Objective To investigate the expression of aquaporin-5(AQP5) in lipo-polysaccharide(LPS)-cigarette smoking inducible SD rats,and the effect of ambroxol chloride(AMB)on its expression. Methods Twenty-one SD rats were randomly divided into three groups: AMB intervention group,model group(LPS-cigarette smoking induction group) and control group.TNF-? was determined from lung homogenate supernatant,bronchial alveolar lavage fluid(BALF) and serum by ELISA.The semi-quantitation of AQP5 transcription and expression were measured by in situ hybridization and immunohistochemistry,respectively. Results TNF-? from lung homogenate supernatant and BALF in model group was more than AMB intervention group and control group(P

OBJECTIVETo study the protective effect of Sanhuangyinchi Fang drug serum (SF) against hydrogen peroxide-mediated DNA oxidative damage in LO2 cells.

METHODSThe LO2 cells were randomly divided into the control group, H(2)O(2) group, SF groups (5%, 10%, and 15%) and vitE group. The morphological features of the treated LO2 cells were observed under inverted microscope. The viability of the treated cells was assessed with CCK-8 method, and the activity of SOD, CAT and GSH-PX were detected biochemically. Reactive oxygen species (ROS) levels, the content of 8-OHdG, and DNA damage of the cells were evaluated by flow cytometry, ELISA, and Comet assay, respectively.

RESULTSCompared with H(2)O(2) group, the cells in SF groups (10% and 15%) and vitE group showed higher cell survival rate (P<0.05) and higher SOD, CAT, GSH-PX (P<0.05) and ROS scavenging activities (P<0.01) with markedly decreases the content of 8-OHdG (P<0.01) and reduced tailing ratio, tail length, tail moment and Olive tail moment (P<0.05).

CONCLUSIONSF drug serum, especially at the concentration of 15%, can protect LO2 cells from H(2)O(2)-mediated DNA oxidative damage.

Cell Line ; Comet Assay ; DNA Damage ; Deoxyguanosine ; analogs & derivatives ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Hydrogen Peroxide ; toxicity ; Oxidation-Reduction ; Oxidative Stress ; Protective Agents ; pharmacology ; Reactive Oxygen Species

OBJECTIVETo study the effects of extracts of root of kudzu vine on mammary gland and uterus development in rats.

METHOD40 Wistar rats weighting 65-85 g were randomly divided into 4 groups: control group, estrogen group, extracts of root of kudzu vine group of high dose, extracts of root of kudzu vine group of low dose. (10 rats in each group). After having been treated for 7 days, the rats were killed; mammary glands and uterus were removed and weighed. Serum was isolated and kept at 4 degrees C for determination of hormones.

RESULT1. Administration of the root of kudzu vine significantly increased the weigh of mammary gland and uterus in rats. 2. Administration of the root of kudzu vine increased serum FSH, LH, E2 and decreased PRL.

CONCLUSIONExtracts of root of kudzu vine could enhance the weight of mammary gland and uterus growth in rats, which may provide experimental evidence for the development of new drug used for promoting mammary gland and uterus.

Animals ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Estradiol ; blood ; Female ; Follicle Stimulating Hormone ; blood ; Luteinizing Hormone ; blood ; Mammary Glands, Animal ; anatomy & histology ; growth & development ; Organ Size ; drug effects ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Prolactin ; blood ; Pueraria ; chemistry ; Rats ; Rats, Wistar ; Uterus ; anatomy & histology ; growth & development

The aim of study was to establish the packaging system of the recombinant lentiviral vector encoding Gfi1 gene for eukaryotic expression and to realize the efficient, stable expression of Gfi1 32D cells so as to provide effective platform for further studying the development of Gfi1 gene in hematologic malignancies. The three-plasmid recombinant lentiviral vector consisting of transfer plasmid (pLOX-Gfi1/pLOX), the packaging plasmid (pCMVDeltaR8.2) and the envelop plasmid (pMD.G) was prepared and purified. Human embryonic kidney 293T cells were cotransfected with the three plasmids by lipofectamine 2000. After transfection for 48 hours, the viral supernatant was collected and the target cell 32D was transfected with the recombinant lentivirus; the Gfi1 integration and expression in 293T and 32D cells were detected by Western-blot. The results showed that the three plasmids of lentivirus could be transfected into 293T with high efficiency and packaged successfully, and the Gfi1 protein could be detected by fluorescent microscopy. The recombinant lentiviruses carrying Gfi1 could transfer and deliver Gfi1 gene to 32D cells, and the Gfi1 expression in 293T and 32D cell could be detected by Western blot. It is concluded that the recombinant lentivirus carrying Gfi1 can deliver target gene to 32D cells with high efficiency, and the expression of Gfi1 protein is stable in 32D.
Cell Line ; DNA-Binding Proteins ; genetics ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; Transcription Factors ; genetics ; Transfection

OBJECTIVEIn order to explore the existence of SARS coronavirus (Co-V) and/or its RNA in sewage of hospitals administered SARS patients.

METHODSA novel electropositive filter was used to concentrate the SARS-CoV from the sewage of two hospitals administered SARS patients in Beijing, including twelve 2,500 ml sewage samples from the hospitals before disinfection, and ten 25,000 ml samples after disinfection; as well as cell culture, RT-PCR and sequencing of gene to detect and identify the viruses from sewage.

RESULTSThere was no live SARS-CoV detected in the sewage in this study. The nucleic acid of SARS-CoV had been found in the 12 sewage samples before disinfection from both hospitals by semi-nested PCR. After disinfection, SARS-CoV RNA could only be detected from the samples from the 309th Hospital, and the others were negative.

CONCLUSIONIt provides evidence that there is no live SARS-Cov in the sewage from hospitals with SARS patients though SARS-CoV RNA can be detected.

Hospitals ; Humans ; Nucleocapsid ; analysis ; RNA, Viral ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; Severe Acute Respiratory Syndrome ; virology ; Sewage ; virology

OBJECTIVE: To investigate the phenolic composition, antioxidant properties, and hepatoprotective mechanisms of polyphenols from green tea extract (GTP) in carbon tetrachloride (CCl)-induced acute liver injury mouse model. METHODS: High-performance liquid chromatography was used to analyze the chemical composition of the extract. Antioxidant activity of GTP was assessed by O, OH, DPPH, and ferric-reducing antioxidant power (FRAP) assay in vitro. Sixty Kunming mice were divided into 6 groups including control, model, low-, medium-, and high-doses GTP (200, 400, 800 mg/kg) and vitamin E (250 mg/kg) groups, 10 in each group. GTP and vitamin E were administered at a level of abovementioned doses twice per day for 7 days prior to exposure to a single injection of CCl. Hepatoprotective effects of GTP were evaluated in a CCl-induced mouse model of acute liver injury, using commercial enzyme linked immunosorbent assay kits, histopathological observation, terminal deoxynucleotidyl transferase-mediated dUTPNick-end labeling (TUNEL) assay and Western blot. RESULTS: GTP contained 98.56 µg gallic acid equivalents per milligram extract total polyphenols, including epicatechingallate, epigallocatechin gallate, epicatechin, and epigallocatechin. Compared with the model group, low-, medium-, or high doses GTP significantly decreased serum levels of alanine aminotransferase and aspartate transaminase (P<0.01). Histopathological observation confirmed that pretreatment of GTP prevented swelling and necrosis in CCl-exposed hepatocytes. Hepatoprotective effects of low-, medium-, and high-dose GTP were associated with eliminating free radicals and improving superoxide dismutase, catalase, and glutathione peroxidase activity in the liver. Additionally, low-, medium-, and high-dose GTP decreased cell apoptosis in the CCl-exposed liver (P<0.01). Phosphorylated nuclear factor kappa-B (NF-κB), p53, Bcl-2 associated x protein/B-cell lymphoma/leukemia-2 gene, cytochrome C, and cleaved caspase-3 levels were downregulated compared with the model group (P<0.01). CONCLUSION GTP achieves hepatoprotective effects by improving hepatic antioxidant status and preventing cell apoptosis through caspase-3-dependent signaling pathways.

Objective To discover critical genes contributing to the stemness and maintenance of spermatogonial stem cells (SSCs) and provide new insights into the function of the leucine-rich repeat (LRR) family member Lrrc34 (leucine-rich repeat-containing 34) in SSCs from mice. Methods Bioinformatic methods, including differentially expressed gene (DEG), gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, were used to uncover latent pluripotency-related genes. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence analyses were utilized to verify the mRNA and protein expression levels, respectively. RNA interference of Lrrc34 using siRNA was performed to detect its transient impact on SSCs. Results Eight DEGs between ID4-EGFP⁺ (G) and ID4-EGFP⁺/TSPAN8^High (TH), eight DEGs between G and ID4-EGFP⁺/TSPAN8^Low (TL) and eleven DEGs between TH and TL were discovered, and eleven protein-protein interaction (PPI) modules were found to be significant in the PPI network of DEGs. One of the DEGs, Lrrc34, was selected as a potential pluripotency-related gene due to its differential expression among ID4-EGFP⁺ spermatogonia subsets and its interaction with fibroblast growth factor 2 in the fifth module. Immunofluorescence experiments exhibited specific expression of Lrrc34 in a subpopulation of undifferentiated spermatogonia marked by LIN28A, and RT-PCR experiments confirmed the high expression of Lrrc34 in SSCs from P7 and adult mice. The transient knockdown of Lrrc34 in SSCs resulted in reduced colony sizes and significant changes in the transcriptome and apoptotic pathways. Conclusion Lrrc34 is highly expressed in mouse SSCs and is required for SSC proliferation in vitro through effects on transcriptome and signaling transduction pathways.
Animals ; Apoptosis/genetics* ; Cell Proliferation/genetics* ; Cells, Cultured ; Gene Expression Profiling/methods* ; Gene Ontology ; Gene Regulatory Networks ; Humans ; Male ; Mice, Inbred C57BL ; Mice, Transgenic ; RNA Interference ; Repressor Proteins/metabolism* ; Signal Transduction/genetics* ; Stem Cells/metabolism*