Aim To investigate the protective effects of huang lian jie du tang active fraction(HLJDTAF) on ischemia injury in cerebral cortical neuronal cultures. Methods The cortical neurons of rat fetal were cultured in vitro. The protective effects of HLJDTAF on ischemia injury were observed by treating cells with sodium dithionite in glucose free medium. Results HLJDTAF 610 mg?kg -1 increased the activity of neuron, 305 mg?kg -1 reduced the leakage rate of LDH; HLJDTAF 610, 305, 153 mg?kg -1 reduced the content of NO,MDA and increased the activity of SOD. Conclusion HLJDTAF protected the cerebral cortical neurons from ischemia injury by reducing the contents of NO and suppressing the generation of lipidperoxide.

Background and purpose: High expression of telomerase and telomere stability are two common features in tumor cell. hTERT is a catalytic subunit of telomerase, TRF2 is extremely important to maintain the length and stability of telomerase. This study was to construct the recombinant adenovirus mediated shRNA to hTERT and TRF2, and to investigate the inhibitory effects of the vector by solo-inhibiting and connect-inhibiting in the MCF-7 cells, in order to present a new approach to the gene therapy for breast cancer. Methods: rAd-hTERT and rAd-TRF2 were constructed and the expression of hTERT mRNA and TRF2 mRNA were tested by FQ-PCR 48 hours after transfecting in MCF-7 cells. Apoptosis was observed by flow cytometry 1 to 6 days after transfection. Results: ①At 48 hours after transfection, the results of FQ-PCR showed that compared to PBS group, the expression of hTERT in rAd-hTERT group was obviously decreased and the inhibition ratio was about 86%, but TRF2 had not been obviously inhibited (P>0.05);the expression of TRF2 in rAd-TRF2 group was obviously decreased and the inhibition ratio was about 80%, but hTERT had not been obviously inhibited (P>0.05);in rAd-hTERT/rAd-TRF2 group, the inhibition ratio of hTERT and TRF2 were about 88% and 85%. Comparing rAd-hTERT/rAd-TRF2 group with rAd-hTERT group and rAd-TRF2 group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). Otherwise, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). ②The result of flow cytometry showed that apoptosis was induced at the first day after transfecting in rAd-hTERT group and rAd-TRF2 group, the most obvious apoptosis was in the 3rd to 5th days,at the peak in the 5th day, and decreased in the 6th day after transfection. The apoptosis ratio of rAd-hTERT group was 46.2%, rAd-TRF2 group was 43.5%. The apoptosis ratio of rAd-hTERT/rAd-TRF2 group was 46.2% at first day, 68.5% at the second day, the most obvious apoptosis was in the 3rd to 6th days and was 77.6% in the 6th days in rAd-hTERT/rAd-TRF2 group. There were significant differences in apoptosis ratio in solo-inhibiting and connect-inhibiting(P<0.05). In addition, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences in apoptosis ratio(P>0.05). Conclusion: ①Target sequence of RNAi which aimed at hTERT gene and TRF2 gene was designed efficiently, and the RNAi expression vectors were seen in vivo study efficiently and specifically inhibited the correspond gene expression and promoted cell apoptosis in MCF-7 cells. ②rAd-hTERT vector and rAd-TRF2 vector have no synergistical effect and antagoinstical effect on inhibiting hTERT gene and TRF2 gene mRNA expressing in MCF-7, but there was synergistical effect in terms of the induction of apoptosis. So association-RNAi-technique targeting to the genes of telomere length and stability can effectively promote tumor cell into apoptosis and inhibit breast cancer cell growth. RNAi technique of connecting correlation genes is a more effective gene therapy strategy.

Objective:To assess if metagenomic next-generation sequencing (mNGS) of periprosthetic joint tissue can provide an alternative rapid and sensitive tool for the diagnosis of prosthetic joint infection (PJI), especially compared to microbiological culture.Methods:A total of 33 eligible patients who underwent revision arthroplasty from June 2019 to June 2020 in orthopedic surgery department of the first affiliated hospital of Zhengzhou University were retrospectively analyzed. Twenty-one patients were included in PJI group according to the American Academy of Musculoskeletal Infection diagnostic criteria, with 17 cases of knee and 4 cases of hip, including 9 cases of male and 12 cases of female, with an average age of 59.14±14.55 years old (range from 28 to 84), and an average BMI of 23.7±2.8 kg/m 2 (range from 17.7 to 29.4 kg/m 2). Twelve patients were included in aseptic loosening group (control group), with 4 cases of knee and 8 cases of hip, including 4 cases of male and 8 cases of female, with an average age of 53.08±10.05 years old (range from 39 to 70), and an average BMI of 25.2±2.9 kg/m 2 (range from 18.3 to 31.2 kg/m 2 ). Microbiological culture results of synovial fluid and periprosthetic joint tissue and mNGS results of periprosthetic joint tissue were collected. The sensitivity and specificity of mNGS and microbiological culture were calculated and compared. The species of pathogenic microorganismsdetected by the two techniques were summarized. In addition, the impact of antibiotic use on the efficacy of both techniques were compared. Results:mNGS detected 13 positive cases and microbiological culture detected 6 positive cases in the PJI group. In the aseptic loosening group, 1 case was determined positive by mNGS, and all the microbiological culture results were negative. In the diagnosis of PJI, mNGS showed significantly higher sensitivity than that of culture (61.9% vs 28.6%, χ2=4.71, P=0.03), while no statistical difference was observed in terms of specificity (91.7% vs 100%, χ2=1.04, P=0.31). In the PJI cases with prior exposure to antibiotics within two weeks, the sensitivity of mNGS was significantly higher than that of culture (53.8% vs 15.4%, χ2=4.25, P=0.04). However, there was no significant difference in the sensitivity between mNGS and culture in patients without antibioticsexposure (66.7% vs 44.4%, χ2=0.90, P=0.34). In the detection of pathogenic microorganism, mNGS detected 9 kinds of bacteria (Staphylococcus aureus, Staphylococcus family, Moraxella Oslo, Propionibacterium acnes, Streptococcus acnes, Staphylococcus epidermidis, Mycobacterium tuberculosis, Staphylococcus Lyons, Bacteroides fragilis) and 2 kinds of fungi (Aspergillus fumigatus, Candida parapsilosis), while microbiological culture detected 3 kinds of bacteria (Staphylococcus aureus, Moraxella catarrhalis, Mycobacterium tuberculosis) and one kind of fungi (Candida parapsilosis). mNGS and microbiological culture were both positive in 5 cases, among which 3 cases had completely matched results (Staphylococcus aureus, Mycobacterium tuberculosis, Candida parapsilosis), one case had partly matched results (mNGS detected more bacteria than culture) and one case had totally mismatched results. Additionally, in the diagnosis of the 3 included tuberculous PJI, mNGS showed 100% specificity and sensitivity. Conclusion:mNGS of periprosthetic joint tissue is a more powerful tool for diagnosis and pathogen detection of PJI compared to microbiological culture, especially in the diagnosis of tuberculosis PJI. Besides, mNGS is more resistant to antibiotic exposure than culture.

Adjuvants, Immunologic ; therapeutic use ; Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Drug Therapy, Combination ; Female ; Hepatitis B, Chronic ; drug therapy ; Humans ; Lamivudine ; therapeutic use ; Male ; Middle Aged ; Thymosin ; analogs & derivatives ; therapeutic use

Objective:To deeply explore the potential mechanism of Kangmin Zhisou Granules in the treatment of bronchial asthma through network pharmacology method; To verify it with animal experiments.Methods:The active components and corresponding target information of Kangmin Zhisou Granules were screened with the help of BATMAN-TCM database, and the related disease targets of bronchial asthma were obtained through GeneCards and OMIM databases. The drug targets and bronchial asthma targets were intersected and imported String database was used to establish PPI network. Cytoscape 3.9.1 software was used to draw the network diagram of "Chinese materia medica-active components-intersection targets" and the core targets were screened. GO and KEGG enrichment analysis was performed on the core targets using DAVID database. A mouse model of asthma induced by ovalbumin was prepared. After the intervention of Kangmin Zhisou Granules, the pathological changes of mouse lung tissue were observed, and the contents of serum TNF-α, IL-6, IL-1 β were detected by ELISA.Results:Totally 240 active components and 1 364 potential targets were obtained from Kangmin Zhisou Granules. Tumor necrosis factor (TNF), interleukin-6 (IL-6), protein kinase B (AKT1), albumin (ALB), interleukin 1-beta (IL-1β) and other 11 core targets were obtained after screening. The results of GO enrichment analysis showed that the treatment of bronchial asthma by Kangmin Zhisou Granules mainly involved the positive regulation of protein phosphorylation, the regulation of inflammatory response, lipopolysaccharide response and other biological processes, as well as TNF, activated protein kinase (MAPK), interleukin-17 (IL-17) and other signaling pathways. Animal experiments confirmed that Kangmin Zhisou Granules could reduce the expression levels of TNF-α, IL-6 and IL-1β in serum ( P<0.05), and reduce the infiltration of inflammatory cells in the lung tissue of mice, thereby relieving asthma symptoms. Conclusion:Kangmin Zhisou Granules may exert anti-inflammatory effects by acting on TNF-α, IL-6, IL-1β and other targets to alleviate asthma symptoms.