Objective: To investigate the mechanism of curative effects of Guben Fangchuan capsule (固本防喘胶囊) on chronic obstructive pulmonary disease(COPD). Methods: Using double blind control method , 86 elderly COPD patients in in acute exacerbating stage were divided into treatment group and control group (n=43 in each group). The conventional treatment of the two groups was the same. Additionally, the patients in the treatment group were given Guben Fangchuan capsules orally. The levels of interleukinCD*28 (ILCD*28) and eotaxin in phlegm were detected by enzyme linked immunoadsorbent assay (ELISA). Chymase activity in phlegm was determined by spectrophotometry. They were compared before and after treatment. Results: ①The activity of chymase, the levels of ILCD*28, eotaxin, neutrophil (NEU) and eosinophil (EOS) in phlegm in patients with intermediate and severe COPD patients were lower after treatment in treatment group than those of the patients before treatment and control group (all P

Objective　To study the occurence, development and regulation of reactive gliosis with astrocyte (Ast) in vitro. Methods　Ast was isolated and cultured in vitro and its model of reactive gliosis was established by scratching the cultured astrocytes. The reactivity and rules of Ast to injury was studied by morphological changes, RT-PCR, immunocytochemistry, in situ hybridization and imaging analysis. Results　After scratching, the astrocytes showed typical features of reactive gliosis, with the hypertrophic cell body, thickened and lengtheded processes, and enhanced glial fibrillary acidic protein (GFAP) staining. In situ hybridization and RT-PCR analysis confirmed that the expression of GFAP mRNA was markedly increased. These changes occurred 1 d after scratching and reached the peak 5 to 7 d after injuring. Conclusion　A model of reactive astrogliosis was successfully established in vitro which showed an active reaction to injury. The characteristics of reactive gliosis parallel that seen in vivo.

Objective　To explore the role of tyrosine hydroxy lase (TH) gene in the treatment of Parkinson's disease (PD) by using ex vivo gene transfer. Methods　After the construction of TH gene in a retroviral vector, the astrocytes were cultured with the supernatant containing the recombinant DNA and then grafted into the cerebrum of PD rats. The reduction of the rat rotation beharior was evaluated. Results　The rotati on of PD rats was markedly improved in the rats with ex vivo gene transfer. Conclusion　TH has an obvious efficiency on the treatment of PD and the astrocytes can be used as effective gene transfer cells.

BACKGROUNDTo determine the inhibitory effect of antisense peptide nucleic acids (PNA) of telomerase on the growth of lung cancer cell lines.

METHODSThe synthesized modified antisense PNAs of telomerase were transfected into the lung cancer cell lines A549 and NCI-H446 respectively by lipofectamine transfection. Telomerase activity was detected by RT-PCR-ELLISA, and the cell counts were determined by MTT.

RESULTSSeventy two hours after transfection with antisense PNAs of telomerase, telomerase activity (A450 value) of A549 and NCI-H446 were down regulated from 0.582 ±0.039, 0.571±0.043 to 0.294±0.048 ( P < 0.01), 0.276±0.051 ( P < 0.01) respectively, and alive cell counts (A580 value) of them from 0.485± 0.009 , 0.513±0.015 to 0.191±0.027 ( P < 0.01), 0.138±0.046 ( P < 0.01) respectively. The growth of two lung cancer cell lines were significantly inhibited.

CONCLUSIONSAntisense PNA of telomerase might inhibit not only the telomerase activity, but also the growth of lung cancer cell lines in vitro.

OBJECTIVETo find out a possible approach to improve the effectiveness of radiotherapy and chemotherapy for Ewing's sarcoma by constructing a eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia and to evaluate the effects of this HRE regulated HSV-TK system on killing effect of gancyclovir (GCV) on Ewing's sarcoma cell line SK-ES under hypoxic condition.

METHODSThe HRE was synthesized according to the literature and cloned into the enhancer site of pIRES(2)-EGFP vector to obtain the pHRE recombinant plasmid. The HSV-TK was amplified by PCR and cloned into the multiple clone site of pIRES(2)-EGFP and pHRE to obtain pTK and pHRE-TK recombinant plasmid. The human Ewing's sarcoma cell line SK-ES was transfected by pTK or pHRE-TK recombinant plasmid with liposome and then was exposed to normoxic (21% oxygen) or hypoxic (3% oxygen) condition. The expression of enhanced green fluorescent protein (EGFP) was monitored by fluorescent microscopy. The sensitivity of human Ewing's sarcoma cell line SK-ES transfected with pTK or pHRE-TK recombinant plasmid to the anti-tumour drug GCV was determined with the method of tetrazolium (MTT) after treating with GCV for five days.

RESULTS(1) The result of sequencing showed that the recombinant plasmid pHRE contained HRE, and that the recombinant plasmid pTK and pHRE-TK contained HSV-TK gene in the sense direction. (2) Comparison of fluorescent optical density (FOD) showed that (1) the EGFP FOD value of pHRE and pHRE-TK group cells exposed to hypoxia was significantly higher than those exposed to normoxia (P < 0.01); (2) when the cells were exposed to hypoxia, the EGFP FOD value of pHRE and pHRE-TK group cells was significantly higher than that of pTK and empty vector group (P < 0.01); (3) there was no significant difference among the four groups of cells when they were exposed to normoxia (P > 0.05). (3) Comparison of the sensitivity of four groups of cells to GCV showed that (1) the cells in pHRE-TK and pTK groups were much more sensitive to GCV than the cells in pHRE group under hypoxia condition (P < 0.01), the higher the GCV concentration, the greater the difference; (2) the cells of pHRE-TK group were more sensitive to GCV than those in pTK group under hypoxic condition (P < 0.01), but was almost equally sensitive under normoxic condition (P > 0.05); (3) the pHRE-TK group cells had higher sensitivity to GCV under hypoxia than normoxia (P < 0.01) while the pTK group cells had almost the same sensitivity to GCV under hypoxia and normoxia (P > 0.05).

CONCLUSION(1) The eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia was constructed successfully. (2) HRE could up-regulate expression of EGFP by SK-ES cells under hypoxia condition. (3) HRE could enhance the killing effect of HSV-TK/GCV system on human Ewing's sarcoma cell line SK-ES under hypoxic condition.

Antiviral Agents ; pharmacology ; Cell Hypoxia ; drug effects ; genetics ; Cell Line, Tumor ; Ganciclovir ; pharmacology ; Gene Expression Regulation ; drug effects ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Microscopy, Fluorescence ; Plasmids ; Polymerase Chain Reaction ; Response Elements ; genetics ; Sarcoma, Ewing ; drug therapy ; metabolism ; Simplexvirus ; genetics ; metabolism ; Thymidine Kinase ; genetics ; metabolism ; Transfection

BACKGROUNDTo investigate the expression of the SOCS3 gene and its effect on proliferation of A549 cells.

METHODSA549 cells were cotransfected with pEFSOCS3 and pSV2neo by liposome, then G418 was used to screen the positive cells. Expression of SOCS3 mRNA and protein was detected by RT-PCR and immunocytochemistry respectively before and after transfection. MTT assay was used to detect the cell growth. Flow cytometric DNA analysis was used to determine the cell cycle.

RESULTSRT-PCR and immunocytochemistry showed that no expression of SOCS3 mRNA and protein was detected in A549 cells before transfection, but a stable expression of SOCS3 gene was observed after transfection with SOCS3 gene. Compared with control group, growth of A549 cells transfected with SOCS3 gene was significantly suppressed, with a suppressive rate of 41.07%. The cells at G₀/G₁ cell phases increased, and those at S and G₂/M phases decreased significantly after transfection.

CONCLUSIONSSOCS3 protein might inhibit the proliferation of A549 cells by negatively regulating cellular signal pathways.

OBJECTIVE: To investigate effects of berberine (BBR) on cholesterol synthesis in HepG2 cells with free fatty acid (FFA)-induced steatosis and to explore the underlying mechanisms. METHODS: A steatosis cell model was induced in HepG2 cell line fed with FFA (0.5 mmol/L, oleic acid:palmitic acid = 2:1), and then treated with three concentrations of BBR; cell viability was assessed with cell counting kit-8 assays. Lipid accumulation in cells was observed through oil red O staining and total cholesterol (TC) content was detected by TC assay. The effects of BBR on cholesterol synthesis mediators were assessed by Western blotting and quantitative polymerase chain reaction. In addition, both silent information regulator 1 (SIRT1) and forkhead box transcription factor O1 (FoxO1) inhibitors were employed for validation. RESULTS: FFA-induced steatosis was successfully established in HepG2 cells. Lipid accumulation and TC content in BBR groups were significantly lower (P < 0.05, P < 0.01), associated with significantly higher mRNA and protein levels of SIRT1(P < 0.05, P < 0.01), significantly lower sterol regulatory element-binding protein 2 (SREBP2) and 3-hydroxy 3-methylglutaryl-CoA reductase levels (P < 0.05, P < 0.01), as well as higher Acetyl-FoxO1 protein level (P < 0.05, P < 0.01) compared to the FFA only group. Both SIRT1 inhibitor SIRT1-IN-1 and FoxO1 inhibitor AS1842856 blocked the BBR-mediated therapeutic effects. Immunofluorescence showed that the increased SIRT1 expression increased FoxO1 deacetylation, and promoted its nuclear translocation. CONCLUSION BBR can mitigate FFA-induced steatosis in HepG2 cells by activating SIRT1-FoxO1-SREBP2 signal pathway. BBR may emerge as a potential drug candidate for treating nonalcoholic hepatic steatosis.
Berberine/pharmacology* ; Cholesterol ; Forkhead Box Protein O1/genetics* ; Humans ; Non-alcoholic Fatty Liver Disease/drug therapy* ; Sirtuin 1/genetics* ; Sterol Regulatory Element Binding Proteins

Anaplastic lymphoma kinase (ALK) fusion gene, as a tumor driver gene, was crucial for the occurrence and development of non-small cell lung cancer (NSCLC). Recently, targeted ALK fusion gene has become the main treatment method for ALK-positive NSCLC. The first and second generation ALK inhibitors (ALKi), such as crizotinib, ceritinib, alectinib and ensartinib have been approved in China. However, there was no guidance for the management of ALKi adverse reactions. Therefore, this "Recommendations from experts in the management of adverse reactions to ALK inhibitors (2021 version)" has been summarized, led by Lung Cancer Professional Committee of Sichuan Cancer Society and Sichuan Medical Quality Control Center for Tumor Diseases, to provide practical and feasible strategies for clinical ALKi management specification of adverse reactions.
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Carcinoma, Non-Small-Cell Lung ; Crizotinib ; Humans ; Lung Neoplasms/genetics* ; Protein Kinase Inhibitors/adverse effects* ; Receptor Protein-Tyrosine Kinases/antagonists & inhibitors*

Aortic Valve/surgery* ; Aortic Valve Stenosis/surgery* ; China/epidemiology* ; Heart Valve Prosthesis Implantation ; Humans ; Severity of Illness Index ; Treatment Outcome