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Objective @#To investigate the role of knockdown of peroxiredoxin-6 ( PRDX6) in injury and adaptive expression of bile acid transporter in human hepatoellular carcinomas ( HepG2 ) cells induced by rifampicin (RFP) . @*Methods @#Cells in logarithmic growth phase were uniformly inoculated in six-well plates , and HepG2 cells were transiently transfected with specific PRDX6-siRNA and control-siRNA to construct the knockdown group and control group . After 24 h of induction with 100 μmol/L RFP , Western blot and qRT-PCR were performed to detect the protein and gene expression levels of PRDX6 , multidrug resistance protein 1 (MDR1) , multidrug resist- ance-associated proteins 2 , 3 and 4 (MRP2 , MRP3 and MRP4) , and Na + /taurine taurocholate cotransporter pro- tein (NTCP) . Annexin V-FITC/PI double staining assay was used to detect the apoptosis rate of cells in each group ; CCK-8 assay was used to detect the changes of cell proliferation in each group; The relative contents of ala- nine aminotransferase ( ALT) , aspartate aminotransferase ( AST) , total bilirubin ( TBIL) , indirect bilirubin (IBIL) and total bile acid (TBA) in the supernatant of cell culture medium of each group were detected by kits .@*Results @#RFP increased the protein and gene expression levels of MRP2 , MRP3 , MRP4 , MDR1 , NTCP and PRDX6 in HepG2 cells (P < 0. 05) , while the protein and gene expression levels of MRP2 , MRP3 , MRP4 , MDR1 and NTCP decreased to different degrees after PRDX6 knockdown (P < 0. 05) . In addition , PRDX6 knockdown re- sulted in increased apoptosis rate of HepG2 cells (P < 0. 05) , decreased cell proliferation ability (P < 0. 05) , and increased levels of cell injury markers (ALT , AST , TBIL , DBIL , TBA) in cell culture supernatants (P < 0. 05) . @*Conclusion @#RFP increased the protein and gene expression of bile acid transporter and PRDX6 to increase in HepG2 cells . However , following knockdown of PRDX6 and treatment with RFP , the protein and gene expression levels of the bile acid transporter decreased and cell injury was aggravated , suggesting that PRDX6 played a protec- tive role in RFP-induced adaptive response in HepG2 cells .
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Objective @#To investigate the effect of mesencephalic astrocyte-derived neurotrophic factor (MANF) on the adaptive expression of bile acid transporter in human hepatoellular carcinomas (HepG2) induced by rifampicin (RFP) ．@*Methods@#The control group cell line (Y07) and the knockdown group cell line (Y25) were constructed by lentiviral stable transfection technology．The Y07 and Y25 cells were treated with RFP of 200 μmol / L for 48 h， and qRT-PCR and Western blot were used to detect the protein and gene expression levels of MANF，bile salt export pump ( BSEP) ，multidrug resistance-related proteins 2 /3 /4 ( MRP2 ，MRP3 ，MRP4) ，multidrug resistance protein 1 (MDR1) ，organic solute transporter a / β ( OSTα/ β) ，organic anion transporter ( OATP2B1) ．The protein and gene expression levels of proliferating cell nuclear antigen ( PCNA) ，proliferating cell marker Ki67 were used to evaluate the proliferation of cells in each group changes in levels．Changes in the protein and gene expression levels of C / EBP homologous protein( CHOP) and cysteinyl aspartate specific proteinase-3 ( Caspase-3) were used to evaluate the apoptosis of cells in each group．The relative contents of alanine aminotransferase(ALT) ，aspartate aminotransferase(AST) ，alkaline phosphatase ( ALP) ，total bilirubin ( TBIL) ，indirect bilirubin ( IBIL) and total bile acid(TBA) in the supernatant of cell culture medium of each group were detected by kits. @*Results@#RFP could induce the protein and gene expression of MANF，BSEP ，MRP2 ，MRP3 ，MRP4 ，MDR1 ，OSTα , OSTβ , OATP2B1 in HepG2 cells (P ＜0. 05 ) ，while the protein and gene expression levels of BSEP ，MRP2 ，MRP3， MRP4，MDR1，OSTα、OSTβ、OATP2B1 decreased after MANF knockdown(P＜0. 05) ．Moreover，under the action of RFP，the protein expression of PCNA and Ki67 in the knockdown group was still higher．The protein and gene levels of CHOP and Caspase-3 significantly increased after MANF knockdown(P＜0. 05) ．The levels of the hepatic cell injury markers in the cell supernatant increased significantly(P＜0. 05) ．@*Conclusion @#RFP can induce the expression of bile acid transporter such as BSEP，MRP2，MRP3，MRP4，MDR1，OSTα , OSTβ and OATP2B1 to increase in HepG2 cells(P＜0. 05) ，but the expression of bile acid transporter of HepG2 after MANF knockdown will significantly decrease under the induction of rifampicin(P＜0. 05) ，and cell indury is aggravated，indicating that MANF plays a protective role in RFP-induced adaptive responses by regulating the bile acid transporter.
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Pediatric cough is a heterogeneous condition in terms of symptoms and the underlying disease mechanisms. Symptom phenotypes hold complicated interactions between each other to form an intricate network structure. This study aims to investigate whether the network structure of pediatric cough symptoms is associated with the prognosis and outcome of patients. A total of 384 cases were derived from the electronic medical records of a highly experienced traditional Chinese medicine (TCM) physician. The data were divided into two groups according to the therapeutic effect, namely, an invalid group (group A with 40 cases of poor efficacy) and a valid group (group B with 344 cases of good efficacy). Several well-established analysis methods, namely, statistical test, correlation analysis, and complex network analysis, were used to analyze the data. This study reports that symptom networks of patients with pediatric cough are related to the effectiveness of treatment: a dense network of symptoms is associated with great difficulty in treatment. Interventions with the most different symptoms in the symptom network may have improved therapeutic effects.
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This study was aimed to analyze the treatment of pediatric cough disease by Professor Ni Zhuying and to provide clinical guidance for the treatment of pediatric cough disease.A total of 1640 cases of pediatric cough disease treated by Professor Ni Zhuying were selected.All cases were recorded into the structured clinical case acquisition system.Data mining platform was used in the data analysis.And the frequency statistics were used.Complex network analysis system was used to study the core prescription and traditional Chinese medicine (TCM) compatibility rules.The comparison of efficiency and the chi-squared test were used to analyze the core drug and the common compatibility.The results showed that when Professor Ni treated pediatric cough disease,the top 12 drugs used herbs were tangerine peel,fried bitter almonds,honey ephedra,fructus trichosanthis peel,radix peucedani,bran fried fructus aurantii,prepared rhizoma pinclliae,radix angelicae dahuricae,scutellaria,houttuynia,ramulus uncariae cum uncis,bran fried bombyx batryticatus.The core prescription results from the complex network system analysis was honey ephedra,fried bitter almonds,bran fried fructus aurantii,radix peucedani,fructus trichosanthis peel,dried tangerine peel,prepared rhizoma pinelliae,radix angelicae dahuricae.The commonly used drugs for compatibility were semen lepidii,rhizoma chuanxiong,berba asari;turmeric,raw rhubarb,periostracum cicadae,cortex lycii,vinegar prepared pericarpium citri reticulatae viride,cortex mori;radix astragali,bran fried rhizoma atractylodis macrocephalae,poria;bran fried medicated leaven,fried hawthorn;bulbusfritillariae thunbergii,radix scrophulariae;earthworm,ramulus euonymi,and etc.The drug efficacy and chi-squared test results showed that the main compatibility of drugs had more effect on the trend of efficacy.It was concluded that professor Ni's treatment of pediatric cough disease was mainly from the sputum,heat,and qi.Attention was paid to individual differences of children with syndrome differentiation.It can provide more reference to the treatment of clinical pediatric cough disease.
