Objective To explore the immune mechanism of Tibetan medicine Twelve Flavors Yishou San in intervening with acute lung injury.Methods Construct a lipopolysaccharide(LPS)-induced acute lung injury rat model and treated it with Twelve Flavors Yishou San.The intervention effect of Twelve Flavors Yishou San on LPS induced acute lung injury was determined through lung pathological sections and blood gas analysis.Enzyme linked immunosorbent assay(ELISA)was used to detect the expression of pro-inflammatory cytokine tumor necrosis factor-α(TNF-α)and anti-inflammatory cytokine interleukin(IL)-10 in rat's bronchoalveolar lavage fluid.Flow cytometry was used to detect changes in immune cells in rat's bronchoalveolar lavage fluid,lungs,spleen and peripheral blood.Results LPS successfully induced an acute lung injury model in rats,and Twelve Flavors Yishou San could alleviate the exudation of inflammatory cells,elevated oxygen partial pressure in LPS induced acute lung injury to varying degrees.The ELISA results of rat bronchoalveolar lavage fluid showed that compared with normal control group,the TNF-α level in the bronchoalveolar lavage fluid of rats with acute lung injury was increased and the IL-10 level was decreased,while the Twelve Flavors Yishou San could alleviate this effect.The results of flow cytometry showed that Twelve Flavors Yishou San could significantly reduce neutrophil infiltration in rat bronchoalveolar lavage fluid,but had no significant effect on macrophages and adaptive immune response.Conclusion Twelve Flavors Yishou San can alleviate LPS induced acute lung injury by inhibiting neutrophil infiltration,providing an important experimental basis for fully understanding the therapeutic mechanism of Twelve Flavors Yishou San.

OBJECTIVETo construct and identify lentiviral vector containing human ILK-shRNA and mda7 gene.

METHODSBased on the human ILK gene sequences, RNAi target sequences were designed and cloned into the lentiviral vector pSicoR-eGFP by restriction endonuclease HpaI and XhoI double digestion and T4 DNA ligase ligation. Based on the human mda7 gene sequences, PCR primers were designed to clone the full-length mda7, and were cloned into the lentiviral vector pLVX-Puro. After the candidate clones were identified by DNA sequencing, the recombinant plasmid and the three packaging plasmids were co-transfected into the human embryonic kidney 293T cells by lipofectamine 2000 to produce the lentiviral particles. Human prostate cancer PC-3 cells were infected with the constructed lentiviral vector. The ILK and mda7 expression levels in PC-3 cells were quantified by qPCR and Western blot, respectively. The effect of ILK and mda7 on proliferation and migration of PC-3 cells were assessed by MTT method and Transwell assay, respectively.

RESULTSILK-pSicoR-eGFP and mda7-pLVX-Puro lentiviral vectors were successfully constructed. Strong green fluorescence was observed in the 293T cells under the fluorescent microscope after co-transfection of 293T cells with 4 plasmids of lentiviral vector. The transfection efficiency of the collected virus exceeded 90% in the 293T cells and the PC-3 cells were infected with the lentiviral particles with high efficiency. The A and B lentiviral vector inhibited the expression of ILK at both the mRNA and protein levels in PC-3 cells significantly. The mda7-pLVX-Puro lentiviral vector increased the expression of mda7 in PC-3 cells, and the ability was maintained for one month. Within 96 h, ILK and mad7 significantly inhibited the proliferation and migration of PC-3 cells (Ps<0.05).

CONCLUSIONThe lentiviral vectors of ILK knockdown and mda7 over-expression have been successfully constructed and identified. The recombinant lentivirus can efficiently infect human prostate cancer PC-3 cells, in which ILK expression is inhibited and mda7 is over-expressed.

Cell Line ; Genetic Vectors ; Humans ; Interleukins ; genetics ; Lentivirus ; genetics ; Plasmids ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo determine the risk factor of HCC in Guizhou.

METHODSA group case-control study design was conducted between 762 cases and 798 controls in Guizhou province. The main related-factors were analyzed with unconditional logistic regression model and evaluated by odds ratio (OR) and 95% confidence interval (95% CI).

RESULTSThere are significant differences between cases and controls in regarding to cigarette smoking 210 (27.6%),non-alcoholic fatty liver disease 336 (44.1%), alcoholic liver disease 245 (32.2%), family history of HCC 141 (16.5%), alcohol consumption 300 (39.4%), HBV infection 436 (57.2%), pickled food 290 (38.1%), and economic status 5 years ago 420 (55.1%) in cases,and cigarette smoking 116 (14.5%),non-alcoholic fatty liver disease 160 (20.1%), alcoholic liver disease 101 (12.7%), family history of HCC 40 (5.0%), alcohol consumption 180 (22.6%), HBV infection 82 (10.3%), pickled food 225 (28.2%), and economic status 5 years ago 647 (81.1%) in controls, with OR of each variable was 3.520, 2.464, 4.330, 2.219, 2.451, 19.245, 6.212, 0.174 respectively, P less than 0.01.

CONCLUSIONHBV infection and pickled food were the most common risks for HCC in Guizhou. Alcohol consumption excessively and cigarette smoking may increase the risk too.

Adult ; Aged ; Aged, 80 and over ; Alcohol Drinking ; Carcinoma, Hepatocellular ; epidemiology ; etiology ; Case-Control Studies ; China ; epidemiology ; Feeding Behavior ; Female ; Hepatitis B ; epidemiology ; Humans ; Liver Neoplasms ; epidemiology ; etiology ; Male ; Middle Aged ; Risk Factors ; Young Adult

OBJECTIVETo investigate the expression of HERC4 in human breast cancer tissues and its relationship with the clinicopathological features.

METHODSRT-qPCR was used to detect mRNA expression of HERC4, and Western blotting and immunohistochemistry were employed to detect protein expression of HERC4 in 67 breast cancer tissues and adjacent breast tissues.

RESULTSThe results of RT-qPCR showed a significantly higher mRNA expression of HERC4 in breast cancer tissues than in the adjacent breast tissues (P<0.05). Western blotting demonstrated HERC4 over-expression in breast cancer tissues compared with the adjacent breast tissues (P<0.05). Immunohistochemistry revealed HERC4 expression located predominantly in the cell cytoplasm. Positive HERC4 expression was detected in 61.2% of the breast cancer tissues as compared to 19.4% in the adjacent breast tissues, and its expression level was closely correlated with TNM stage and the histological grade (P<0.05).

CONCLUSIONHERC4 is correlated with the tumorigenesis and progression of breast cancer and may serve as a potential biomarker for early diagnosis and also as a potential therapeutic target in breast cancer.

Blotting, Western ; Breast ; metabolism ; Breast Neoplasms ; metabolism ; Disease Progression ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Real-Time Polymerase Chain Reaction ; Ubiquitin-Protein Ligases ; metabolism

OBJECTIVETo examine cerebral pathologies in cerebral amyloid angiopathy in a rat model of Alzheimer's disease.

METHODSRat models of Alzheimer's disease was established by stereotactic Aβ1-42 fiber injection in the bilateral hippocampus. The cognitive function of the rats was evaluated with water maze test. HE staining, Congo red staining and double-labeling indirect immunofluorescence were used to examine the dynamic distribution of Aβ fiber deposit in the brain.

RESULTSThe model rats showed significant differences from the control rats in the escape latency and the times of crossing platform in waster maze test. HE staining revealed a decreased number and degeneration of the granular cells with increased glial cells in the model rats. Congo Red staining showed that the Aβ fiber was deposited gradually in the small vessels in the brain parenchyma to cause thickening, stenosis or occlusion of the small vessels. Immunofluorescence staining detected Aβ fiber migration from the parenchyma to the walls of the small arteries in the rat models.

CONCLUSIONCerebral amyloid angiopathy is a major pathological feature in Alzheimer's disease.

Alzheimer Disease ; pathology ; Amyloid beta-Peptides ; chemistry ; Animals ; Brain ; pathology ; Cerebral Amyloid Angiopathy ; pathology ; Disease Models, Animal ; Rats ; Staining and Labeling

Objective To examine cerebral pathologies in cerebral amyloid angiopathy in a rat model of Alzheimer's disease. Methods Rat models of Alzheimer's disease was established by stereotactic Aβ1-42 fiber injection in the bilateral hippocampus. The cognitive function of the rats was evaluated with water maze test. HE staining, Congo red staining and double-labeling indirect immunofluorescence were used to examine the dynamic distribution of Aβ fiber deposit in the brain. Results The model rats showed significant differences from the control rats in the escape latency and the times of crossing platform in waster maze test. HE staining revealed a decreased number and degeneration of the granular cells with increased glial cells in the model rats. Congo Red staining showed that the Aβ fiber was deposited gradually in the small vessels in the brain parenchyma to cause thickening, stenosis or occlusion of the small vessels. Immunofluorescence staining detected Aβ fiber migration from the parenchyma to the walls of the small arteries in the rat models. Conclusion Cerebral amyloid angiopathy is a major pathological feature in Alzheimer's disease.

Objective To examine cerebral pathologies in cerebral amyloid angiopathy in a rat model of Alzheimer's disease. Methods Rat models of Alzheimer's disease was established by stereotactic Aβ1-42 fiber injection in the bilateral hippocampus. The cognitive function of the rats was evaluated with water maze test. HE staining, Congo red staining and double-labeling indirect immunofluorescence were used to examine the dynamic distribution of Aβ fiber deposit in the brain. Results The model rats showed significant differences from the control rats in the escape latency and the times of crossing platform in waster maze test. HE staining revealed a decreased number and degeneration of the granular cells with increased glial cells in the model rats. Congo Red staining showed that the Aβ fiber was deposited gradually in the small vessels in the brain parenchyma to cause thickening, stenosis or occlusion of the small vessels. Immunofluorescence staining detected Aβ fiber migration from the parenchyma to the walls of the small arteries in the rat models. Conclusion Cerebral amyloid angiopathy is a major pathological feature in Alzheimer's disease.

Objective To learn the current obstetric beds allocation,obstetric service delivery and access in midwifery institutions at various levels in the localities and nationwide as well as the trends,and assess the equity of obstetric beds allocation,for the purpose of recommending on a reasonable adjustment of obstetric beds allocation in these institutions.Methods Random sampling made in 44 cities in China for questionnaire survey on obstetric beds allocation,service delivery and utilization in 2008～2010 at all midwifery institutions in these cities.Interview of health administrators and medical workers at obstetric of midwifery institutions in 6 counties.Results Every thousand square kilometers in China average 5.9 midwifery institutions,and every thousand people average 0.24 obstetric beds.During 2008 ～ 2010,the number of beds and midwifery service delivery kept a growing trend while in townships obstetric service deliveries maintain a falling trend.The fairness index of maternity beds layout nationwide is 0.021283.Obstetric beds supply and demand ratio in different regions ranges among 0.97～ 1.41.The number of deliveries made in county level hospitals is 54.1％,but the beds in such hospitals account for but 48.1％.Conclusion The equity of obstetric beds allocation is good in terms of population layout.The quantity of obstetric beds meets the current needs nationwide,while the allocation of obstetric beds is less than reasonable among obstetric institutions of various levels in the country.The authors recommend a better control and adjustment of midwifery institutions layout,especially more such hospitals at the township level to meet the need of women delivery in hospital in remote areas.

ObjectiveTo observe the efficacy and safety of inhaled tiotropium capsule on adult bronchial asthma.MethodsThirty-four adult asthma patients were randomized into two groups.We evaluated the addition of tiotropium bromide to an inhaled glucocorticoid in the treatment group( n =17 ),as compared with the addition of the LABA Formoterol in the control group ( n =17 ).Serial measurements of lung function,symptom control and quality of life were performed.ResultsAfter 8 weeks of treatment,peak expiratory flow (PEF) in the addition of tiotropium bromide group[ ( 359 ± 12 ) L/min vs.( 275 ± 5 ) L/min,P ＜ 0.05 ] and the addition of the LABA Formoterol group [ ( 346 ± 11 ) L/min vs.( 275 ± 9 ) L/min,P ＜ 0.05 ] were significantly higher than those before treatment.The forced expiratory volumes in one second ( FEV1 )were also significantly elevated in both groups [ Tiotropium bromide group:( 2.80 ± 0.28 ) L vs.( 2.30 ± 0.28 ) L,P ＜ 0.05 ; LABA Formoterol group:( 2.69 ± 0.34 ) L vs.( 2.25 ± 0.34 ),P ＜ 0.05 ] compared with pre-treatment.Average dose of emergent drug ( ventolin ) usage were decreased in both groups after treatment [ Tiotropium bromide group:(0.96 ± 0.34 ) puff/d vs.(4.11 ± 1.03 ) puff/d,t =3.05,P ＜ 0.05 ; LABA Formoterol group:( 0.88 ± 0.44 )puff/day vs.( 4.43 ± 0.87 ) puff/day,t =3.23,P ＜ 0.05 ].Meanwhile,significantly improved quality of life scores and alleviation of asthma symptom were found in both groups as compared with baseline ( P ＜ 0.05).However,no significant differences in the above variables after treatment were found between two groups( P ＞0.05 ).ConclusionWhen added to an inhaled glucocorticoid,tiotropium improved symptoms and lung function in patients with inadequately controlled asthma.Its effects appeared to be equivalent to the addition of Formoterol.

Objective To identify one umbilical blood sample with abnonnal gap-PCR products of three bands of α2,-α3.7 and-SEA.further family pedigree were analyzed for the source of genetic variations,Methods One fetal umbilical blood sample was drawn from a woman of 24-weeks pregnancy.Gap-PCR for α-thalassemia was routinely conducted and abnornlal three bands of α2.-α3.7 and-SEA were observed.which could not be interpreted according to the kit manual and suspected as rare variation.With informed consen,DNA samples from the parents and grandparents were obtained for further study.Singleplex andnested PCR techniques were utilized to analyze the molecular characteristics of DNA samples from this fetus and its parents and grand-parents.Results Hematological phenotype study showed that fetal Hb Ban's was 7.6%,and its mother and maternal grandfather were both with typical α-thalassemia.while its father,grandfather and grandmother and maternal grandmother are without abnormal hematological change.Molecular study showed that fetal blood DNA was a heterozygosity for HKaa and-SEA.its father and grandfather are both HKαα/αα,its mother and maternal grandfather are both-SEA/aa,its grandmother and maternal grandmother are with both normal alleles of αα/αα.Then after genetic counseling the fetas was saved and iS a she baby now.Conclusion ThroUgh careful molecular tests one case of prenatal heterozygosity of HKαα/-SEA was identified,and the fetus is kept successfully through careful clinical counseling.