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 systematically evaluate the efficacy and safety of ciprofol for sedation and anesthesia outside the operating room.Methods Databases such as PubMed,Embase,Cochrane Library,Web of Science,CNKI,Wanfang Data,CBM,and VIP were searched for randomized controlled trials(RCTs)related to the efficacy and safety of ciprofol for sedation and anesthesia outside the operating room.The search covered all publications up to June 2023.Statistical analysis was performed using RevMan 5.4 software and Stata 15.0.Results Twelve RCTs were included,involving 2 192 patients,of which 1 154 were in the ciprofol group and 1 038 in the propofol group.Compared with the propofol group,the anesthesia induction time(MD=0.28 min,95％CI 0.08-0.47 min,P=0.006)and recovery time(MD=1.16 min,95％CI 0.44-1.87 min,P=0.001)were significantly longer in the ciprofol group,and the inci-dences of injection pain(OR=0.04,95％CI 0.02-0.06,P＜0.001),hypotension(OR=0.64,95％CI 0.49-0.83,P=0.0008),hypoxemia(OR=0.44,95％CI 0.21-0.91,P=0.03),and respirato-ry depression(OR=0.19,95％CI 0.11-0.32,P＜0.001)were significantly lower.There were no sta-tistically significant differences between the two groups in terms of sedation success rate,physician satisfac-tion,the difference in heart rate before and after anesthesia induction,incidence of body movement,brady-cardia,nausea and vomiting,and dizziness.Conclusion The anesthetic effect of cyclopofol and propofol is similar when used for anesthesia outside the operating room.Compared to propofol,ciprofol offers comparable anesthetic effects for sedation and anesthesia outside the operating room,with a lesser impact on respiratory function and more stable hemodynamics.Ciprofol also significantly lowers the incidence of adverse reactions such as injection pain,hypotension,hypoxemia,and respiratory depression.

Objective:To observe the effects of overexpression of S100A4 protein on retinal capillary cells and retinal ganglion cells (RGC) after retinal ischemia-reperfusion injury (RIRI).Methods:One hundred healthy adult male C57BL/6 mice were randomly divided into normal control group (group C), RIRI group, adeno-associated virus (AAV2)-S100A4 green fluorescent protein (GFP) intravitreal injection group (group S), RIRI+AAV2-GFP intravitreal injection group (group GIR), and RIRI+AAV2-S100A4-GFP intravitreal injection group (group SIR), with 20 mice in each group. The RIRI model was established using the high intraocular pressure anterior chamber method in the RIRI, GIR and SIR groups of mice. Eyes were enucleated 3 days after modelling by over anaesthesia. The number of retinal capillary endothelial cells and pericytes in the retinal capillaries of mice in each group was observed by retinal trypsinised sections and hematoxylin-eosin and periodic acid-Schiff staining; immunofluorescence staining was used to observe endothelial cell, pericyte coverage and RGC survival; The relative expression of Toll-like receptor 4 (TLR4), p38 MAPK and nuclear factor erythroid 2-related factor 2 (NRF2) in retinal tissues was measured by Western blot. One-way analysis of variance was used to compare data between groups.Results:Three days after modeling, the endothelial cell to pericyte ratio in group C was compared with group S and SIR, and the difference was not statistically significant ( F=106.30, P>0.05); the SIR group was compared with group RIRI and GIR, and the difference was statistically significant ( F=106.30, P<0.000 1). Comparison of endothelial cell coverage in each group, the difference was not statistically significant ( F=3.44, P>0.05); compared with the pericyte coverage in group C, the RIRI group and the GIR group were significantly lower, and the difference was statistically significant ( F=62.69, P <0.001). Compared with the RGC survival rate in group C, it was significantly lower in RIRI and GIR groups, and the difference was statistically significant ( F=171.60, P<0.000 1); compared with RIRI and GIR groups, the RGC survival rate in SIR group was significantly higher, and the difference was statistically significant ( F=171.60, P<0.000 1). The relative expression levels of TLR4, p38 and NRF2 proteins were statistically significant among all groups ( F=42.65, 20.78, 11.55; P<0.05). Conclusions:Pericytes are more sensitive to ischemia than endothelial cells after retinal RIRI in mice, and early vascular cell loss is dominated by pericytes rather than endothelial cells. The overexpression of S100A4 protein protects against loss of pericytes and RGC after RIRI by inhibiting the TLR4/p38/NRF2 signaling pathway.

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 analyze the influencing factors of mechanical phlebitis(MP)during peripherally inserted central catheterization(PICC)in very/extremely low birth weight infants(VLBWI/ELBWI)in neonatal Intensive Care Unit(NICU).Methods The clinical data of VLBWI/ELBWI who underwent PICC catheterization were retrospectively analyzed.Multivariate Logistic regression model was used to analyze the influencing factors of MP.Results The incidence of MP after implementa-tion of PICC in VLBWI/ELBWI was 26.1％(116/444).Univariate analysis showed that the position of catheter tip,catheter vein,gestational age,body mass at birth,gestational age when cathetering,body mass when cathetering,puncture times and length were influencing factors for MP in VLBWI/ELBWI(P＜0.05).Multivariate Logistic regression analysis showed that abnormal position ofcathe-ter tip(OR=2.526;95％CI,1.532 to 4.164)and puncture times ≥ 2 times(OR=1.952;95％CI,1.017 to 3.747)were independent risk factors.Conclusion The abnormal catheter tip position and the number of puncturing ≥2 times increase the risk of mechanical phlebitis in VLBWI/ELBWI after PICC.Early intervention should be taken as soon as possible to avoid the occurrence of MP so as to relieve infants'pain and improve the quality of clinical care.