BACKGROUND:Ion channel analytical technique has verified that huwentoxin is an N-type Ca2+channel blocker affecting on presynaptic membrane. OBJECTIVE:To observe the effects of N-type Ca2+channel blocker huwentoxin on expressions of tumor necrosis factorα, tumor necrosis factor receptor I, tumor necrosis factor receptor-related death domain, Fas-related death domain protein and Caspase 8 in the hippocampi of rat models of global cerebral ischemia reperfusion injury. METHODS:Rat models of global cerebral ischemia and subarachnoid catheter were established using Pulsinel i 4-vessel occlusion and then received infusion of huwentoxin or normal saline via a PE10 tube. Morphological changes in the mitochondria and ultrastructure of pyramidal neurons in the hippocampal CA1 region of rats with global cerebral ischemia reperfusion injury were observed using electron microscope. The expressions of tumor necrosis factorα, tumor necrosis factor receptor I, tumor necrosis factor receptor-related death domain, Fas-related death domain protein and Caspase 8 were measured using RT-PCR. RESULTS AND CONCLUSION:Huwentoxin could maintain the basic morphology of mitochondria of rats with global cerebral ischemia reperfusion injury and decrease the expressions of tumor necrosis factorα, tumor necrosis factor receptor I, tumor necrosis factor receptor-related death domain, Fas-related death domain protein and Caspase 8 mRNA. Results suggested that huwentoxin as a novel N-type Ca2+channel blocker could block extracellular Ca2+influx, reduce intracellular Ca2+concentration, diminish a series of pathological lesion induced by intracellular Ca2+overload, protect nerve cells, and lessen the injury to nerve cells of hippocampus after ischemia and hypoxia.

OBJECTIVE:To study the hepatoprotective effect of the extract of Sabia parviflorawall(SPS) on mice with experimental liver injury.METHODS:The liver injury models were reproduced in mice with CCL4 and Paracetamol(AAP),respectively with the activities of serum ALT and AST measured.RESULTS:The activities of serum ALT and AST were all reduced to a certain degree in mice with liver injury induced by either CCL4 or Paracetamol(AAP).CONCLUSION: SPS exhibited certain hepatoprotective effect.

Objective To investigate specific anti-leukemia immune response induced by tumor lysates pulsed dendritic cells(TP-DC)in syngeneic bone marrow-transplanted mice.Methods From June 2002 to June 2003,the bone marrow cells were induced by using granulocyte-macrophage colony stimulating factor(mGM-CSF)and interleukin-4(mIL-4).DC was pulsed with L7212 tumor lysates at the immature stage.TP-DC was harvested on 7th day.615 mice immunized thrice with L7212 TP-DC starting at day 7 following BMT.Additional control groups were set up.The survival was observed.Results Immunization using TP-DC could induce L7212 cells-specific CTL responses that were statistically significant compared with control groups(F=391.77,P

Objective To investigate whether two polymorphism sites of the the exon 2 Lys65Lys(G/A) and exon 9 Va1365Met(G/A) in T ceils immunoglobulin domain and mucin domain protein-4(TIM-4) are associated with asthma in Chinese Han population of Hubei province. Methods The polymorphisms were de-tected with polymerase ehain reaction-restriction fragment length polymorphism(PCR-RFLP) in 185 cases of al-lergic asthma and 162 healthy controls. The genotype and allele frequencies were calculated and analyzed. Re-sults(1)The genotype frequencies of G/G, G/A and A/A in Lys65Lys(G/A)polymorphism were 0.840, 0.160 and 0 respectively in the healthy population, and were 0.859, 0.141 and 0 respectively in the allergic asthma population. No significant difference in genotype and allele frequencies was found between asthma pa-tients and the control subjects (P=0.603, P=0.618). (2) The polymorphism of the Val365 Met(G/A) was not detected in our study. Conclusion There is polymorphism site of the exon 2 Lys65Lys(G/A)in TIM-4, but this polymorphism site is not associated with asthma in Han nationality in Hubei Chinese population. There is no SNP of the exon 9 Val365Met(G/A) in TIM-4 in Chinese Han population of Hubei province.

Objective: To investigate the prevalence of hepatitis B virus (HBV) carriage among pregnant and lying-in women in Cangnan County, Zhejiang Province from 2011 to 2022 and identify the influencing factors, so as to provide insights into the guidance of healthcare among HBV carriers during pregnancy. Methods: A total of 34 403 women delivered in The Third People's Hospital of Cangnan County from January 2011 to July 2022 were enrolled, and their demographics, HBV carriage and pregnant outcomes were collected. The prevalence of HBV carriage was analyzed among pregnant and lying-in women, and factors affecting HBV carriage were identified using a multivariable logistic regression model. Results: A total of 34 403 pregnant and lying-in women were enrolled, with a median age of 27.00 (interquartile range, 7.00) years, and including 8 118 floating populations (23.60%). The overall prevalence of HBV carriage was 3.44%, and the prevalence of HBV carriage was 1.59% from 2011 to 2014, 4.08% from 2015 to 2018 and 6.86% from 2019 to 2022, appearing a tendency towards a rise (P<0.05). Multivariable logistic regression analysis identified estimated age of delivery (20-24 years, OR=1.832, 95%CI: 1.037-3.235; 25-29 years, OR=2.404, 95%CI: 1.372-4.214; 30-34 years, OR=2.914, 95%CI: 1.656-5.129; 35-39 years, OR=3.116, 95%CI: 1.741-5.576; 40 years and older, OR=2.358, 95%CI: 1.145-4.858), floating population (OR=0.670, 95%CI: 0.574-0.782), scarred uterus after cesarean section (OR=1.228, 95%CI: 1.076-1.521) and year of delivery (from 2015 to 2018, OR=2.504, 95%CI: 2.143-2.926; from 2019 to 2022, OR=4.425, 95%CI: 3.779-5.182) as factors affecting HBV carriage among pregnant and lying-in women. Conclusions The prevalence of HBV carriage rate appeared a tendency towards a rise among pregnant and lying-in women in Cangnan County from 2011 to 2022. Estimated age of delivery, floating population, year of delivery and scarred uterus after cesarean section are factors affecting HBV carriage.

RNA interference(RNAi), a highly conserved process of post-transcriptional gene silencing, can be induced by small interfering double-stranded RNA that mediate sequence-specific mRNA degradation. In the past several years, RNAi has been widely used as both an experimental tool to study mammalian gene function and a potential therapeutic approach to treat human diseases. In addition, some new proteins which involve in RNAi pathway have been characterized in mammalian cells. Here, we summarize the various molecules in RNAi, mechanism of action, and the current therapeutic applications in cancers.

PURPOSE: This study was conducted to investigate the small double-stranded RNA (dsRNA) mediated anti-tumor effects of p21(WAF1/ClP1) (p21) transcriptional activation in vitro in the human glioma SHG-44 cell line. MATERIALS AND METHODS: Human glioma SHG-44 cells were transfected with dsRNA using LipofectAMINE 2000 transfection reagent. Real-time PCR and Western blot analysis were conducted to detect p21 and survivin mRNA and protein levels, respectively. Cell proliferation was examined by MTT assay. Cell cycle distribution and apoptosis were detected by flow-cytometric analysis. RESULTS: We found that dsRNA targeting p21 promoter (dsP21) significantly induced the expression of p21 at transcription and protein levels, and reduced the expression of survivin. AS well, dsP21 transcription significantly inhibited human glioma SHG-44 cell proliferation. Analysis of cell cycle distribution revealed that dsP21 transfection increased accumulation of cells in the G0/G1 phase and reduced accumulation of cells in the S phase. Further analysis revealed that dsP21 transcription led to an increase in both early and late stages of apoptosis in human glioma SHG-44 cells. CONCLUSION: In the present study, P21 activation by RNA-induced gene activation (RNAa) induced anti-tumor activity in vitro in a human glioma SHG-44 cell line. The results suggested that RNAa could be used for human glioma treatment by targeted activation of tumor suppressor genes.
Apoptosis ; Blotting, Western ; Cell Cycle ; Cell Line* ; Cell Proliferation ; Genes, Tumor Suppressor ; Glioma* ; Humans* ; Methods ; Real-Time Polymerase Chain Reaction ; RNA, Double-Stranded* ; RNA, Messenger ; S Phase ; Transcriptional Activation* ; Transfection

Objective To observe the mRNA expressions of endothelin-1(ET-1) and endothelin A/B receptors (ETA/B) in tissue of benign prostatic hyperplasia treated by daily low-dose sildenafil.Methods A total of 32 patients with benign prostatic hyperplasia were randomly divided into two groups:treatment(25 mg sildenafi for 12 weeks,n=16) and control (no drug,n=16) groups.Immunohistochemical staining,ELISA and RT-PCR were used to detect the expression levels of ET-1 protein and ET A/B mRNA,respectively.Results The expressions of ET-1 protein and ET A/B mRNA in prostatic tissue were significantly lower in treatment group than in control group[(53.31±18.56) ng/kg vs.(83.34±31.38) ng/kg,0.356±0.056 vs.0.624±0.083,0.721±0.083 vs.0.933±0.905,t=-3.295,10.715,6.937,all P＜0.001].Conclusions Daily low-dose sildenafil can reduce the expressions of ET-1 and ET A/B receptors mRNA in benign prostatic hyperplasia.

ObjectiveTo optimize Chonglian oral solution extracting craft. MethodsWith the obtaining rate of extract and the total content of the Pariphyllin Ⅰ and Phillyrin presented in the extract as the indexes for the water extraction process,and with the total content of the Pariphyllin Ⅰ and Phillyrin presented in the extract as the indexes for the alcohol deposition process,orthogonal design was used to optimize the conditions for the extraction process respectively. ResultsThe optimal conditions for the water extraction of Chonglian oral solution was as following:to add water 10 times,decocting 3 times,1 hour each time.The optimal conditions for the alcohol deposition of Chonglian oral solution was as following:concentrated for the relative density to 1.10(80C hot test),cold,add ethanol to the solution for the ethanol content of the solution reached 80%,static settlement for 24 hours. ConclusionThe extracting method is reasonable,stable,and suitable to industrialized producting.

AIM: To observe whether transfection of mammalian expression vector pEGFP containing the gene of B-cell specific moloney leukemia virus insertion site 1(BMI-1) could express in human cervix cancer cell line HeLa, and to detect its effect on HOX family expression and cell cycle.METHODS: pEGFP-BMI-1 was transfected into HeLa cells with Lipofectamine 2000. The expression of pEGFP-BMI-1 was determined by EGFP fluorescence and Western blotting. SYBR green I real-time RT-PCR was used to quantitate mRNA expression of P16~(INK4a), hTERT, HOXA9, HOXB4 and HOXC13. FACS analysis was used to detect the change of cell cycle.RESULTS: In HeLa cells transfected with pEGFP-BMI-1, the results of real-time RT-PCR showed that the mRNA expressions of P16~(INK4a), HOXA9 and HOXC13 were reduced to 9.2%, 10.9% and 69.7%, respectively, as compared to control HeLa cells (P<0.01). However, hTERT and HOXB4 mRNA expressions did not change significantly (P>0.05). FACS analysis showed a decrease from 65.68 % to 50.53% in G_1 population and a significant increase from 27.17% to 39.59 % in S population after transfection (P<0.01).CONCLUSION: BMI-1 over-expression in HeLa cells down-regulates mRNA expressions of P16~(INK4a), HOXA9 and HOXC13, decreases G_1 population and increases S population. Therefore, BMI-1 may be involved in carcinogenesis and cancer development.