second year to Mar.of the third year,the drying rate of biennial D.nobile was increasing month by month;the contents of polysaccharides and dendrobine increased firstly and then decreased;the content of polysaccharide was the highest in Feb.(17.32％),and the content of dendrobine reached the highest level in Dec.(0.51％).CONCLUSIONS:The optimal harvesting period is biennial Hejiang D.nobile in Dec.and Jan.(third years before flowering),considering flowering characteristics,drying rate,the contents of polysaccharide and dendrobine.

This review describes the cognitive function impairment and recovery during the active and remission phases of Cushing’s syndrome (CS) , a glucocorticoid over-exposure disease. Memory, attention, executive ability, and intelligence disorders are included. A comprehensive view of the present studies on the CS-related cognitive functions provides new ideas for a simpler and full-scale psycho-cognitive assessment of neuroendocrine diseases.

OBJECTIVE： To study the chemical constituents from the petroleum ether part from whole herbs of Pteris vittata. METHODS： The petroleum ether part from whole herbs of P. vittata was separated and isolated by silica gel column， gel column， recrystallization and TLC. The structures of the compounds were identified according to physicochemical properties and spectrum data （1H-NMR and 13C-NMR）. RESULTS： A total of 11 compounds were isolated and identified from the petroleum ether part from whole herbs of P. vittata， as （2R）-acetyl pterosin B （Ⅰ）， palmitic acid （Ⅱ）， hop-22（29）-ene （Ⅲ）， epifriedelanol （Ⅳ）， lupenone （Ⅴ）， olean-18-en-3-one （Ⅵ）， stigmasterol （Ⅶ）， β-sitosterol （Ⅷ）， 22-hydroxyhopane （Ⅸ）， ergosterol （Ⅹ）， β-sitosterol acetate （Ⅺ）. CONCLUSIONS： Compounds Ⅰ-Ⅶ and Ⅸ-Ⅺ are isolated from this plant for the first time， and can provide theoretic reference for further studying bioactive pharmacodynamic substances in P. vittata and enriching chemical component data.

ObjectiveTo explore the drug resistance of Morganella morganii （Mm） and the risk factors for extended-spectrum β-lactamase （ESBL） positive infection in type 2 diabetes foot （DF） patients with Mm， and to provide a reference for clinical treatment. Methods310 samples of DF patients with Mm infection were collected from Shanghai Integrated Traditional Chinese and Western Medicine Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from January 2020 to March 2023. The patients were selected for the first time to detect Mm bacteria through cultivation， and were divided into ESBL producing group （n=45） and non ESBL producing group （n=265）. Bacterial identification and drug sensitivity experiments on Mm were conducted. Multivariate logistic regression analysis was used to analyze the risk factors. The stepwise regression method （SRM） was used to screen the most important correlation factors for constructing risk prediction model and its evaluation. ResultsBoth ESBL producing and non ESBL producing Mm were sensitive to imipenem （IPM） and meropenem （MEM）. Compared with the non ESBL producing group， the resistance of Mm in the ESBL producing group to other tested antibiotics ［excluding ampicillin （AM）/sulbactam （SU）］ was higher （P<0.05）. Aged ≥60 years old， concomitant hypoproteinemia （HP）， combined use of antibiotics， and history of third-generation cephalosporin use were all independent risk factors for the occurrence of ESBL producing strain infection （P<0.05）. SRM screening identified age， HP， and use of third-generation cephalosporins as the most associated factors with ESBL producing strain infection， and these three factors were included in the multivariate logistic regression prediction model. After calculation， when P=0.80， the Jordan index was the highest and the prediction effect was the best. The prediction accuracy was 89.35%， the sensitivity was 86.67%， and the specificity was 89.81%. The model evaluation results showed that the predictive model has good discrimination and high accuracy. ConclusionThe Mm strain producing ESBL has strong resistance to commonly used antibiotics， and it is recommended that clinical physicians use antibiotics reasonably. Age， HP， combined use of antibiotics， and use of third-generation cephalosporins are all independent risk factors for the occurrence of ESBL producing bacterial infections. Clinical monitoring should be carried out on susceptible populations based on risk factors， and infection management should be strengthened.

ObjectiveTo explore the mental health status and its influencing factors among middle school students in Xide County， Liangshan Yi Autonomous Prefecture， and to provide references for mental health interventions for local middle school students. MethodsUsing a cross-sectional study design， one junior middle school and one senior middle school in Xide County， Liangshan Prefecture， Sichuan Province， were randomly selected on September 16， 2021， and two classes per grade in each school involving 288 students were recruited. Subjects were assessed using Patients' Health Questionnaire Depression Scale-9 item （PHQ-9）， Generalized Anxiety Disorder Scale-7 item （GAD-7）， PTSD Checklist for DSM-5 （PCL-5）， Multidimensional Scale of Perceived Social Support （MSPSS） and UCLA Loneliness Scale （ULS-3）. Then the scores of above scales were compared among middle school students with different demographic characteristics， and binary Logistic regression analysis was conducted to screen the influencing factors of post-traumatic stress disorder (PTSD） symptoms. ResultsAmong the respondents， 17.71% （95% CI： 0.133~0.221）， 8.68% （95% CI： 0.054~0.120）， 2.78% （95% CI： 0.009~0.047） and 45.83% （95% CI： 0.400~0.516） were reported to have symptoms of depression， anxiety， PTSD and loneliness， respectively. Students in senior middle school scored lower on PCL-5 and ULS-3 than those in junior middle school ［（6.46±8.91） vs. （8.87±9.42）， t=2.202， P<0.05； （4.67±1.65） vs. （5.60±1.88）， t=4.431， P<0.01］. Regression analysis denoted that depressive symptoms （OR=7.630， P<0.05） and anxiety symptoms （OR=3.789， P<0.01） were risk factors for PTSD symptoms among middle school students. ConclusionThe middle school students in Xide County， Liangshan Yi Autonomous Prefecture suffer a high prevalence rate of depressive symptoms and loneliness， and those in junior middle school are more likely to feel a sense of strong loneliness， furthermore， depressive symptoms and anxiety symptoms are risk factors for PTSD symptoms.