Objective To study any change in the expression of γ-glutamylcysteine synthetase (γ-GCS) in the skeletal muscles of mice with emphysema.MethodsTwenty Kunming male mice were divided into a normal control group and an emphysema group (n =10 in each ).An emphysema model was established by passive cigarette smoking in 8 of the emphysema group mice.TUNEL staining was used to detect apoptotic skeletal muscle cells.Im munohistochemical assays were used to determine the protein level of γ-GCS synthetase.ResultsThe average optical density of apoptotic cells was significantly higher in the skeletal muscles of the mice with emphysema compared with the normal controls,and their average γ-GCS synthetase levels were also significantly higher.Conclusions Apoptotic cells increase in skeletal muscle during emphysema,which may be caused by an oxidation/antioxidant imbalance mediated by γ-GCS synthetase.

Objective To study the changes in mRNA and protein expression of ubiquitin and to explore the relationship of apoptosis in the skeletal muscle of mouse with emphysema.Methods Emphysema model was established by passive cigarette smoking in mouse.Apoptosis was detected with TUNEL staining.The reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical assay were used to determine the mRNA and protein level of ubiquitin.Results Apoptotic cells were increased in skeletal muscle of mouse with emphysema.The mRNA and protein level of ubiquitin were significantly higher in mouses with emphysema (0.48±0.02 and 0.23+0.05,respectively) than in control group (0.17±0.01 and 0.14+0.01,t=6.223、4.093,both P ＜0.05).Conclusions The increase of apoptosis in skeletal muscle of mouses with emphysema may be associated with high expression of ubiquitin.

Objective To study the correlation between structure of b D -glucan derivatives from Poria cocos sclerotiuma and anti gastric cancer activities of elderly rats.Methods A water insoluble (1→3)-β-D-glucan (PCS3-Ⅱ ) isolated from fresh sclerotium of Poria cocos was sulfated,carboxymethylated,methylated,hydroxyethylated and hydroxypropylated,respectively,to prepare five water-soluble derivatives.Their activities of the native β-glucan and five derivatives against gastric cancer cell strains of MKN-45,SGC 7901and MKN-28 were tested in vitro and in vivo.Results The native β-glucan did not show any anti-gastric cancer activity,while the sulfated and carboxymethylated derivatives significantly exhibited the anti gastric cancer activity against MKN-45,SGC-7901 and MKN-28 cells in vitro,and elderly rats with MKN-45 transplanted tumor,especially.The gastric cancer inhibition rates of elderly rats were 32.7％ and 36.4％ for the 100 mg/kg sulfated and carboxymethylated derivatives,respectively.Conclusions The fresh sclerotium of Poria cocos polysaccharides can not show any anti gastric cancer activity in vitro,but the sulfated and carboxymethylated derivatives may increase the inhibition effect.Water-solubility,higher chain stiffness and relatively molecular weight would be benefit to increase anti-elderly gastric cancer activity of polysaccharides from Poria cocos sclerotiuma.

Objective To investigate the expression of anoctamin 1 in Chinese hamster ovary cells (CHO)and to analyze the functional properties of its ion channel,and to provide experimental basis for study on the physiological function of calcium-activated chloride channel.Methods The whole sequence of anoctamin 1 was obtained by PCR technique and was subcloned into pcDNA3.1 to construct the expression vector pcDNA3.1-anoctamin 1 was transfected into CHO by liposome-mediated transfection and the CHO stably expressing anoctamin 1 were aquired by selection with zeocin. The expression and distribution of anoctamin 1 in CHO were measured by RT-PCR technique and inverted fluorescence microscope.The functional properties of anoctamin 1 were measured with halide-sensitive fluorescence proteins YFP-H148Q/I152L.The PBS buffer solution with calcimycin and high concentration of iodine ion was used as experimental group,andthe PBS buffer solution without calcimycin and high concentration of iodine ion was used as control group.Results The results of digestion and sequencing confirmed that anoctamin 1 was cloned into pcDNA3.1 by electrophoresis and blast. The results of RT-PCR and inverted fluorescence microscope indicated that anoctamin 1 was expressed in CHO. The results of I- influx as measured by halide-sensitive fluorescence proteins YFP-H148Q/I152L showed that anoctamin 1 had the more functional properties of trans-epithelial transporting I-,and the transporting speed in experimental group was higher than that in control group (P<0.05).Conclusion Anoctamin 1 can be expressed highly in the CHO;Anoctamin 1 expressed in CHO has the characteristics of calcium-activated chloride channel.

Objective To study the effect of Angong-niuhuang pill on expression of eNOSmRNA in spontaneously hypertensive rats after intracerebral hemorrhage and to explore it's protective mechanism.Methods 120 spontaneously hypertensive rats were randomly divided into four groups: model group, western medicine treated group. Angong-niuhuang pill treated group, combining traditional Chinese medicine western medicine group, expression of eNOSmRNA in rat brain tissue was detected by RT-PCR. Results Expression of BCL-2mRNA in combining traditional Chinese medicine western medicine group was significantly higer than other goups(P＜0.01), Compared with model group, the expression of BCL-2mRNA in western medicine treated group and Angong-niuhuang pill treated group was markedly increased (P＜0.01), the differences between Angong-niuhuang pill treated group and Western medicine treated group were not obvious (P＞0.05).Conclusion Protective effect of Angong-niuhuang pill on spontaneously hypertensive rats after intracerebral hemorrhage is related to the the inducement of eNOS expression.

Objective: Predefining the most effective treatment for patients with depressive disorders remains a problem. We will examine the differential brain regions of gray matter (GM) in major depressive disorder (MDD) patients and the relationship between changes in their volume and the efficacy of early antidepressant treatment using magnetic resonance imaging (MRI). Methods: 159 never-medicated patients with first-episode MDD and 53 normal control subjects (NCs) were enrolled. The brains were scanned by MRI and measured with the 17-item Hamilton Depression Rating Scale (HAMD-17) at baseline and after 2 weeks of treatment with selective serotonin reuptake inhibitor (SSRI)s, and the non-responder group and responder group were obtained. The patients were analyzed by voxel-based morphological (VBM) and SPSS software. Receiver operator characteristics (ROC) analysis was performed for the difference between the responder group and the non-responder group in the differential brain regions, and Pearson correlations were computed between volume size and HAMD score reduction rate. Results: Smaller GM volume of the right superior temporal gyrus (STG), and the orbital parts of the right medial frontal gyrus and right inferior frontal gyrus were observed in MDD versus the NCs. The non-responder group demonstrated a significant volume reduction at the right STG compared with the responders, but no corresponding change in orbital part of right medial frontal gyrus and right inferior frontal gyrus. ROC analysis showed that Accuracy=71.2%. There was a positive correlation between the STG gray matter volume and the HAMD-17 score reduction rate (r=0.347, p=0.002). Conclusion The study results confirmed the local changes in brain structure in MDD and may initially predict the early treatment response produced by SSRIs as antidepressants.

ObjectiveTo provide a new idea for exploring the molecular genetic approach to the pathogenesis of schizophrenia via construction of microRNA-messenger RNA （miRNA-mRNA） regulatory network in schizophrenia. MethodsThe microarray datasets of GSE54578 miRNA expression profiles in peripheral blood and GSE145554 mRNA expression in the anterior cingulate in postmortem brain of schizophrenic subjects were downloaded from Gene Expression Omnibus （GEO） database since July 2021. The GEO2R was used to identify the differentially expressed miRNAs and mRNAs， screen the miRNA with target differentially expressed mRNA， and predict their potential upstream transcription factors. The overlapping genes from the mRNA targeted by the differentially expressed miRNA and the mRNA differentially expressed in GSE145554 dataset were collected. Then the biological features of hub genes were analyzed via Gene Ontology （GO） analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis， and the protein-protein interaction （PPI） network and miRNA-mRNA regulatory network of hub genes were constructed. ResultsA total of 8 up-regulated differentially expressed miRNAs with targeted mRNA were screened out in GSE54578 datasets regarding schizophrenia， which involved in the regulation of 10 transcription factors， 247 down-regulated differentially expressed mRNAs were screened out in GSE145554 datasets， and 17 overlapping mRNAs were obtained. GO analysis showed that the target mRNAs were mainly involved in astrocyte differentiation and development. KEGG pathway enrichment analysis showed that the target mRNAs were mainly involved in Rap1 and Ras signaling pathways. PPI network analysis showed that the mRNAs （KRAS and CD28） might be key genes in schizophrenia. ConclusionThe integrated bioinformatics analysis based on GEO database can identify potential susceptibility genes in schizophrenia， and it also contributes to the construction of miRNA-mRNA regulatory network in schizophrenia.

Objective: To understand the epidemiologic features of the rabies in Xishuang banna prefecture of Yunnan province, China in 2008-2017 and the viral molecular-evolution characteristics. Methods: The data of rabies case questionnaire were collected. The brain tissue samples from mad dogs, suspicious sick dogs and human brain tissue, saliva and cerebrospinal fluid samples from rabies patients were collected in Xishuangbanna. Coding region of nucleoprotein and glycoprotein genes were amplified by RT-PCR and sequenced. Homology and phylogenetic analysis were performed using the relevant bioinformatics software. Results: A total of 62 cases of human rabies were occurred in 28 districts of the 3 counties, Xishuangbanna prefecture in 2008-2017. Of them, 37 cases in Jinghong county, 15 in Menghai county and 10 in Mengla county. In which 48 cases were bitten by domestic dogs (77.42%), 11 cases were bitten by wild dogs (17.74%). Rabies case was occurred every year in the past decade. The seasonal incidence was not obvious. The majority of patients were aged from 30 to 59 years-old, with the youngest 1 year-old and the eldest 91 year-old. The male to female ratio was 1.70∶1, most cases were farmers. The nucleotide sequences of nucleoprotein gene of 9 virus strains (7 from Jinghong, 1 from Menghai and 1 from Mengla) were obtained from the samples of dogs and patients. Homology and phylogenetic analyses indicated that the 5 strains belonged to clade China-Ⅰ, 3 clade China-Ⅱ and 1 clade China-Ⅵ. The nucleotide sequences of glycoprotein gene of 5 virus strains (3 from Jinghong, 1 from Menghai and 1 from Mengla) were obtained from these positive samples, and all were clade China-Ⅰ, it is same with nucleoprotein genes analysis result from these 5 virus strains. These China-Ⅰ and China-Ⅱ strains from Xishuangbanna have a closer genetic relationship with same clade strains isolated from Pu’er and other prefectures of Yunnan province as well as Sichuan, Guizhou and Guangxi. The China-Ⅵ strain from Xishuangbanna share high homology and genetic relationship with China-Ⅵ strains isolated from southwestern Yunnan and neighbouring countries such as Myanmar, Laos and Vietnam in recent years. Conclusions In Xishuangbanna, rabies mainly occurred in rural area and domestic dog was the main source of transmission. These RABV clades China-Ⅰ, China-Ⅱ and China-Ⅵ were found in this region and the China-Ⅰ was principal clade. The transmission source of China-Ⅰ and China-Ⅱ were from adjacent areas in the province and China-Ⅵ was from Myanmar and Laos.