Objective To observe the dynamic changes of TNF-α,IL-1β,IL-6,IL-10,and expressions of Notch-1,NF-κB mRNA and their protein levels in the brain tissue of septic mice and intervention effects of intrathecal injection of lentiviral vector of miR-34a gene for regulating Notch-1/NF-κB signaling pathway.Methods A total of 54 mice of clean grade were divided randomly (random number) into four groups,namely sham group (n =9),in which sham-operated laparotomy was performed;CLP group (n=15),in which the cecum ligation operation (CLP) was performed;NC group (n =15),in which intrathecal injection of lentivirus 5 μL (concentration 5 × 108 TU/mL),one time per day,for 3 days,then CLP was performed on the seventh day;intervention group (n =15),in wihch intrathecal injection of miR-34a lentivirus 5 μL (concentration 5 × 108 TU/mL),one time per day,for 3 days,then CLP was performed on the seventh day.The mice of four groups were sacrificed 24 h after modeling or operation.The changes of behavior of mice was observed and the neurological scores were assessed 24 h after CLP.The levels of TNF-α,IL-1β,IL-6 and IL-10 in the brain were measured by ELISA method.The mRNA expression and protein levels of Notch-1 and NF-κB in the brain tissue were measured by real-time PCR and Western blot method,respectively.Pathological changes of brain tissue were observed under light microscope.Results The neurological scores,the cerebral TNF-α,IL-6 levels,the mRNA expression and protein level of NF-κB (P＜0.01),and IL-1β levels (P ＜0.05) in CLP group 24 h after modeling were higher than those in sham group.The cerebral IL-10 level and the mRNA expression and protein level of Notch-1 (P ＜ 0.01) in CLP group 24 h after modeling were lower than those in sham group.The neurological scores,the cerebral TNF-α and IL-1β levels,and the protein level of NF-κB (P ＜0.01),and IL-6 (P ＜ 0.05) in NC group 24 h after modeling were higher than those in sham group.The cerebral IL-10 level,the mRNA expression and protein level of Notch1 in NC group 24 h after modeling were lower than those in sham group.There were no significant difference in neurological socres,IL-1β and IL-6 levels between intervention group and CLP group (P ＞ 0.05).The IL-10 level (P ＜ 0.05) and the mRNA expression (P ＜ 0.05) and protein level (P ＜ 0.01) of Notch-1 in intervention group 24 h after modeling were lower than those in CLP group.There were no significant difference in biomarkers between NC group and CLP group (P ＞ 0.05).The degree of cerebral damage found under light microscope was also ameliorated in intervention group compared with CLP group 24 h after modeling.Conclusions The effects of miR-34a via regulating Notch-1/NF-κB signaling pathway on brain function exerts cerebral protective effects in septic mice.

Objective To prepare a novel monoclonal antibody (mAb) that specifically against Mycobacterium tuberculosis culture filtrate protein 10 (CFP-10).Methods The BALB/c mice were immunized by a peptide with 14 amino acids (aa residues 53 to 66) of CFP-10,and then the splenocytes of mice were fused with myeloma cell line SP2/0.The resultant fused cells were subjected to screening culture,enzyme linked immunosorbent assay (ELISA) assay and subcloning by limited dilution to establish hybridoma cell lines of stable secreting anti-the peptide of CFP-10 antibody.The antibody was purified,and its isotypes were analyzed.Then,the antibody was further evaluated by Western blotting,immunoprecipitation and ELISA in 38 culture supernatant samples of Mycobacterium tuberculosis,20 culture supernatant samples of non-Mycobacterium tuberculosis,32 samples of tuberculous pleural effusion,24 samples of non-tuberculous pleural effusion,and 20 serum samples from healthy controls.Results The isotype of the mAb against the specific peptide of CFP-10 was an IgG1 with κ chain,and it was applicable for Western blotting and immunoprecipitation analysis.ELISA quantitative test showed that the sensitivity and specificity for diagnosis of Mycobacterium tuberculosis were 78.6％ (55/70) and 92.2％ (59/64),respectively.Conclusion The mAb generated against the specific peptide of CFP-10 is high in sensitivity and specificity,and it might be used in the early diagnosis of tuberculosis.

Objective To investigate the anti-inflammatory effect of C1q/ tumor necrosis factor (TNF)-related protein 9 (CTRP9) in RAW264.7 mouse macrophage cells treated with oxidized low density lipoprotein (oxLDL),and to explore its mechanism.Methods RAW264.7 mouse macrophage cells were divided into three groups:the control group,the oxLDL group (treated with oxLDl) and the gCTRP9-oxLDL group (pretreated with recombinant globular domain of CTRP9 and stimulated by oxLDL).Foam cells were detected by oil red O staining.Western blot was used to detect the expressions of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein 1 (MCP-1).In addition,the expression levels of NF-κB p65 in cytoplasm and nucleus proteins extraction were both determined.Results The relative levels of MCP-1 and NF-κB were increased in the oxLDL group as compared with the control group (1.66±0.09 vs.1.03±0.10,0.52±0.11 vs.1.03±0.06,both P＜0.05).The expression levels of TNF-α and MCP-1 were decreased in gCTRP9+oxLDL group as compared with the oxLDL group (both P＜0.05).The expression level of NF κB p65 in nucleus 2 and 8 h after treatment was lower in the gCTRP9+oxLDL group than in the oxLDL group (1.03±0.06 vs.0.17±0.10,0.31±0.03,both P＜0.05).Conclusions oxLDL may induce the expressions of inflammatory cytokines of TNF α and MCP-1 in macrophage ceils.gCTRP9 pretreatment could reduce the oxLDL-induced pro inflammatory effect and nuclear factor κB translocation may be involved in this process,which suggests that gCTRP9 may play a protective role in anti inflammatory and anti-atherosclerosis.

BACKGROUND:Reducing the rate of abnormal fertilization is an effective approach to improving the efficacy of in vitro fertilization and reducing patients'financial strain.However,the current research on abnormal fertilization has focused on exploring the types of prokaryotic nuclei and their generation mechanisms,as well as analyzing embryos formed by abnormal fertilization,chromosomal ploidy and utilization value.There is a lack of clinical prediction models for abnormal fertilization based on retrospective studies. OBJECTIVE:To construct a nomogram model to predict abnormal female factors in in vitro fertilization. METHODS:A total of 5 075 patients undergoing treatment for conventional in vitro fertilization at Nanxishan Hospital of Guangxi Zhuang Autonomous Region from March 2017 to March 2022 were retrospectively analyzed.The male confounders were calibrated on a 1:1 propensity score with a match tolerance of 0.02,and 1 672 cases were successfully matched.According to the Vienna Consensus,patients with≥60%normal fertilization capacity were included in the normal fertilization group(n=836)and those with<60%normal fertilization capacity were included in the abnormal fertilization group(n=836).The model and validation groups were obtained by random sampling at a ratio of 7:3.Factors related to the occurrence of abnormal fertilization following conventional in vitro fertilization in the model group were screened using univariate analysis and the best matching factors were selected using the Least Absolute Shrinkage and Selection Operator(LASSO)and included in a multifactorial forward stepwise Logistic regression to identify their independent influencing factors and plot a nomogram.Finally,the prediction model was validated for discrimination,accuracy and clinical application efficacy using receiver operating characteristic curves,calibration curves,clinical decision curves and clinical impact curves. RESULTS AND CONCLUSION:The univariate analysis indicated the factors influencing the occurrence of abnormal fertilization were age,controlled ovarian hyperstimulation protocol,number of assisted pregnancies,years of infertility,infertility factors,anti-mullerian hormone,sinus follicle count,basal luteinizing hormone,luteinizing hormone concentration on the human chorionic gonadotropin day,and estradiol level on human chorionic gonadotropin injection day(P<0.05).LASSO regression further identified the best matching factors,including age,microstimulation protocol,number of assisted pregnancies,years of infertility,anti-mullerian hormone,luteinizing hormone level on human chorionic gonadotropin injection day,and estradiol level on human chorionic gonadotropin injection day(P<0.05).Multifactorial forward stepwise Logistic regression results showed that age,microstimulation protocol,number of assisted conceptions,years of infertility,anti-mullerian hormone,and estradiol level on human chorionic gonadotropin injection day were independent influencing factors for the occurrence of abnormal fertilization following conventional in vitro fertilization.The receiver operating characteristic curves showed an area under the curve of 0.761(0.746,0.777)for the model group and 0.767(0.733,0.801)for the validation group,indicating that the model has good discrimination.The mean absolute error of the calibration curve was 0.044,and the Hosmer-Lemeshow test indicated that there was no significant difference between the predicted probability of abnormal fertilization and the actual probability of abnormal fertilization(P>0.05),indicating the prediction model has good consistency and accuracy.The clinical decision curves and clinical impact curves showed that the model and validation groups had the maximum net clinical benefit at valve probability values of 0.00-0.52 and 0.00-0.48,respectively,and there was a good clinical application efficacy in this valve probability range.To conclude,the nomogram model has good discrimination and accuracy as well as clinical application efficacy for predicting the occurrence of abnormal fertilization in women undergoing conventional in vitro fertilization based on age,microstimulation protocol,number of assisted conceptions,years of infertility,anti-mullerian hormone,and estradiol level on human chorionic gonadotropin injection day.