HepG2 cells were incubated with dexamethasone,glucocorticoid receptor (GR) antagonist RU486,and pioglitazone for 24 or 48 h.Glucose concentration in culture medium was detected.The results showed that 10 μmol/L RU486 blocked the surge of glucose concentration by 48 h of dexamethasone treatment and the glucose concentration in RU486 + pioglitazone group was higher than pioglitazone group( P＜0.05 ),suggesting that the cross talk between PPARγ and GR may exist in hepatic glucose metabolism.

Objective To seek the ancients’ method for acupuncture treatment of epigastric pain and explore the ancients’ regularities in point selection for acupuncture treatment of this disease to provide a reference and basis for guidance of modern clinical treatment. Method Search words related to epigastric pain were established. A database was set up. Frequency analysis and data mining technique-associated rules were used to investigate the use frequencies of different acupoints and meridians, and regularities in acupoint combination in ancient acupuncture treatment of epigastric pain. Result In ancient acupuncture treatment of epigastric pain, the points more frequently used and more accepted were Zhongwan (CV12), Zusanli (ST36), Neiguan (PC6), Shangwan (CV13), Geshu (BL17), Gongsun (SP4) and Jianli (CV11). Meridian points were the main ones. The points of meridians going through the epigastrium were often selected. Acupoint combination was mainly based on syndrome differentiation-based point selection plus symptomatic point selection.

Objective To investigate the protective effects of trigonelline in neonatal rat cardiomyocytes during hypoxia /reoxygenation and its mechanism .Methods The neonatal rat cardiomyocytes was randomly divided into the normal control group , the hypoxia/reoxygenation group and the trigonelline group .Flow cytometry was used to determine the apoptosis and mitochondrial membrane potential .The levels of SOD and MDA were measured in different groups .Western blot method was used to measure the procaspase‐9 ,cleaved caspase‐9 ,procaspase‐3 and cleaved caspase‐3 protein level .Results Trigonelline could inhibit apoptosis (P<0 .05) ,enhanced activity of SOD (P<0 .05) ,reduce production of MDA (P<0 .05) ,stabilize the mitochondrial membrane potential (P<0 .05) and activate caspase‐9 and caspase‐3 in neonatal rat cardiomyocytes during hypoxia/reoxygenation .Conclusion Trigo‐nelline could protect myocardial cells from injury caused by hypoxia and reoxygenation ,and the mechanism may be associated with anti‐lipid peroxidation and stabilizing mitochondria membrane potential .

Objective To study the correlation between homocysteine levels and MTHFR C677T gene polymorphism of cerebral infarction.Methods Four hundred and fifty patients with cerebral infarction in our hospital were selected as the study group from February 2012 to August 2015,including 181 diabetes patients and 269 non-diabetic patients.Also 285 cases of physical examination healthy people in the outpatient department were selected as the control group.The MTHFR C677T polymorphism and the correlation between genotype and Hcy levels were analyzed by pyrosequencing.Results The difference of distributions of MTHFR genotype between the study group and the control group was statistically significant (P ＜ 0.05).The frequency of the T gene in the study group was significantly higher than the control group,the difference was statistically significant (x2 =13.67,P =0.00).The Hcy concentrations of the study group was significantly higher than the control group,and the difference was statistically significant (t =12.71,P =0.00).The Hcy levels of different MTHFR genotype in the cerebral infarction patients were statistically significant (F =17.68,P =0.00).Hcy levels in non-diabetic patients with cerebral infarction was significantly higher than in diabetic patients with cerebral infarction,and the difference was statistically significant (t =2.97,P =0.00).Hcy concentrations of TT genotype of non-diabetic group was significantly higher than the TT genotype of the diabetic group,CC type Hcy concentration significantly lower than the diabetic group,and the differences were statistically significant (t =5.67,2.18;P =0.00,0.03).In cerebral infarction patients both with non-diabetic and diabetic,the Hcy levels of MTHFR gene TT genotype were significantly higher than those of CC and CT genotype,and the differences were statistically significant (P ＜ 0.05),and the differences of Hcy levels between the genotype of CT and CC was not statistically significant (P ＞ 0.05).Conclusion The T allele frequency of MTHFR C677T in the cerebral infarction patients is much higher than healthy people.MTHFR TT genotype is related to serum Hcy levels.Maybe it is a risk factor for cerebral infarction.

A novel hemizygous frameshift mutation in exon1of DAX-1 gene (993delC) was found in a patient with late-onset adrenal hypoplasia congenita and hypogonadotropic hypogonadism.This mutation led the stop codon to appear in advance of 59 amino acids.His mother and two sisters were the carriers of this hemizygous mutation while his father and brother were wild-type.After glucocorticoid hormone replacement therapy, the clinical symptom was improved, but the level of ACTH was not suppressed.

BACKGROUND: Vascular endothelial growth factor (VEGF) can promote angiogenesis, and has been extensively used in treatment of bone defect. However, few studies have addressed its isomer VEGF121. OBJECTIVE: To construct adenovirus vector carrying VEGF121-FLAG and humanized Renilla reniformis green fluorescent protein 1(hrGFP-1) and observe its expression in bone marrow stromal stem cells (BMSCs). METHODS: Using polymerase chain reaction technique, VEGF121 gene contained in the plasmid of pTG19T-VEGF121 was used to remove termination codon. NotI and Xho I restriction sites were added before and after gene sequence. Obtained gene subclone was moved onto pMD19-T plasmid. The pMD19-T-VEGF121 and pShuttle-CMV-IRES-hrGFP-1 plasmids underwent double enzymatic digestion. Small fragment and big fragment were retrieved utilizing gel. Subsequently, coupled reaction was conducted to complete the construction of shuttle plasmid. After measuring virus titer, BMSCs were transfected and the fluorescence intensity was observed under fluorescence microscope.RESULTS AND CONCLUSION: Recombinant adenovirus plasmid was successfully constructed by enzymatic digestion determination and gene sequence. Fluorescence microscope has shown that BMSCs transfected with recombinant adenovirus presented significantly green fluorescence expression. Thus, adenovirus vector carrying VEGF121-FLAG and hrGFP-1 gene can express in eukaryotic cells, which can be used for gene therapy for ischemic disease.

BACKGROUND: Hypoxia-inducible factor-1 (HIF-1) can regulate the co-expression of various genes, and can induce angiogenesis with integrated physiological function.OBJECTIVE: To construct a novel adenoviral eukaryotic expression vector that can co-express mutant hypoxia-inducible factor-1 alpha (HIF-1a) target protein and humanized Renilla reniformis green fluorescent protein (hrGFP) reporter molecule under normoxic conditions.METHODS: The human HIF-1α gene carried by target gene donor plasmid pCMV6-XL5-HIF1α was sequenced and the site of restriction enzyme in above gene was analyzed. Site-directed mutagenesis of three amino acids including the 402 location, the 564 location, and the 803 location in gene coding region in HIF-1α were performed by polymerase chain reaction and sequencing was also done for monitoring mutation. The HIF-1α gene mutated correctly (HIF-1αmu) was coupled to adenoviral shuttle vector pShuttle-CMV-IRES- hrGFP-1. The recombinant adenovirus shuttle vector carrying HIF-1αmu gene was transferred to BJ5183-AD-1 electroporation competent cells after sequencing identification and Pme I restriction enzyme linearization.HIF-1αmu and hrGFP gene as well as hemeo-expression elements of hrGFP gene were reconstructed into adenoviral genome plasmids using homologous recombination mechanism in bacterium. Recombinants were obtained by Pac I restriction enzyme digestion and sequencing identification.RESULTS AND CONCLUSION: Amino acids including the 402 location, the 564 location and the 803 location in gene coding region in HIF-1α had become alanine after site-directed mutagenesis. Recombinant adenoviral expressing vector was successful as confirmed by restriction enzyme digestion and sequencing. These findings demonstrate that a novel recombinant adenoviral mutant eukaryotic expression vector pAd-HIF1αmu-IRES-hrGFP-1 was successfully constructed.

Employed staffs in military hospital are the basic power in the development of hospital, and they are playing an im-portant role in the military medical security.The article gives a detailed elaboration on the innovation and practice of employed staff man-agement in our hospital.In order to improve the efficiency and scientific management level of employed staffs, the innovative measure-ments in employment pattern, post management, education development, policy system, promotion passage are put forward in combination with current situation of employed staff in order to promote the construction of employed staffs and the development of the hospital.

OBJECTIVE:To explore the effects and safety of Hedan tablets on the levels of blood lipid,inflammatory factors and oxidative stress in coronary heart disease(CHD)patients with type 2 diabetes mellitus(DM2). METHODS:A total of 118 pa-tients diagnosed as CHD with DM2 were collected from our hospital during Jul. 2013-Nov. 2015,and then divided into observation group(63 cases)and control group(55 cases)according to random number table. Control group received conventional treatment such as controlling blood glucose,regulating blood lipid,vasodilator,anticoagulant. Observation group additionally received He-dan tablets 1.46 g before meal,tid,on the basis of control group. Both groups received treatment for 6 months. The levels of blood lipid (TC,TG,LDL-C,HDL-C),serum inflammatory factors (TNF-α ,IL-10,IL-6,hs-CRP) and oxidative stress indexes (MDA,SOD,NO)before and after treatment as well as the occurrence of ADR were compared in 2 groups. RESULTS:Before treatment,there was no statistical significance in levels of blood lipid,serum inflammatory factors or oxidative stress indexes be-tween 2 groups(P>0.05). After treatment,the serum levels of TC,TG,LDL-C,TNF-α,IL-6,hs-CRP and MDA in 2 groups were decreased significantly,while those of IL-10,SOD and NO were increased significantly;the observation group was signifi-cantly better than the control group,with statistical significance(P<0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Hedan tablets can effectively improve the level of blood lipid,reduce the lev-el of inflammatory factors,relieve inflammatory injury,reduce the level of oxygen free radical,strengthen antioxidant capacity and lighten oxidative stress injury with good safety.

Proteomic analysis is an effective way to identify protein constituent in Lewy bedy-like inclusions (or aggresome) in vitro. Exposure to synthetic proteasome inhibitor (PSI, 10 μmol/L) for 48 hours was used to induce the formation of cytoplasmic proteineous inclusions (termed as PSi-induced inclusions) in PC12 cells.The proteomic approaches of biochemical fractionation, two-dimensional electrophoresis (2-D) and identification via peptide mass fingerprints (PMF) were deployed, and 20 protein components of LBs were identified,i ncluding 2 proteins involved in the production of synaptic neurotransmitter, 6 subunits of the 26 S proteasome,2 cytoskeleton proteins, 2 subunits of mitochondrial complexes, 1 anti-oxidant protein, and 7 chaperone proteins and (or) chaperone-like proteins. The results suggested that these LB protein components might had been recruited in PSI-induced inclusions formed in PC12 cells under the condition of proteasome inhibition.