Objective To investigate the effect of drug-containing serum from Liuwei Dihuang Pill on Wnt/β-catenin pathway,migration and invasion of trophoblast cells induced by high glucose.Methods The drug-containing serum of Liuwei Dihuang Pill was prepared.Human chorionic trophoblast HTR-8/Svneo cells were cultured in vitro and divided into the control group(5 mmol/L glucose+blank serum),the high glucose group(30 mmol/L glucose+blank serum),the low-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 0.45 g/mL),the medium-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 0.90 g/mL)and the high-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 1.35 g/mL).The mRNA expressions of Wnt1,β-catenin and Cyclin D1 in HTR-8/Svneo cells were detected by real-time fluorescence quantitative PCR(qRT-PCR).The protein expressions of Wnt1,β-catenin,phosphorylated glycogen synthesis kinase-3β(p-GSK-3β)and Cyclin D1 in HTR-8/Svneo cells were detected by Western blot assay.The migration and invasion of HTR-8/Svneo cells were detected by Transwell method.Results Compared with the control group,the protein and mRNA expression levels of Wnt1,β-catenin and Cyclin D1,the numbers of migration cells and invasive cells in HTR-8/Svneo cells were decreased in the high glucose group(P<0.05).Compared with the high glucose group,the protein and mRNA expression levels of Wnt1,β-catenin and Cyclin D1,the numbers of migration cells and invasive cells were significantly increased in the low dose,the medium dose and the high dose groups of the HTR-8/Svneo cells(P<0.05).In addition,with the increase of the drug containing serum dose of Liuwei Dihuang Pill,the above indexes in HTR-8/Svneo cells increased successively(P<0.05).Conclusion The drug containing serum of Liuwei Dihuang Pil can activate Wnt/β-catenin pathway in HTR-8/Svneo cells induced by high glucose,and promote the migration and invasion of HTR-8/Svneo cells.

Objective To investigate the effect of drug-containing serum from Liuwei Dihuang Pill on Wnt/β-catenin pathway,migration and invasion of trophoblast cells induced by high glucose.Methods The drug-containing serum of Liuwei Dihuang Pill was prepared.Human chorionic trophoblast HTR-8/Svneo cells were cultured in vitro and divided into the control group(5 mmol/L glucose+blank serum),the high glucose group(30 mmol/L glucose+blank serum),the low-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 0.45 g/mL),the medium-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 0.90 g/mL)and the high-dose group(30 mmol/L glucose,serum medicine solution of Rehmannia Six Formula at 1.35 g/mL).The mRNA expressions of Wnt1,β-catenin and Cyclin D1 in HTR-8/Svneo cells were detected by real-time fluorescence quantitative PCR(qRT-PCR).The protein expressions of Wnt1,β-catenin,phosphorylated glycogen synthesis kinase-3β(p-GSK-3β)and Cyclin D1 in HTR-8/Svneo cells were detected by Western blot assay.The migration and invasion of HTR-8/Svneo cells were detected by Transwell method.Results Compared with the control group,the protein and mRNA expression levels of Wnt1,β-catenin and Cyclin D1,the numbers of migration cells and invasive cells in HTR-8/Svneo cells were decreased in the high glucose group(P<0.05).Compared with the high glucose group,the protein and mRNA expression levels of Wnt1,β-catenin and Cyclin D1,the numbers of migration cells and invasive cells were significantly increased in the low dose,the medium dose and the high dose groups of the HTR-8/Svneo cells(P<0.05).In addition,with the increase of the drug containing serum dose of Liuwei Dihuang Pill,the above indexes in HTR-8/Svneo cells increased successively(P<0.05).Conclusion The drug containing serum of Liuwei Dihuang Pil can activate Wnt/β-catenin pathway in HTR-8/Svneo cells induced by high glucose,and promote the migration and invasion of HTR-8/Svneo cells.

Objective:To analyze the drug-resistant gene loci of Mycoplasma pneumoniae (MP) using metagenomic next-generation sequencing (mNGS). Methods:From November 2022 to October 2023, 697 clinical samples (including sputum, alveolar lavage fluid and blood) of 686 children with Mycoplasma pneumoniae positive detected by mNGS were retrospectively analyzed. Samples were divided into intensive care unit (ICU) group and non-ICU group, Chi-square test was used to compare groups, and Mann-Kendall trend test was used to analyze the change trend of the detection rate of drug resistance gene loci over time. Results:Of the 697 samples, 164 were from the ICU group and 533 were from the non-ICU group. The detection rate of Mycoplasma pneumoniae resistance gene was 44.3% (309/697), and all detected drug-resistant gene loci of MP were A2063G. The detection rate of Mycoplasma pneumoniae in ICU group was 50.0% (82/164), and the detection rates of Mycoplasma pneumoniae resistance gene loci in sputum, alveolus lavage fluid and blood samples were 75.0% (18/24) and 48.4% (62/128), respectively. The detection rate in sputum was higher than alveolus lavage fluid samples ( χ2=5.72, P=0.017). The detection rate of Mycoplasma pneumoniae in non-ICU group was 42.6% (227/533), the detection rate of Mycoplasma pneumoniae resistance gene loci in sputum and alveolar lavage fluid was 40.0% (16/40), 44.3% (201/454), and no detection rate in blood samples (0/12). There was no significant difference in the detection rate of alveolar lavage fluid and sputum ( χ2=0.27, P=0.602). From November 2022 to October 2023, the detection rate of submitted samples showed an increasing trend month by month (overall: Z=3.99, ICU inspection group: Z=2.93, non-ICU group: Z=3.01, all P<0.01). Among the bacteria commonly detected with Mycoplasma pneumoniae, Streptococcus pneumoniae accounted for the highest proportion, the detection rate was 15.5% (108/697), and Epstein-Barr virus accounted for the highest proportion of 17.6% (123/697). Conclusions:From November 2022 to October 2023, the detection rate of Mycoplasma pneumoniae drug resistance gene loci showed an increasing trend. The detection rate of drug resistance gene loci in sputum samples of ICU group was higher than alveolus lavage fluid. No new drug resistance site were detected.

OBJECTIVE To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS) method for determination and comparison of 26 components in different parts of two base plants of Shiliang tea(Chimonanthus salicifolius S.Y.Hu and Chimonanthus zhejiangensis M.C.Liu), and screen quality markers of different parts. METHODS The UHPLC method was performed on an Agilent RRHD Eclipse Plus C18 (2.1 mm×50 mm, 1.8 μm) column with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.3 mL·min−1, the column temperature was 35 ℃, and the injection volume was 0.5 μL; the multiple reaction monitoring mode was employed for the quantification of 26 components with electrospray ionization(ESI) source polarity in negative and positive mode. RESULTS Good linear relationship(r >0.999) were observed in the test ranges for 26 compounds, and the average recovery was 88.5%−111.7% with RSD was 3.4%−9.8%. There was no significant difference between the two base plants of Shiliang tea, and all of these samples were divided into two categories by hierarchical cluster analysis. The main components in leaves was flavonoids, among them, the content of kaempferol 3-O-rutinoside was the highest, reaching 12.902 mg.g−1; the main components in stems and roots was coumarins, and the content of alkaloids in roots was higher, relatively; 7 quality markers of difference were screened by OPLS-DA, which were kaempferol 3-O-rutinoside, chimonanthine, rutin, fraxetin, calycanthoside, scopolin, neochlorogenic acid. CONCLUSION These study elucidates the differences of chemical components in the different parts of two base plants of Shiliang tea, which providing basis for the research of pharmacodynamic substances and references for the comprehensive utilization of Chimonanthus salicifolius S.Y. Hu and Chimonanthus zhejiangensis M.C.Liu resources.

Objective To explore the effects of Kaixin Jieyu Pill(KJP)on peroxisome proliferator-activated receptor alpha(PPARα),serum and hippocampal tissue fibroblast growth factor 21(FGF21)and hippocampal tissue fibroblast growth factor receptor 1(FGFR1)in rats with depression model.Methods A rat chronic stress model was prepared using the chronic unpredictable mild stimulus method(CUMS)to observe behavioral changes in depression.Hippocampal FGF21,FGFR1 and liver PPARα protein and mRNA expression levels were measured in hippocampal tissues by western blot and real-time fluorescence quantification(RT-qPCT).Results The expression levels of PPARα,FGF21 and FGFR1's protein and their mRNA were dramatically decreased in the CUMS model rats compared with the normal group(P<0.001).KJP could modify the depressive-like behaviors of rats(P<0.001),upregulate the protein expression level of PPARα,FGF21,and FGFR1(P<0.05,P<0.01,P<0.01);markedly modify the mRNA expression levels of PPARα,FGF21,and FGFR1(P<0.001,P<0.01,P<0.01).Conclusion PPARα/FGF21/FGFR1 pathway may participate in the pathogenesis of depression;upregulation of PPARα/FGF21/FGFR1 pathway may be related to the antidepressant mechanism of KJP.

Objective To evaluate the dynamic efficacy of Invigorating Qi and Dissipating Depression Herbs in the treatment of depressive disorders and its effect on recurrence rate and suicidal thoughts.Methods The prospective cohort study was used to divide the patients into three cohorts according to the different exposure factors(treatment regimens):Western medicine group,integrated traditional Chinese and Western medicine group,and Invigorating Qi and Dissipating Depression Herbs group,and followed up for 2 years,and the treatment effects of each cohort at half a year,one year,one and a half years,and two years were observed respectively,and the evaluation indicators were Montgomery Depression Scale(MADRS),Hamilton Anxiety Scale(HAMA)and Social Deficit Screening Scale(SDSS)reduction rates,and endpoint events(relapse,suicidal ideation)incidence;By comparing the efficacy and endpoint event rate of the three cohorts,the effect of Invigorating Qi and Dissipating Depression Herbs on improving depressive disorders was evaluated.Results A total of 409 patients(67 in the Western medicine group,131 in the integrated traditional Chinese and Western medicine group,and 211 cases in the Invigorating Qi and Dissipating Depression Herbs group)were enrolled,and the rate of score reduction after treatment with MADRS,HAMA and SDSS increased compared with the pre-treatment treatment with the extension of the drug taking time.The difference between the MADRS and SDSS reduction rates in the Invigorating Qi and Dissipating Depression Herbs group on taking the drug for half a year and the SDSS reduction rate on one year of taking the drug were statistically significant compared with the Western medicine group,and there was no significant difference compared with the integrated Chinese and Western medicine group.On one and a half years and two years of medication,the integrated Chinese and Western medicine group had more advantages in MADRS reduction rate than the Invigorating Qi and Dissipating Depression Herbs group.The recurrence rates of depression in the three groups within 2 years were 20.9％,30.5％and 36.0％,and the difference was not statistically significant,and the length of taking medication was used as a protective factor by logistic regression analysis.The incidence rates of suicidal thoughts were 44.8％,19.1％and 17.5％,and the differences were statistically significant,and the exposure factors and income level were the protective factors by logistic regression analysis.Conclusion Invigorating Qi and Dissipating Depression Herbs can significantly improve the depressive state of patients,promote the recovery of social function,reduce the occurrence of long-term suicidal ideation,and appropriately extend the time of taking medicine can increase the long-term efficacy and reduce recurrence.

Objective:To study the correlation between non-contact anterior cruciate ligament (ACL) injury and functional ankle instability (FAI) in young patients.Methods:A retrospective analysis was conducted of the 102 patients with non-contact ACL injury[61 males and 41 females, with an age of (31.9±6.1) years and a Tegner activity score of (6.1±1.9) points] who had been treated at Department of Orthopedics, Sun Yat-sen Memorial Hospital from January 2017 to March 2020 (injury group). Another 102 citizens without ACL injury from Guangzhou [56 males and 46 females, with an age of (30.3±7.2) years and a Tegner activity score of (6.0±2.1) points] were recruited as a control group. The Cumberland ankle instability tool (CAIT) and the Ankle Joint Functional Assessment Tool (AJFAT) were used to assess whether the subjects had self-conscious FAI or not. A correlation analysis was conducted using the data collected.Results:The 2 groups were comparable because there were no significant differences between them in general data ( P>0.05). By the CAIT score, the incidence of FAI in the injury group [52.9% (54/102)] was significantly higher than that in the control group [32.4% (33/102)] ( P<0.05); by the AJFAT score, the incidence of FAI in the injury group [59.8% (61/102) ] was significantly higher than that in the control group [39.2% (40/102)] ( P<0.05). Pearson correlation analysis showed that diagnoses of FAI by CAIT and by AJFAT were respectively correlated with ACL injury ( r=-0.159, P=0.023; r=-0.215, P=0.002). Conclusions:The incidence of FAI may be high in patients with ACL injury and there is a correlation between FAI and ACL injury.

Objective To analyze the changes of Survivin,MSH6 and MSH2 expression in colorectal cancer and explore their relationship with clinical and pathological parameters.Methods 197 cases of colorectal adenocarcinoma and 20 cases of inflammatory intestinal mucosa were detected by immunohistochemistry with Survivin,MSH6 and MSH2,and the correlation between Survivin,MSH6 and MSH2 expression was analyzed.Results In the control group of 20 cases with inflammatory non-tumor intestinal mucosa,the positive expression rate of MSH2,MSH6,Survivin were 95％,95％,10％,respectively.While the positive expression rate of MSH2,MSH6,Survivin were 88.3％,74.1％ and 84.3％ in 197 cases of colorectal cancer.Survivin positive expression rate in colorectal cancer group was significantly higher than that in inflammatory control group (P ＜ 0.05).Survivin expression was correlated with invasion depth and lymph node metastasis in colorectal cancer tissue (P ＜ 0.05),but not with gender,age,tumor size,gross type or degree of differentiation (P ＞ 0.05).MSH2 expression was correlated with tumor size and lymph node metastasis (P ＜ 0.05),but not with gender,age,gross type,depth of infiltration and degree of differentiation (P ＞ 0.05).MSH6 expression was related to gender and lymph node metastasis (P ＜ 0.05),but not to age,tumor size,gross type,infiltration depth and differentiation (P ＞ 0.05).Colorectal cancer tissues showed positive correlation between MSH6 and MSH2 (r =0.326,P ＜ 0.01),positive correlation between MSH2 and Survivin positive expression (r =0.277,P ＜ 0.01),and positive correlation between MSH6 and Survivin positive expression (r =0.435,P ＜ 0.01).Conclusions The positive expression rate of Survivin in colorectal cancer is high.MSH2,MSH6 and Survivin in colorectal cancer play an important role in the development and progression of colorectal cancer,and can provide evidence for the detection of these three factors,including metastasis risk,prognosis assessment and clinical treatment.In particular,Survivin gene may provide evidence for gene therapy.

Objective: To investigate the expression of serum Neurensin 2 (NRSN2) in patients with nasopharyngeal carcinoma (NPC), and to analyze its relationships with epstein-barr virus (EBV) antibody and EBV-DNA. Methods: 120 patients with NPC admitted to our hospital from April 2016 to May 2018 were selected as the study group, and 56 healthy people in the physical examination center were selected as the control group. Enzyme linked immunosorbent assay (ELISA) was used to detect the expressions of serum viral capsid antigen-IgA (VCA-IgA), nuclear related tumor antigen-IgA (EBNA-IgA) and early antigen-IgG (EA-IgG). The expression of serum NRSN2 and EBV-DNA were detected by fluorescence quantitative polymerase chain reaction (qPCR). Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of serum NRSN2 level for NPC. Spearman was used to analyze the relationship between serum NRSN2 and EBV antibody with EBV-DNA in nasopharyngeal carcinoma, multivariate logistic regression analysis was used to analyze the risk factors of NPC. Results: The positive rates of serum VCA-IgA, EBNA-IgA and EA-IgG in the study group were significantly higher than those in control group (P<0.05); the level of serum NRSN2 and the positive expression rate of EBV-DNA in the study group was significantly higher than that in the control group (P<0.05); ROC curves showed that the area under curve (AUC) of serum NRSN2 level in diagnosing NPC disease was 0.759, with sensitivity 63.33%, specificity 80.36%; there were significant positive correlations between serum NRSN2 with VCA-IgA, EBNA-IgA, EA-IgG and EBV-DNA in NPC patients (P<0.05); multivariate regression analysis showed that VCA-IgA, EBNA-IgA, EA-IgG, EBV-DNA and NRSN2 expressions were supportive predictive value for diagnostic NPC (P<0.05). Conclusions Up-regulation of serum NRSN2 levels in NPC patients is positively correlated with serum VCA-IgA, EBNA-IgA, EA-IgG and EBV-DNA levels, which may provide a reference for the prediction, diagnosis and treatment of NPC.

Objective:To investigate the tumor-specific neoantigen for primary plasma cell leukemia (PCL) using gene sequencing technology combined with bioinformatic analysis. Methods: Peripheral blood samples of one patient with primary PCL during relapse and remission periods were collected. HLA molecular typing was performed using polymerase chain reaction with sequencing-based typing; whole-exome and transcriptome were sequenced by next-generation sequencing method; and bioinformatics software NetMHCpan was used to predict neoantigens. Results: Six tumor-specific missense mutations were found in the patient's peripheral blood during relapse period, located in genes FRG1, MLL3, SVIL, MYOM1, ZDHHC11 and RFPL4A.Considering patient's HLA sub-types, 43 neoantigens were predicted via bioinformatics. Considering that FRG1 and MLL3 had relatively high gene expression levels, 20 neoantigens derived from mutations of the two genes were preferentially selected, among which four neoantigens had high affinity with the patient's HLA molecules and thus had potential clinical application value. Conclusion: The study has completed a tumor neoantigen screen and prediction for primary PCL. This practice demonstrates that predicting neoantigen based on tumor-specific somatic mutation is feasible for primary PCL.