Objective To preliminarily explore the potential efficacy of Xuesaitong Soft Capsule(XST)against post-stroke depression(PSD),and to investigate the material basis of XST's anti-PSD effect based on the metabolomics results to analyze its related pharmacokinetic(PK)characteristics and further analyze the pharmacodynamic(PD)equation of representative ingredients.Methods The initial evaluation of drug effica-cy was conducted by detecting the depressive-like behavior and neurotransmitter levels in rats.The Pearson correlation analysis was employed to analyze the correlation between the main metabolites regulated by XST and the saponin components entering the bloodstream.At various time points after drug administration,the blood concentration of ginsenoside Re and the concentration of norepinephrine(NE)in the serum of PSD rats were measured,and the compartment model was fitted accordingly.Furthermore,the liquid chromatography-mass spectrometry was utilized to determine the content of ginsenoside Re in the liver,spleen,kidney,prefron-tal cortex,hippocampus and striatum of PSD rats.Results Ginsenoside Re showed the optimal correlation by the Pearson correlation analysis.Based on its pharmacokinetic parameters,the pharmacodynamic equation with NE was E=160.462 × Ce/(38.663+Ce).The contents of ginsenoside Re in the liver,spleen,kidney,prefron-tal cortex,hippocampus and striatum of rats were(17.23+11.90),(19.05+5.67),(1.95+0.79),(70.13+6.75),(57.03+3.11),and(72.45+5.45)ng/g,respectively.Conclusion XST could improve the depressive-like behaviors in PSD rats by regulating the expression levels of neurotransmitter NE and 5-HT.Ginsenoside Re may be the pharmacodynamical material foundation for XST's preventative treatment of PSD.

Objective:To study the surveillance results of soil-transmitted nematode (STN) disease at the national surveillance site in Nanchang City, so as to scientifically formulate prevention and control strategies for parasitic diseases in Nanchang City.Methods:From 2016 to 2020, in Nanchang County, the national surveillance site of STN disease in Nanchang City, no less than 1 000 fecal samples and 25 soil samples of permanent residents over 3 years of age were investigated annually. The improved Kato-Katz thick smear method was used to detect the eggs of STN in the population, species identification of hookworm was carried out by test-tube filter paper incubation, the Enterobius vermicularis eggs of 3 - 9 years old children were detected by cellophane anal swab method, and the soil samples were used to identify hookworm larvae and of Ascaris lumbricoides eggs. Results:The total infection rate of STN was 0.93% (47/5 054) from 2016 to 2020 in the national surveillance site of Nanchang City. The annual infection rates were 0.10% (1/1 000), 2.94% (30/1 021), 0.79% (8/1 008), 0.50% (5/1 002) and 0.29% (3/1 023), respectively; the infection rates of hookworm, Ascaris lumbricoides and Trichuris trichiura were 0.42% (21/5 054), 0.02% (1/5 054) and 0.49% (25/5 054), respectively. Totally 97.87% (46/47) of the infected persons were mild infection. The population distribution characteristics showed that there was significant difference in STN infection rate among different education levels (χ 2 = 18.06, P = 0.001), but there was no significant difference in infection rate among different sex, age and occupation (χ 2 = 0.92, 2.01, 13.60, P > 0.05). Among them, the infection rate of junior high school cultural was the highest (1.84%, 24/1 301). The culture of hookworm larvae showed that 90.48% (19/21) were Necator americanus; the infection rate of Enterobius vermicularis in 3 - 9 years old children, the contamination rate of soil hookworm larvae and Ascaris lumbricoides eggs, were all 0. Conclusion:STN disease shows a low prevalence trend at the national monitoring site in Nanchang City, Trichuris trichiura and hookworm are the key insect species for STN disease control in Nanchang City in the future.

Background: Insulin regulates glucose homeostasis and has important effects on metabolism, cell growth, and differentiation. Depending on the cell type and physiological context, insulin signal has specific pathways and biological outcomes in different tissues and cells. For studying the signal pathway of insulin on glycolipid metabolism in porcine embryonic fibroblast (PEF), we used high-throughput sequencing to monitor gene expression patterns regulated by insulin. Objectives: The goal of our research was to see how insulin affected glucose and lipid metabolism in PEFs. Methods: We cultured the PEFs with the addition of insulin and sampled them at 0, 48, and 72 h for RNA-Seq analysis in triplicate for each time point. Results: At 48 and 72 h, 801 and 1,176 genes were differentially expressed, respectively. Of these, 272 up-regulated genes and 264 down-regulated genes were common to both time points. Gene Ontology analysis was used to annotate the functions of the differentially expressed genes (DEGs), the biological processes related to lipid metabolism and cell cycle were dominant. And the DEGs were significantly enriched in interleukin-17 signaling pathway, phosphatidylinositol-3-kinase-protein kinase B signaling pathway, pyruvate metabolism, and others pathways related to lipid metabolism by Kyoto Encyclopedia of Genes and Genomes enrichment analysis. Conclusions These results elucidate the transcriptomic response to insulin in PEF. The genes and pathways involved in the transcriptome mechanisms provide useful information for further research into the complicated molecular processes of insulin in PEF.

Objective:To explore the effect of pulmonary rehabilitation program based on Delphi method in patients with overlap syndrome (OS) .Methods:From March 2019 to March 2021, a total of 107 OS patients admitted to Qinghai Provincial People's Hospital were enrolled using the convenience sampling method and divided into the observation group ( n=54) and the control group ( n=53) by the random number table method. The control group received conventional nursing, and the observation group conducted pulmonary rehabilitation program based on Delphi method on the basis of conventional nursing. The COPD Assessment Test (CAT) , the 6-Minute Walk Test (6MWT) , and the St George's Respiratory Questionnaire (SGRQ) were used to assess the improvement of the conditions, exercise endurance, and quality of life of the two groups before and after intervention. Results:After intervention, the CAT score of the observation group was lower than that of the control group, 6MWD was longer than that of the control group, and the clinical symptoms, disease impact, and activity scores of SGRQ were lower than those of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The pulmonary rehabilitation program based on the Delphi method can reduce patients' condition, improve exercise endurance and quality of life.

Objective Summarize and share the working practice of Investigator Research Ethics and Research Integrity Training conducted at Peking University Human Research Protection Program (PKU HRPP) ,to further explore continuing quality improvement of investigator ethical training at university level .Methods Conduct systematic review of the archiving files of PKU HRPP investigator training activities during 2012-2018 ,summarize feedback information from investigators to i-dentify possible experiences for sharing and space for improvement .Results There are some positive experiences for sharing a-bout the training mechanism and practices at PKU HRPP .Conclusions Based on the previous ethical training work and experi-ences at PKU HRPP ,possible proposals for continuing quality improvement may including :strengthening the requirements of ethical training of investigators ,encouraging and recognizing ethical training conducted by research teams ,emphasizing training Quality and the promotion of sharing and mutual recognition mechanisms for ethical training .

The clustered regularly interspaced short palindrome repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a versatile genome editing tool with high efficiency. A guide sequence of 20 nucleotides (nt) is commonly used in application of CRISPR/Cas9; however, the relationship between the length of the guide sequence and the efficiency of CRISPR/Cas9 in porcine cells is still not clear. To illustrate this issue, guide RNAs of different lengths targeting the EGFP gene were designed. Specifically, guide RNAs of 17 nt or longer were sufficient to direct the Cas9 protein to cleave target DNA sequences, while 15 nt or shorter guide RNAs had loss-of-function. Full-length guide RNAs complemented with mismatches also showed loss-of-function. When the shortened guide RNA and target DNA heteroduplex (gRNA:DNA heteroduplex) was blocked by mismatch, the CRISPR/Cas9 would be interfered with. These results suggested the length of the gRNA:DNA heteroduplex was a key factor for maintaining high efficiency of the CRISPR/Cas9 system rather than weak bonding between shortened guide RNA and Cas9 in porcine cells.
Base Sequence ; Complement System Proteins ; CRISPR-Cas Systems ; DNA ; Genome ; Nucleotides ; RNA, Guide ; Swine

Objective To discuss key points of surgical cooperation in laparoscopic radical resection of high rectal cancer with transvaginal specimen extraction and without abdominal incision, so as to provide valuable experience and reference to better cooperation in this surgical method and other similar surgical methods. Methods Clinical data and surgical cooperation of 16 patients undergoing this surgical method were analyzed retrospectively, with key points of the operation nursing summarized. Results All the 16 patients tolerated general anesthesia and carbon dioxide pneumoperitoneum, which made smooth surgeries without conversion to laparotomy and adverse reactions. All the patients were cured and left the hospital with fine condition of recovery. Conclusions Full preoperative preparation of nurses, familiarity of surgical process, good command of laparoscope instruments, usage of special instruments, and rigorous non-bacteria and non-tumor technique operation, are all keys to a successful operation.

Objective:To investigate the effects of interleukin-17 on tumor,we transfected interleukin-17 gene into mouse colon cancer cells(C26)and set up an animal model in tumor.Methods:By plasmid vector,IL-17 gene was transfected into C26.Meanwhile, empty plasmid vector(pcDNA3.1)and C26 cells were transfected as control groups.C26/pcDNA3.1-IL-17,C26/pcDNA3.1,and C26 cells were subcutaneously inoculated into mice respectively and the tumor volume and the survival time were observed.Proliferation of splenocyte and NK activity were detected.Detect the characteristic cytokines and transcriptional factors of Th1,Th2,Th17 and Treg cells in splenic lymphocyte.Proliferation of TIL was detected.The characteristic cytokines IL-10 of M1 and the characteristic cytokines IL-12 of M2 in tumor infiltrating macrophages were detected.Results: The growth of tumor in mice inoculated with C26/pcDNA3.1-IL-17 cells was significantly retarded ( P<0.05 ) , and the growth of tumor in male mice inoculated with C26/pcDNA3.1-IL-17 cells was significantly retarded than female mice ( P<0.05 ).The mice survival time of C26/pcDNA3.1-IL-17 group was similar with C26/pcDNA3.1 and C26 groups(P>0.05).The proliferation of the splenocytes from C26/pcDNA3.1-IL-17 inoculated mice was higher than those of C26/pcDNA3.1,C26 groups(P<0.05),but was similar with the normal group(P>0.05),the proliferation of the splenocytes from C26/pcDNA3.1 and C26 inoculated mice was slow than those of normal groups(P<0.05).The NK(separate from spleen) activity of C26/pcDNA3.1-IL-17,C26/pcDNA3.1 and C26 inoculated mice was lower than those of normal groups when the ratios of effector cells and target cells were 40∶1,20∶1(P<0.05),the NK(separate from spleen) activity of C26/pcDNA3.1-IL-17 inoculated mice was higher than those of C26/pcDNA3.1 and C26 groups when the ratios of effector cells and target cells were 40∶1(P<0.05),there′s no difference among every groups when the ratio of effector cells and target cells were 10∶1 ( P>0.05 ).The splenocytes from the mice inoculated with C26/pcDNA3.1-IL-17 cells secreted more IFN-γ( the characteristic cytokines of Th1 ) , IL-4 ( the characteristic cytokines of Th2),GATA-3,ROR-γt,IL-10(the characteristic cytokines of Treg)mRNA(P<0.05).The proliferation of TIL from C26/pcDNA3.1-IL-17 inoculated mice was higher than those of C26/pcDNA3.1,C26 groups(P<0.05),the proliferation of TIL from C26/pcDNA3.1-IL-17,C26/pcDNA3.1 and C26 inoculated mice was lower than those of normal groups( P<0.05).And there′s no differences among every groups of the express of cytokines IL-10 and IL-12 mRNA in tumor infiltrating macrophages(P<0.05).Conclusion: The transfection of IL-17 gene inhibited tumor growth in the mice,inoculated with enhancing the immune function.

Von Hippel-Lindau (VHL) disease is an autosomal dominant disorder and its clinical manifestation including haemangioblastomas of the central nervous system, renal cell carcinoma, haeochromocytomas, and pancreatic cyst. The deletion, mutation and promoter methylation of VHL gene can cause VHL disease. Swine is considered as an ideal model for human disease because of its physiological and anatomical similarity to human. We cloned pig VHL gene that is 2 725 bp in length. VHL highly expressed in adrenal gland, liver, pancreas, heart and testis. We designed 5 shRNAs and screened the most effective interference RNA fragment with a knockdown efficiency of 72%. Porcine embryonic fibroblasts stably transfected with pGenesil-shRNA vector were used as donor cells for nuclear transfer and there was no significant difference of embryo development compared with the control group. Moreover, VHL was efficiently knocked-down with efficiency of 71% in porcine cloned blastocyst, these results lay a solid foundation for constructing the VHL knock-down model of pig.
Animals ; Cloning, Molecular ; Disease Models, Animal ; Embryo, Mammalian ; Gene Knockdown Techniques ; Swine ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics ; von Hippel-Lindau Disease ; genetics

Reversed phase high performance liquid chromatography with diode array detector was employed for simultaneous determination of five main index components and specific chromatograms analysis in Xiaochaihu granules with a linear gradient elution of acetonitrile-water (containing 0.1% phosphoric acid) as mobile phase. The results showed that five main index components (baicalin, baicalein, wogonoside, wogonin, enoxolone) were separated well under the analytical condition. The linear ranges of five components were 0.518 - 16.576, 0.069 - 2.197, 0.167 - 5.333, 0.009 - 0.297 and 0.006 - 0.270 mg x g(-1), respectively. The correlation coefficients were 0.999 9, and the average recoveries ranged from 95% to 105%. Twelve common peaks were selected as the specific chromatograms of Xiaochaihu granules with baicalin as the reference peak. There were good similarities between the reference and the ten batches of samples. The similarity coefficients were no less than 0.9. The analytical method established is highly sensitive with strong specificity and it can be used efficiently in the quality control of Xiaochaihu granules.