Serum levels of soluble programmed death-1 and soluble programmed death-ligand 1 in chronic hepatitis B patients with clinical cure and their clinical features / 临床肝胆病杂志

Objective To investigate the serum levels of soluble programmed death-1 (sPD-1) and soluble programmed death-ligand 1 (sPD-L1) in chronic hepatitis B (CHB) patients with clinical cure, the correlation between programmed death-1 (PD-1) and lymphocytes by flow cytometry, and the recovery of hepatitis B virus (HBV)-specific immunity. Methods A total of 26 CHB patients with clinical cure, 26 treatment-naïve CHB patients, and 26 healthy controls who were diagnosed at the outpatient service of Peking University First Hospital from January to May of 2022 were enrolled, and related clinical data and peripheral blood samples were collected. ELISA was used to measure the serum levels of sPD-1 and sPD-L1, and flow cytometry was used to measure the expression of PD-1 in peripheral blood lymphocytes. CHB patients with clinical cure were compared with the treatment-naïve CHB patients and the healthy controls. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between three groups, and the chi-square test was used for comparison of categorical data between groups. The Pearson correlation analysis or the Spearman correlation analysis was used to investigate the correlation between two continuous variables. Results For the 26 CHB patients with clinical cure, the mean time of antiviral therapy was 8.33 years, with entecavir as the antiviral drug. The CHB patients with clinical cure had significantly higher levels of sPD-1 and sPD-L1 than the healthy controls ( P 0.05). Conclusion The serum levels of sPD-1 and sPD-L1 in treatment-naïve CHB patients are mainly associated with exhausted CD8 + T cells in peripheral blood, while there is no significant correlation between serum sPD-1/sPD-L1 and exhausted CD8 + T cells in peripheral blood in CHB patients with clinical cure.

Diagnostic value of novel hepatic fibrosis markers in assessing cirrhosis in patients with chronic hepatitis C / 中华肝脏病杂志

Objective: To investigate the efficacy of chitinase-3-like protein 1 (CHI3L1) and Golgi protein 73 (GP73) in the diagnosis of cirrhosis and the dynamic changes of CHI3L1 and GP73 after HCV clearance in patients with chronic hepatitis C (CHC) treated with direct-acting antiviral drugs (DAAs). The comparison of continuous variables of normal distribution were statistically analyzed by ANOVA and t-test. The comparison of continuous variables of non-normal distribution were statistically analyzed by rank sum test. The categorical variables were statistically analyzed by Fisher's exact test and χ(2) test. Correlation analysis was performed using Spearman correlation analysis. Methods: Data of 105 patients with CHC diagnosed from January 2017 to December 2019 were collected. The receiver operating characteristic curve (ROC curve) was plotted to study the efficacy of serum CHI3L1 and GP73 for the diagnosis of cirrhosis. Friedman test was used to compare CHI3L1 and GP73 change characteristics. Results: The areas under the ROC curve for CHI3L1 and GP73 in the diagnosis of cirrhosis at baseline were 0.939 and 0.839, respectively. Serum levels of CHI3L1 and GP73 in the DAAs group decreased significantly at the end of treatment compared with baseline [123.79 (60.25, 178.80) ng/ml vs. 118.20 (47.68, 151.36) ng/ml, P = 0.001; 105.73 (85.05, 130.69) ng/ml vs. 95.52 (69.52, 118.97) ng/ml, P = 0.001]. Serum CHI3L1 and GP73 in the pegylated interferon combined with ribavirin (PR) group were significantly lower at the end of 24 weeks of treatment than the baseline [89.15 (39.15, 149.74) ng/ml vs. 69.98 (20.52, 71.96) ng/ml, P < 0.05; 85.07 (60.07, 121) ng/ml vs. 54.17 (29.17, 78.65) ng/ml, P < 0.05]. Conclusion: CHI3L1 and GP73 are sensitive serological markers that can be used to monitor the fibrosis prognosis in CHC patients during treatment and after obtaining a sustained virological response. Serum CHI3L1 and GP73 levels in the DAAs group decreased earlier than those in the PR group, and the serum CHI3L1 levels in the untreated group increased compared with the baseline at about two years of follow-up.

Influencing factors for direct-acting antiviral therapy failure in treatment of hepatitis C / 临床肝胆病杂志

Objective To investigate the influencing factors for direct-acting antiviral agent (DAA) therapy failure in the treatment of hepatitis C by comparing baseline clinical data and resistance-associated substitution (RAS) in sequencing data between the patients with HCV RNA reactivation after DAA therapy and the patients with successful DAA treatment. Methods A total of 13 patients from multiple centers who failed DAA therapy from November 2019 to October 2021 were enrolled as treatment failure group, and sequencing was performed for their positive serum samples. A total of 51 patients with successful DAA treatment were enrolled as control group, and baseline clinical data and sequencing results were compared between the treatment failure group and the control group. The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups, and the chi-square test was used for comparison of categorical data between groups; univariate and multivariate logistic regression analyses were performed to calculate odds ratio ( OR ) and investigate the influencing factors for treatment failure. Results All 12 patients with complete treatment data experienced recurrence within 1 year after the end of medication. The male patients with treatment failure had significantly higher baseline total bilirubin, direct bilirubin, and creatinine than their female counterparts ( Z =-2.517, -2.440, and -2.132, P =0.010, 0.010, and 0.038), and the patients with an age of ≤55 years ( OR =5.152, 95% confidence interval [ CI ]: 1.116-23.790, P =0.036) or genotype 3b ( OR =9.726, 95% CI : 1.325-71.398, P =0.025) had a higher probability of treatment failure. There were differences in the incidence rates of major RAS mutations on three gene fragments between the treatment failure group and the treatment success group, and the common RAS mutations detected in the treatment failure group were not detected in the treatment success group. Conclusion Age, genotype, and RAS in serum virus gene sequence are influencing factors for DAA treatment failure.

Erianin Inhibits Proliferation of Bladder Cancer 5637 Cells Through Akt / 中国实验方剂学杂志

ObjectiveTo investigate the role of protein kinase B （Akt） overexpression in the inhibition of human bladder cancer 5637 cell proliferation by erianin and related mechanisms. MethodThe 5637 cells stably over-expressing Akt were induced using the lentivirus vector. The 5637 cells infected with the empty vector were classified into blank group. Then the Akt group， empty vector combined with erianin （62.5 μg·L-1） group， and Akt combined with erianin （62.5 μg·L-1） group were set up. The cell viability was detected by cell counting kit-8 （CCK-8） assay， and the clone formation of 5637 cells in each group was determined in the clone formation experiment. The cell cycle distribution was detected by flow cytometry. Western blot was used to assay the protein expression levels of phosphorylated （p）-Akt， Akt， p21. The glycolysis of 5637 cells was determined in glucose uptake and lactate secretion assays. ResultCompared with the blank group， erianin inhibited the proliferation of bladder cancer 5637 cells （P<0.05）. Overexpression of Akt partially reversed the inhibitory effect of erianin on the proliferation of bladder cancer 5637 cells （P<0.05）. Clone formation assay showed that erianin inhibited the clone formation of bladder cancer 5637 cells （P<0.05）， which was partially reversed by the overexpressed Akt （P<0.05）. As revealed by comparison with the blank group， erianin arrested the bladder cancer 5637 cells in G1 phase （P<0.05）， which was also reversed by the overexpressed Akt （P<0.05）. Western bolt showed that erianin promoted the expression of p21 but suppressed the expression of p-Akt and Akt （P<0.05）. By contrast， the overexpression of Akt down-regulated the elevated p21 protein expression induced by erianin （P<0.05）. Compared with the blank group， erianin inhibited the glucose uptake and lactate secretion of bladder cancer 5637 cells （P<0.05）. Overexpression of Akt weakened the inhibitory effect of erianin against the glycolysis of 5637 cells （P<0.05）. ConclusionErianin is able to inhibit the proliferation of bladder cancer 5637 cells， promote the expression of p21， and inhibit the expression of p-Akt. Overexpressed Akt reduces the inhibitory effect of erianin on the proliferation of bladder cancer 5637 cells， suggesting that Akt plays an important role in the inhibition of 5637 cell proliferation by erianin， which has provided a new target for the application of erianin in the treatment of bladder cancer.

Effect of Gupi Xiaoji Decoction on Pyroptosis of HepG2.2.15 Cells Based on Network Pharmacology and Molecular Docking / 中国实验方剂学杂志

ObjectiveTo screen the active antitumor components of Gupi Xiaoji decoction by network pharmacology and molecular docking based on the pyroptosis mediated by cysteinyl aspartate-specific protease 1 （Caspase-1） and explore its molecular mechanism in intervening in the pyroptosis of HepG2.2.15 cells through in vitro experiments. MethodThe compounds and targets of Gupi Xiaoji decoction were screened out by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） to obtain the corresponding gene symbols. The targets of Caspase-1 were collected from GeneCards，online mendelian inheritance in man（OMIM），PharmGKB，and TTD，and the compound-gene target regulatory network was constructed by Cytoscape. The protein-protein interaction（PPI） network was established and analyzed by STRING. The mechanism of the effective components of Gupi Xiaoji decoction on Caspase-1 was predicted by gene ontology（GO） functional enrichment and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analyses. The molecular docking was verified with AutoDock Vina. The plasma medicated with Gupi Xiaoji Decoction was prepared and HepG2.2.15 cells were cultured in vitro. HepG2.2.15 cells were divided into a blank plasma group，a VX-765 group，a VX-765+medicated plasma group， and a medicated plasma group. After 48 hours of intervention with 15% medicated plasma， the expression and distribution of gasdermin D-N （GSDMD-N） on the surface of the cell membrane were detected by immunofluorescence staining. The release of lactic dehydrogenase （LDH）， interleukin（IL）-1β，and IL-18 in the cell supernatant was measured by enzyme-linked immunosorbent assay（ELISA） kits. The expression of Caspase-1 and GSDMD-N was measured by Western blot. ResultThe mitogen-activated protein kinase 14 （MAPK14），MAPK1，protein kinase B1 （Akt1）， MAPK8， V-Jun sarcoma virus oncogene homolog （JUN）， and TP53 screened by network pharmacology were the main targets. The compounds 7-hydroxy-5，8-dimethoxy-2-phenyl-chromone，wogonin，rhamnazin，moslosooflavone，isorhamnetin，7-O-methylisomucronulatol，formononetin，calycosin，luteolin，quercetin，kaempferol，β-sitosterol，and baicalein screened by network pharmacology were the main active components of Gupi Xiaoji decoction. Go enrichment analysis showed that multiple biological processes were involved， including responses to oxidative stress and metal ions，ubiquitin-like protein ligase binding，and phosphatase binding. KEGG pathway enrichment analysis showed MAPK pathway，nuclear factor（NF）-κB pathway，p53 pathway， and hypoxia-inducible factor-1（HIF-1） pathway were involved. Molecular docking showed that the targets had good binding with the components. In vitro experiments displayed that compared with the blank plasma group，the VX-765 group showed weakened GSDMD-N fluorescence signal，reduced release of LDH，IL-1β，and IL-18，and declining expression of Caspase-1 and GSDMD-N（P<0.01）， and the medicated plasma group showed increased GSDMD-N fluorescence signal， increased release of LDH，IL-1β，and IL-18，and up-regulated expression of Caspase-1 and GSDMD-N（P<0.01）. ConclusionGupi Xiaoji Decoction can induce the pyroptosis of HepG2.2.15 cells by regulating Caspase-1 through multiple targets and multiple pathways.

Virological response to direct-acting antiviral therapy and changes in liver fibrosis indices in chronic hepatitis C patients with different alanine aminotransferase and aspartate aminotransferase levels in a real-world setting / 临床肝胆病杂志

ObjectiveTo investigate the virologic response to direct-acting antiviral (DAA) therapy and the changes in liver stiffness measurement (LSM), fibrosis-4 (FIB-4), and aspartate aminotransferase-to-platelet ratio index (APRI) after treatment in chronic hepatitis C (CHC) patients with different alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels at baseline in a real-world setting. MethodsCHC patients who attended the outpatient service of Department of Infectious Diseases, Peking University First Hospital, from December 2017 to May 2020 were enrolled, and virologic response rate was calculated. The Wilcoxon rank-sum test was used to compare LSM, FIB-4, and APRI between groups at baseline and at 12 weeks after treatment, and the chi-square test was used for comparison of categorical data between groups. ResultsA total of 48 CHC patients were enrolled, among whom 33.3% had abnormal ALT or AST at baseline. Among these patients, the virologic response rate was 85.4% at week 4 of treatment and 100% at the end of treatment and at 12, 24, and 48 weeks after treatment, and there were significant changes from baseline to 12 weeks after treatment in LSM ［6.1 (51-12.4) kPa vs 8.6 (5.7-16.9) kPa, Z=-1.676, P=0.043］ and APRI ［0.24(0.19-0.48) vs 0.42(0.23-1.17), Z=-2.050, P=0027］. From baseline to 12 weeks after treatment, the patients with abnormal ALT or AST at baseline had significant changes in LSM ［89(5.6-13.1) kPa vs 14.4(8.0-28.2) kPa, Z=-1.679, P=0.047］ and APRI ［0.44(0.25-0.50) vs 1.29(0.99-2.09), Z=-3.427, P=0.001］. ConclusionCHC patients achieve a high sustained virologic response rate after DAA therapy, and the patients with abnormal ALT or AST at baseline tend to have more significant improvements in LSM and APRI than those without such abnormality.

The predictive value of general movement assessment combined total bilirubin for motor development outcomes in infants with severe neonatal jaundice / 中国医师杂志

Objective:To study the predictive values of the general movements (GMs) assessment combined total bilirubin for motor development outcomes in infants with severe neonatal jaundice.Methods:From June of 2014 to June of 2019, infants with severe neonatal jaundice in Chenzhou First People′s Hospital were enrolled in the study. Inclusion criteria included , the serum total bilirubin was measured at the time of admission, corrected gestational age of 37 to 48 weeks. General assessment were carried out when the infant was stable. The patients were regularly followed-up until the age of 12months to evaluate the predictive values.Results:A total of 204 patients with severe neonatal jaundice were enrolled in the study, with mean serum total bilirubin value (485.4±109.6)μmol/L. They were divided into two groups according to the outcome of motor development. The total bilirubin value, the proportion of abnormal GMs and dangerous total bilirubin level in the abnormal group were significantly higher than those in the normal group (all P<0.05). 13 cases (6.4%) were normal in the torsion stage of GMs; 191 cases (93.6%) were abnormal, including 164 cases (85.9%) of poor repertoire (PR) and 27 cases (14.1%) of cramped-synchronized (CS). Abnormal GMs and total bilirubin were the risk factors of abnormal motor development ( OR=4.651, 1.017, P<0.05). The predictive values of abnormal GMs for abnormal motor development outcomes were as following: sensitivity 100%, specificity 8.4%, negative predictive value (NPV) 100%. The predictive values of CS for cerebral palsy were as following: sensitivity 63.2%, specificity 97.8%, NPV 96.0%. Receiver operating characteristic (ROC) curve showed that the area under the curve predicted by GMs and total bilirubin was 0.765 and 0.757, respectively. The area under the curve of motor dysplasia predicted by combining the two was 0.854. Conclusions:The evaluation of general movement assessment combined total bilirubin has certain clinical predictive value for the outcomes of motor development in infants with severe neonatal jaundice.

Effect of Jianpi Xiao' ai Prescription on Akt/mTOR Signaling Pathways in Colorectal Cells / 中国实验方剂学杂志

Objective::To investigate the effect of drug-containing serum of Jianpi Xiaoai prescription on protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathways in colorectal cells HCT116. Method::The HTC116 cells were treated by 15%concentration of drug-contained serum, and then the cell migration and invasion were detected by Transwell assay, the protein expression levels of Akt, phosphorylated protein kinase B (p-Akt), mTOR, phosphorylated mammalian target of rapamycin (p-mTOR), ribosomal protein S6 kinase, polypeptide1(S6K1), phosphorylated ribosomal protein S6 kinase, polypeptide1 (p-S6K1), 4E-binding protein1(4EBP1), and phosphorylated 4E-binding protein1(p-4EBP1) in HCT116 cells were detected by Western blot. The control group was treated by untreated serum (15%), and 10%fetal bovine serum(FBS). Result::As compared with the control group, the number of migration and invasion cells was significantly reduced in drug-contained serum group (P<0.01), the expression of Akt had no obvious decrease, p-Akt protein expression was significantly lowered in the drug-contained serum group (P<0.01), the expression of mTOR had no obvious decrease, but p-mTOR protein expression was significantly lowered in drug-contained serum group (P<0.01), the expression of S6K1 had no obvious decrease, but p-S6K1 protein expression was significantly lowered in the drug-contained serum group (P<0.01), the protein expression of 4EBP1 had no obvious decrease, but p-4EBP1 protein expression was significantly lowered in the drug-contained serum group (P<0.01). Conclusion::The anti-tumor mechanism and transfer of Jianpi Xiaoai prescription may be related to inhibiting the activation of Akt/mTOR signaling pathways in colorectal cancer.

Effect of Polyphyllin Ⅰ on Transcriptional Factor FOXQ1 and Epithelial-mesenchymal Transition in Colorectal Cancer Cell HCT116 / 中国实验方剂学杂志

Objective:To observe the effect of polyphyllin Ⅰ on the expressions of forkhead box Q1（FOXQ1）and epithelial-mesenchymal transition (EMT)-related factors, in order to explore the possible mechanism of polyphyllin Ⅰ in inhibiting the metastasis of colon cancer. Method:After the treatment with 1.25，2.50 μmol·L-1 polyphyllin Ⅰ on HCT116 cells, Western blot and Real-time PCR were used to detect the expressions of FOXQ1，E-cadherin，Vimentin protein and mRNA. Result:Compared with the blank group, relative expressions of FOXQ1 protein and mRNA in low-concentration polyphyllin Ⅰ group were decreased, while relative expression of E-cadherin mRNA was increased, the differences were not statistically significant, and relative expressions of Vimentin protein and mRNA in low-concentration polyphyllin Ⅰ group were decreased (P<0.05)， and relative expression of E-cadherin protein in low-concentration polyphyllin Ⅰ group was increased (P<0.01). Compared with the blank group, relative expressions of FOXQ1, Vimentin protein and mRNA in high-concentration polyphyllin Ⅰ group were decreased，while relative expressions of E-cadherin protein and mRNA were increased (P<0.05, P<0.01). Compared with low-concentration polyphyllin Ⅰ group, relative expressions of Vimentin protein and mRNA in high-concentration polyphyllin Ⅰ group were decreased, but the difference was not statistically significant, relative expressions of E-cadherin protein and mRNA in high-concentration polyphyllin Ⅰ group were increased, whereas relative expressions of FOXQ1 protein and mRNA were decreased (P<0.05, P<0.01). Conclusion:Mechanism of polyphyllin Ⅰ inhibiting the metastasis of colon cancer may be related to the decrease of FOXQ1 and Vimentin expressions, and the up-regulation of E-cadherin.

Effect of Jianpi Xiaoai Prescription on Activation of HFL1 into CAFs Induced by Exosomes in Colon Cancer / 中国实验方剂学杂志

Objective:To observe the effect of Jianpi Xiaoai prescription on the activation of normal human embryonic lung fibroblasts （HFL1） into tumor-associated fibroblasts （CAFs） induced by human colon cancer cells （HCT116） derived exosomes. Method:SD rats were gavaged with 13.1 g·kg-1 of Jianpi Xiaoai prescription to prepare drug-containing serum， and HCT116 cell exosomes-containing 10% exosomes-free serum and 20% Jianpi Xiaoai prescription drug serum were isolated by ultra-high speed centrifugation. The particle size distribution of exosomes were detected by Nanoparticle tracking analyzer （Zetaview）， and the exosomes' marker proteins apoptotic transfer gene 2 interaction protein X （Alix）， heat shock protein 70 （HSP70）， and tumor-susceptibility gene 101 （TSG101） were identified by Western blot， and the uptake of exosomes labeled with cell membrane staining kit （PKH67） by HFL1 was observed by fluorescence microscope. HFL1 cells were divided into six groups： the blank group， the transforming growth factor-β1 （TGF-β1） group， the TGF-β1 combined with HCT116 exosomes of 2 mg·L-1 group， the TGF-β1 combined with HCT116 exosomes of 4 mg·L-1 group， the TGF-β1 combined with Jianpi Xiaoai prescription exosomes of 2 mg·L-1 group， and the TGF-β1 combined with Jianpi Xiaoai prescription exosomes of 4 mg·L-1 group， and all groups were cultivated for 48 h. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to determine the protein and mRNA expressions of α-smooth muscle actin （α-SMA）. Result:The particle size distribution detected by Zetaview was mainly between 50-100 nm， and the exosomes were verified based on the expressions of marker proteins Alix， HSP70 and TSG101. After co-incubation of HFL1 cells with exosomes， a large number of exosomes were absorbed by HFL1 cells under fluorescence microscope. Compared with the blank control group， the protein and mRNA expressions of α-SMA in the TGF-β1 group and TGF-β1 combined with HCT116 exosome groups were increased （P<0.01）. Compared with the TGF-β1 combined with HCT116 exosome groups， the protein and mRNA expressions of α-SMA were decreased in the TGF-β1 combined with Jianpi Xiaoai prescription exosome groups （P<0.01）. Conclusion:Human colon cancer cell exosomes combined with TGF-β1 can induce the activation of HFL1 into CAFs， and Jianpi Xiaoai prescription can reduce the activation of HFL1 by affecting the expressions of α-SMA， thus antagonizing the lung metastasis of colon cancer.

The predictive value of writhing stage general movement assessment for motor development outcomes in infants with severe neonatal jaundice / 中国新生儿科杂志

Objective To study the predictive values of the general movements (GMs) assessment in writhing stage for motor development outcomes in infants with severe neonatal jaundice.Method From December of 2012 to December of 2017,infants with severe neonatal jaundice (serum bilirubin reaching the corresponding level of exchange transfusion according to the reference nomogram) in our hospital were enrolled in the study.Inclusion criteria included corrected gestational age of 37 to 48 weeks,serum bilirubin level below phototherapy intervention value after treatment and general and detailed assessment were carried out in writhing stage when the infant was stable.The patients were regularly followed-up until one-year-old to evaluate the predictive values.Result A total of 241 patients with severe neonatal jaundice were enrolled in the study,including 153 males (63.5％) and 88 females (36.5％),with gestational age between 35 and 42 weeks.The mean gestational age was (37.9± 1.8) weeks,the average birth weight was (3 057±480) g,and the mean serum bilirubin value was (458.9± 119.1) μmol/L.The general evaluation of the GMs was normal in 15 cases (6.2％),and abnormal in 226 cases (93.8％) with 217 cases (90.0％) were poor repertoire (PR) and 9 cases (3.7％) were cramped-synchronized (CS).The predictive values of abnormal GMs for abnormal motor development outcomes were as following:sensitivity 100％,specificity 7.6％,negative predictive value(NPV) 100％.The predictive values of CS for cerebral palsy were as following:sensitivity 22.2％,specificity 97.8％,NPV 94.0％.Detailed evaluation of 241 subjects showed that 13 items had statistically significant differences in the prediction of cerebral palsy (P＜0.05),and 18 items in the prediction of abnormal motor development (P＜0.05).Conclusion The CS pattern and detailed assessment of GMs in the writhing stage may be correlated with the outcomes of motor development in infants with severe neonatal jaundice until one-year-old.

Effect of Drug-containing Serum of Clerodendrum bungei on Liver Cancer Cells Through PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

Objective: To investigate the effect of Clerodendrum bungei-containing serum on liver cancer MHCC97-H cells and its possible mechanism from the perspective of phosphatidylinositol 3-kinase(PI3K)/protein kinase (Akt) signaling pathway. Method: The medicinal serum of 15% C. bungei was used to treat MHCC97-H cells. The effect of serum containing C. bungei on cell proliferation was observed by cell counting kit-8(CCK-8) method, in order to select the best time and concentration. The apoptosis was detected by Annexin V-FITC/PI double staining method. Western blot was used to detect the posphatase and tensin homologous gene deleted from chromosome 10 in key proteins (PTEN), phosphoprotein kinase B (p-Akt) and phosphatidylinositol 3-kinase (PI3K)-related protein expression of PI3K/Akt signaling pathway. Real-time PCR was used to detect C. bungei-containing serum on cells for 72 h after activation of nuclear factor-activated B cell kappa light chain(NF-κB) and tumor necrosis factor-α (TNF-α) mRNA expression. Result: The results of CCK-8 showed an inhibitory effect of the C. bungei-containing serum on the proliferation of tumor cells in a dose and time-dependent manner. Among them, the high-dose group had the most obvious inhibitory effect, and the maximum inhibition rates at 24, 48,72 h were 28%, 32%, and 43%, respectively. The results of flow cytometry showed that with the increase of drug-containing serum concentration, the cell growth was observed. The inhibition rate of cells was increased to different degrees, and the inhibition effect was significantly increased in the 72 h intervention group (PC. bungei-containing serum group was 19.48% and 19.72%, compared with the blank group. The difference was significant (PC. bungei-containing serum (PPC. bungei-containing serum could down-regulate the expression of NF-κB and up-regulate the expression of TNF-α mRNA (PConclusion: The medicinal serum of C. bungei can effectively inhibit the proliferation of MHCC97-H hepatoma cells and promote its apoptosis, which may be related to the PI3K/Akt signaling pathway and its key factors.

Significance of hepatitis B virus pregenomic RNA in the progression of chronic hepatitis B / 临床肝胆病杂志

Hepatitis B virus (HBV) pregenomic RNA (pgRNA) is the direct transcription product of HBV covalently closed circular DNA (cccDNA) and can reflect the transcriptional activity of HBV cccDNA and the progression of chronic hepatitis B, which provides guidance for clinical treatment and prognostic prediction. Compared with other common serological markers for HBV infection, HBV pgRNA is more sensitive in reflecting HBV replication and the effect of antiviral therapy has a certain predictive value for endpoints in the stages of antiviral therapy. This article elaborates on the significance of HBV pgRNA in reflecting the changes in disease conditions with reference to the correlation of HBV pgRNA with HBcrAg and HBV cccDNA.

Effects of Jianpi Xiaoai Prescription on Epithelial-mesenchymal Transition of Colorectal Cancer Cell SW620 Induced by TGF-β1 / 中国中医药信息杂志

Objective To observe the effects of Jianpi Xiaoai Prescription on epithelial-mesenchymal transition of colorectal cancer cell SW620 induced by TGF-β1; To discuss its possible mechanism of action for prevention and treatment of colorectal cancer. Methods Colorectal cancer cells SW620 were cultured in vitro. By conducting CCK-8 and Transwell experiments, the proliferation, invasion and migration of colorectal cancer cell (SW620) were detected. qRT-PCR and Western blot experiments were applied to verify the mRNA and protein expression level of E-cadherin and Vimentin. Results Jianpi Xiaoai Prescription showed in vitro inhibitory effect on the proliferation, invasion and migration of colorectal cancer cell SW620. Compared with blank group, the expression of E-cadherin in TGF-β1 induced group was reduced and Vimentin was increased (P<0.05); Meanwhile, the expression of E-cadherin in Jianpi Xiaoai Prescription group was increased and Vimentin was decreased when compared with TGF-β1 induced group (P<0.05). Conclusion Jianpi Xiaoai Prescription can inhibit the proliferation, invasion and migration of colorectal cancer cell by reverting the epithelial-mesenchymal transition of colorectal cancer cell induced by TGF-β1, with a purpose to achieve the goal of preventing and treating the recurring and migration of colorectal cancer.

Effect of Red Ginseng Extract on the Pharmacokinetics of Aspirin Metabolite in Sprague Dawley Rats / 中国医学科学杂志(英文版)

Objective To investigate whether red ginseng extract can affect the pharmacokinetics of aspirin in Sprague Dawley (SD) rats.Methods Totally, 12 male SD rats were randomly and uniformly divided into the aspirin group (aspirin 10.42 mg·kg ) and the combined group (red ginseng extraction 0.5 mg·g + aspirin 10.42 mg·kg ). After intragastric administration of drugs, blood samples (0.5 ml once) were drawn from orbit at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 10 and 12 hours after dosing. Plasma concentration of salicylic acid (metabolite of aspirin) was detected with ultraviolet-visible high performance liquid chromatography (HPLC). The reliability of the procedure was verified with respect to specificity, linearity, accuracy, precision, extraction recovery and matrix effect, and stability. Pharmacokinetics of salicylic acid was evaluated by using non-compartmental model. Results The method was proved to be validated. Compared with the aspirin group, area under the curve (AUC ) and maximum concentration of salicylic acid in rats of the combined group increased obviously (P<0.01), while clearance rate (CL/F) decreased clearly (P<0.05).Conclusion The in vivo study showed that red ginseng extract can help the internal absorption of aspirin, and delay the in vivo metabolism of aspirin.

Effect of pinostrobin chalcone on adipogenic differentiation of mesenchymal stem cell C3H10T1/2 / 中国中药杂志

Chalcones is a flavonoid wildly presented in many herbs. It has the effect to inhibit cells adipogenic differentiation. In order to study the effect of pinostrobin chalcone extracted and isolated from leaves of hickoryes on the adipogenic differentiation of murine embryonic mesenchymal stem cell (C3H10T1/2), MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)- 2H-tetrazolium] method was used to detect the cell proliferation; adipogenic differentiation was characterized by oil red O staining and isopropanol extraction; the triglyceride content was detected by GAP-PAP enzyme method; and the C3H10T1/2 cell differentiation into adipocytes was also examined by the mRNA and protein expression of PPARγ, C/EBPα and FABP4 by RT-PCR and Western blot respectively. Results indicated that pinostrobin chalcone almost had no effect on cell proliferation activity when the concentration was less than or equal to 50 μmol•L⁻¹; the oil red O staining, isopropanol extraction and GAP-PAP enzyme method showed that pinostrobin chalcone significantly decreased the C3H10T1/2 adipogenic differentiation and triglyceride content in the cytoplasm of adipocytes; the RT-PCR and Western blot analysis showed that pinostrobin chalcone can down-regulate the mRNA and protein levels of FABP4, PPARγ and C/EBPα in C3H10T1/2 cells(P<0.05 or P<0.01). The experiment results suggest that pinostrobin chalcone can inhibit C3H10T1/2 adipogenic differentiation.

Establishment of a mesenchymal stem cells model stably expressing hCD4 and hCCR5 / 重庆医学

Objective To establish a kind of mesenchymal stem cells(MSCs) model that could express hCD4 and hCCR5 to study the field of human immunodeficiency virus type 1 (HIV-1).Methods Lentiviral vectors containing the genes of hCD4 and hCCR5 under the transcriptional control of cytomegalovirus promoter were designed.MSCs were transfected by the lentiviral vectors at optimum multiplicity of infection.Transduction efficiencies of hCD4 and hCCR5 in MSCs were analyzed by quantitative real-time polymerase chain reaction(RT-PCR),Western blot,and immunofluorescence staining.Subsequently,the transfected human MSCs were infected with HIV-1,and the expression of HIVRNA in the MSCs was detected by RT-PCR.Results The MSCs model con-taining hCD4 and hCCR5 and supporting normal HIV-1 infection was constructed.qRT-PCR showed that MSCs upon infection with lentiviral vectors were highly expressed in hCD4 and CCR5 mRNA sequences(P＜0.01).Western blot detection showed the positive bands of 55.0 × 103 (hCD4) and 40.6 × 103 (hCCR5).The results of immunofluorescence staining revealed that hCD4 and CCR5 were expressed on the surface of MSCs.Such results were not found in cells infected with empty lentiviral vectors.And the susceptibility of the hCD4/CCR5 transgenic MSCs to the HIV-1 was further indicated by the detection of HIV-1 RNA in the culture supernatants and cell lysates(P＜0.05).Conclusion The MSCs model that could highly express hCD4 and hCCR5 was established to support normal HIV-1 infection,which can be used to investigate the development of new therapies and vaccines against HIV.

Multicenter clinical study of the predictive value of neutrophil gelatinase-associated lipocalin for contrast-induced acute kidney injury after emergency percutaneous coronary intervention in patients with relative normal renal function / 实用医学杂志

Objective To explore the predictive value of neutrophil gelatinase associated lipocalin (NGAL) for contrast-induced acute kidney injury (CI-AKI) after emergency percutaneous coronary interventions (PCI) in patients with relative normal renal function.Methods A total of 73 patients with relative normal renal function undergoing PCI were enrolled in this prospective multicenter clinical study.Serum NGAL was measured by point-of-care test.Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of NGAL for CI-AKI.Results CI-AKI occurred in 5 patients (6.85％).The concentration of NGAL was higher in the CI-AKI group than in the non-CI-AKI group.ROC curve indicated that the area under the receiver operating characteristic was 0.755,0.761 and 0.809,respectively.Conclusions Serum NGAL at baseline,4 h and 8 h after procedure are served as a good biomarker for early diagnosis of CI-AKI after PCI.Therefore,NGAL might become an early and quick marker for CI-AKI in the future.

Study on the rat pulmonary i rritation of aerosol inhaled Tanreqing and Reduning / 药学实践杂志

Objective To study the rat pulmonary irritant of aerosol inhaled Tanreqing and Reduning injection .Methods Rats were devided into two groups for each medicine (low concentration group and high concentration group ) ,nebulized drug administration for seven days ,with the control group irrigated with saline ,and were sacrificed .Through bronchoalveolar lav-age ,excurrent bronchoalveolar lavage fluid (BALF) was used for total protein determination and LDH vitality test to evaluate pulmonary toxicity of two medicines .Results The protein concentrations of two groups in low and high concentrations of Tan-reqing and Reduning respectively are (193 .78 ± 27 .74) ,(235.33 ± 50.41)μg/ml;(174 .02 ± 17 .82) ,(227 .27 ± 66 .03)μg/ml;LDH vitalities respectively are 1065 .21 ± 181 .76 ,1467 .33 ± 101 .87;307 .97 ± 47 .56 ,1377 .29 ± 566 .48 .By t-test ,compared with normal saline ,there was no significant effect among these five groups on protein concentration ,but these two medicine were able to improve LDH activity (P<0 .05) which was more obvious in high concentration group .When two medicines were in low concentration ,LDH activity was higher in Tanreqing group with statistical significance (P<0 .05) .Conclusion Aero-sol inhaled Tanreqing and Reduning injection in rats have some pulmonary irritation and potential safety hazard in this delivery w ay .

Prognostic values of common definition of contrast-induced nephropathy after coronary catheterization in patients with normal serum creatinine / 实用医学杂志

Objective To evaluate the prognostic values of common definition compared to traditional definition of contrast-induced nephropathy (CIN) in patients with normal serum creatinine (SCr). Methods Patients undergoing percutaneous coronary angiology or intervention with normal baseline SCr were enrolled prospectively. Those who were diagnosed as CIN according to common definition were divided into two groups based on the peak increase from baseline in the SCr concentration within 48 ~ 72 hours after the procedure: ≥ 44.2 μmol/L (CIN44.2 group, in common with traditional definition), ≥25% of baseline to < 44.2 μmol/L (CIN25%-44.2 group, interval between the two definitions). Hospital stay and long-term outcomes were compared among CIN44.2, CIN25%-44.2, and non-CIN groups. Results Of all 3,044 patients enrolled, 302 (9.9%) patients developed CIN according to common definition including CIN44.2 occurred in 56 (1.8%) patients and CIN25%-44.2 in 246 (8.1%) patients. Patients in CIN44.2 group indicated significant longer hospital stay and long-term outcomes compared with non-CIN group (P < 0.05). However, patients in CIN25%-44.2 group had similar in-hospital mortality and long-term cumulative risk of major clinical adverse events (MACE) and death with non-CIN group (all, P = 1.00). Multivariate Cox proportional hazard analyses also demonstrated that CIN25%-44.2 did not associate with long-term MACE (HR 1.16, P = 0.645) and death (HR 0.98, P = 0.964) after adjusting for potential confounding factors. Conclusions For patients with normal baseline SCr, common definition based on traditional definition of CIN is unreasonable and overestimates the incidence of CIN, whose extension of traditional denifition proves no significant clinical value.