There have been very few studies on the effect of Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction in inhibiting apoptosis in myocardial ischemial injury caused by coronary heart disease. In this experiment, Gualou Xiebai Banxia decoction combined with-Xuefu Zhuyu decoction were used to intervene the miniature swine phlegm and blood stasis type coronary heart disease model, in order to observe the effect of the combined prescription on the myocardial apoptosis and the expressions of Bcl-2, Bax, Caspase-3, Caspase-9 in the model. Totally 15 Chinese experimental miniature swine were adopted and randomly divided into the control group, the model group and the phlegm and stasis-treating group. The model group and the stasis-treating group were fed with high fat diets for two weeks, intervened with the coronary artery injury and then given drugs and high fat diets for eight weeks. The control group was fed with ordinary diets for 10 weeks, without the coronary artery injury. After the experiment, myocardia at the juncture of infracted areas were collected and made into formalin-fixed paraffin sections. The TDT-mediate dUTP nick end labeling (TUNEL) assay was used to detect the myocardial apoptosis. The immunohistochemistry (IHC) technique was applied to detect Bcl-2, Bax, Caspase-3, Caspase-9 levels in myocardial tissues. According to the findings, the apoptosis indexes (AI) for the control group, the model group and the phlegm and stasis-treating group were 0.92%, 27.68%, 17.28%, respectively. The AI of the phlegm and stasis-treating group was significantly lower than that of the model group (P < 0.01). Compared with the model group, the phlegm and stasis-treating group showed significantly higher Bcl-2 protein expression (P < 0.01) and lower Bax, Caspase-3 and Caspase-9 protein expressions (P < 0.01). In conclusion, Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction have a significant protective effect against the myocardial apoptosis in miniature swine phlegm and blood stasis type coronary heart disease model.
Animals ; Apoptosis ; drug effects ; Caspases ; metabolism ; Coronary Disease ; drug therapy ; Disease Models, Animal ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Male ; Medicine, Chinese Traditional ; Myocardium ; pathology ; Phytotherapy ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Swine ; Swine, Miniature ; bcl-2-Associated X Protein ; analysis

OBJECTIVETo explore whether there is a relationship between gross tumor volume (GTV) and pathologic lymph node metastasis or prognosis of esophageal carcinoma, and to provide a new prognosis reference for esophageal carcinoma (EC).

METHODSSix hundred and seven patients received radical resection of thoracic esophageal carcinoma from May 2002 to June 2006 in our hospital, and their pre-operative CT images were transmitted to the three-dimensional conformal radiotherapy planning system by the network in digital format. Esophageal GTV targets were outlined and their GTV volumes were calculated. To analyze whether there is a relationship between GTV volume and pathologic lymph node metastasis or prognosis.

RESULTSIn the 607 cases of esophageal carcinoma, the GTV volume was (22.5 ± 16.8) cm(3) in 374 stage N0 EC patients, significantly different from that of (30.4 ± 20.1) cm(3) in 233 stage N1 EC cases (P < 0.001). There is a significant difference between the GTV volumes of the groups with and without lymph node metastasis (P < 0.05). There was a significant difference of the GTV volumes of EC patients with one lymph node metastasis and those with ≥ 4 lymph node metastasis (P < 0.05). There was a positive correlation between GTV volume and the number of lymph node metastasis (r = 0.230, P < 0.001). The 1-, 3-, 5-year survival rates since the surgery date were 83.8%, 53.5%, and 36.4%, respectively. There was a significant difference between the survival rates of stage N0 (48.5%) and stage N1 patients (18.2%, P < 0.001), and there was a significant difference between the survival rats of patients with 0, 1 and ≥ 2 lymph node metastasis (P < 0.01). Cox regression model analysis showed that GTV volume, number of lymph node metastasis, pathological type, and lesion site were independent prognostic factors (all P < 0.05).

CONCLUSIONThe GTV volume of esophageal carcinoma is positively correlated with the number of pathologic lymph node metastasis, and it is an independent prognostic factor for this cancer.

Adenocarcinoma ; diagnostic imaging ; pathology ; surgery ; Adult ; Aged ; Carcinoma, Small Cell ; diagnostic imaging ; pathology ; surgery ; Carcinoma, Squamous Cell ; diagnostic imaging ; pathology ; surgery ; Esophageal Neoplasms ; diagnostic imaging ; pathology ; surgery ; Esophagectomy ; methods ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Proportional Hazards Models ; Radiotherapy, Conformal ; Survival Rate ; Tomography, X-Ray Computed ; Tumor Burden

The capsid protein (VP60) gene of RHDV was subcloned into the Pichia expressin vector pPICZ B to express the VP60 protein intracellularly. The recombinant plasmid was initially transformed into a E. coli strain TOP10 F'. After verification of the construct by sequencing, the recombinant plasmid was linearized by Sac I in the 5' AOX1 region and then transformed into Pichia pastoris strain GS115 using the Pichia EasyComp Kit. After selecting and verifing for the insertion of VP60 gene in the genome, two clones of Pichia transformants were select for expression test. The recombinant clones were first inoculate with BMGY in baffled flask at 28-30 degrees C in a shaking incubator (250-300 r/min) until culture reaches an OD600 = 2-6, then resuspend the cell pellet to an OD6oo of 1.0 in BMMY medium to induce expression for 5 days by methanol at a concentration of 0.5% in a 1 liter baffled flask covered with 2 layers of sterile gauze. Collect the cell pellets and break it by acid-washed 0.5 mm glass beads. The expression of recombinant Pichia strains was detected by SDS-PAGE and Western analysis with a polyclonal serum which showed a specific protein band of 60kD. Theses results indicates that the recombinant VP60 produced in Pichia was antigenically similar to the viral polypeptide. Electron microscopic observation of the recombinant Pichia-derived protein revealed the presence of virus-like particles similar in size and appearance to native virus capsids. In the haemagglutination test, the recombinant VLPs, like the native RHDV, also agglutinated human blood type O erythrocytes and could be inhibited by the anti-RHDV polyclonal serum.
Animals ; Capsid ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; metabolism ; Hemorrhagic Disease Virus, Rabbit ; genetics ; Pichia ; genetics ; metabolism ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Viral Structural Proteins ; biosynthesis ; genetics ; isolation & purification

AIM:To explore the molecular mechanism of panaxadiol saponin(PDS)by observing Toll like receptor(TLR)2 and TLR9 mRNA expression induced by lipopolysaccharide(LPS).METHODS:Rats were divided into LPS,LPS+PDSL,LPS+PDSM and control group,respectively.Nitric oxide synthase(NOS)activity,nitric oxide(NO)content,LPO content,SOD activity and TLR2 and TLR9 mRNA expression were assayed 4 h after intravenous injection of LPS.RESULTS:NOS activity,NO content,LPO content of LPS+PDSL group and LPS+PDSM group were significantly lower than those in LPS group.TLR2 mRNA expression in the liver tissue of LPS+PDSL group and LPS+PDSM group was decreased compared with LPS group.CONCLUSION:PDS has a protective effect on liver tissues by triggering the down-regulation of TLR2 expression,reducing NOS activity,and NO content.

Objective To observe the therapeutic effect of the sacral nerve stimulation (SNS)on the neurogenic bladder. Methods SNS was used to treat 94 patients with neurogenic bladder. The ICI-Q-SF scores, voiding diary (including urinary incontinence times, urinal pad test, urination times,nocturia times and urinary volume) and urine dynamics test (including bladder volume, pressure of detrusor, maximum urinary flow rate, average urinary flow rate, pressure of bladder neck, maximal urethral pressure, functional urethral length and residual urine volume) were observed before and 1 week after SNS. And the data was analyzed statistically. Results The ICI-Q-SF scores were 17.2±1.8 before the treatment of SNS and 8.3±1.6 after SNS (P＜0.05). The urinary incontinence times, urination times and nocturia times were significantly decreased (P＜0.05) compared with those before treatment. The weight of urinal pads was lessened significantly (P＜0.05), while the urinary volumes were increased significantly (P＜0.05). The bladder volume, the maximum urinary flow rate and the average urinary flow rate were increased significantly (P＜0.05) while the residual urine volumes were decreased significantly (P＜0.05). There were no differences in pressure of detrusor, pressure of bladder neck, maximal urethral pressure and functional urethral length (P＞0.05). No complication occurred in our study and total effective rate was 75.5%. Conclusions The therapeutic effect of SNS on the neurogenic bladder is sure, and the symptom of the neurogenic bladder is well improved with few complications. It should be widely used.

Prolongation of the duration of heart preservation in vitro is very important in clinical heart transplantation. Previous studies have shown that mitochondrial ATP-sensitive potassium channel (mitoK(ATP)) plays an important role in cardioprotective effect. The purpose of this study was to assess whether the mitoK(ATP) opener diazoxide as an additive to cardioplegia solution could enhance myocardial protection during long-term hypothermic preservation of the rat heart. Langendorff model of isolated rat heart was used. After 30 min stabilization of perfusion, the hearts were stored in Celsior cardioplegia solution at 4 degrees C with (15, 30 and 45 micromol/L) or without diazoxide, a mitoK(ATP) channel opener, for 10 h followed by 60 min reperfusion. The recovery of cardiac contractile function, myocardial enzyme leakage in the coronary effluent, and myocardial water content were determined. The myocardial ultrastructure was also observed. We found that: (1) Diazoxide treatment improved the recovery of left ventricular developed pressure and +/-dp/dt(max) dose-dependently. Left ventricular end-diastolic pressure was significantly lower in diazoxide-treated hearts than that of hearts in Celsior solution after hypothermic preservation for 10 h. (2) Diazoxide at 30 and 45 micromol/L significantly decreased the water content of myocardium and increased coronary flow of the hearts compared to those in control. (3) The leakage of myocardial enzymes (lactate dehydrogenase, creatine kinase and glutamate-oxaloacetate transaminase) in the coronary effluent was significantly reduced in diazoxide-treated hearts. (4) Impairment of myocardial ultrastructure after 10 h hypothermic preservation was alleviated in hearts treated with 30 micromol/L diazoxide. (5) The cardiac effects of 30 micromol/L diazoxide were attenuated by a mitoK(ATP) blocker 5-hydroxydecanoate (5-HD, 100 micromol/L). These results indicate that diazoxide as a supplementation in cardioplegia solution could enhance myocardial protection during long-term hypothermic heart preservation via opening of mitochondrial K(ATP) channel.
Animals ; Cryopreservation ; Diazoxide ; pharmacology ; Heart ; In Vitro Techniques ; Male ; Mitochondria, Heart ; metabolism ; Organ Preservation Solutions ; pharmacology ; Potassium Channels ; metabolism ; Rats ; Rats, Sprague-Dawley ; Time Factors

OBJECTIVETo investigate whether the mitochondrial ATP-sensitive potassium channel (mitoK(ATP)) opener diazoxide as an additive to cardioplegia solution could enhance myocardial protection during hypothermic preservation of the rat heart.

METHODSThe Langendorff model of isolated rat heart was used. After equilibrium, the hearts were stored in Celsior cardioplegia solution at 4 degree with or without supplement of diazoxide for 3 or 8 h followed by 60 minutes reperfusion. The recovery of cardiac contractile function, myocardial enzyme leakage in the coronary effluent, and myocardial water content were determined. The myocardial ultrastructure was also observed.

RESULT(1) Treatment of diazoxide improved the recovery of left ventricular developed pressure and decreased the leakage of myocardial enzymes, lactate dehydrogenase (LDH) and creatine kinase (CK), at the 2nd and 4th minute of reperfusion of rat heart after hypothermic preservation for 3 h. (2) After hypothermic preservation for 8 h, diazoxide improved the recovery of left ventricular developed pressure and decreased the leakage of myocardial enzymes (LDH, CK and glutamic oxalic transaminase) during reperfusion. Moreover, left ventricular end-diastolic pressure was significantly lower in diazoxide-treated hearts than that of hearts in Celsior solution. (3) Diazoxide significantly decreased the water content of myocardium and increased coronary flow of the hearts compared with those in control after hypothermic preservation for 8 h. (4) Impairment of myocardial ultrastructure after 8 h hypothermic preservation was alleviated in hearts treated with 30 mol/L diazoxide. (5) The cardiac effects of 30 mol/L diazoxide were attenuated by a mitoK(ATP) blocker 5-hydroxydecanoate (100 micromol/L).

CONCLUSIONDiazoxide as a supplementation in cardioplegia solution could enhance myocardial protection during hypothermic heart preservation via opening of mitochondrial K(ATP) channel.

Animals ; Cardioplegic Solutions ; Cryopreservation ; Diazoxide ; pharmacology ; Heart ; Male ; Organ Preservation ; Organ Preservation Solutions ; pharmacology ; Potassium Channels ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVESome research has shown that hyperbaric oxygen (HBO) can decrease the rate of mortality and disability caused by hypoxic-ischemic encephalopathy (HIE) in neonates. However, the HBO pressure used in the clinical reports and the efficacy of HBO are different. This study was designed to investigate the efficacy of HBO therapy under different pressures by observing the changes of peroxidation, antioxidant levels and brain vasomotor regulation factors as well as the score of neonatal behavioral neurological assessment (NBNA) in neonates with HIE after HBO therapy.

METHODSSixty neonates with HIE were randomly administered with 1.4, 1.5 or 1.6 atmosphere absolute (ATA) of HBO, once daily for seven days. Serum levels of malondialdehyde (MDA), superoxide dismutase (SOD), nitric oxide (NO) and nitric oxide synthase (NOS) were measured before and after HBO therapy. Meanwhile, NBNA and eye ground examination were performed.

RESULTSSerum SOD level increased and serum levels of MDA, NO and NOS decreased significantly after HBO therapy in the three HBO therapy groups (P<0.01). Serum SOD level was significantly higher and serum levels of MDA, NO and NOS were significantly lower in the 1.6 ATA HBO group than those in the 1.4 ATA group after therapy (P<0.05). The 1.6 ATA HBO group also showed increased SOD and decreased MDA levels compared with the 1.5 ATA HBO group after therapy (P<0.05). NBNA scores in the three groups increased significantly after HBO therapy (P<0.05). None of the three HBO therapy group patients showed abnormal eye grounds after therapy.

CONCLUSIONSHBO therapy with 1.4, 1.5 or 1.6 ATA is safe and effective for neonatal HIE. The antioxidant capacity increases with the increasing HBO pressure in neonates with HIE.

Female ; Humans ; Hyperbaric Oxygenation ; Hypoxia-Ischemia, Brain ; drug therapy ; physiopathology ; Infant, Newborn ; Male ; Malondialdehyde ; blood ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; Pressure ; Superoxide Dismutase ; blood

OBJECTIVEIt was to explore the application of the hepatitis B virus large proteins (HBV-LP) in public health physical examination through clinical testing of HBV-LP in Catering practitioners, students and soldiers of New requisition etc.

METHODS709 samples of HBV infection people were collected in public health physical examination and all collected samples were detected. ELISA was used to detect HBV serum markers (HBV-M) and HBV-LP. Real-time PCR was applied to detect quantity of HBV DNA.

RESULTS(1) The detectable results of HBV-LP and HBV DNA were no statistical significance in 709 samples. (2) The detectable results of HBV-LP and HBV DNA were also no statistical significance in different models of HBV infection patients. The values of chi2 and P were respectively 2.67, 0.60, 0.00 and 0.10, 0.44, 1.00. S/A of HBV-LP and the copies of HBV-DNA had a positive relationship (r = 0.959, P < 0.05).

CONCLUSIONHBV-LP could reflect the level of hepatitis B virus replication and be used to judge the level of hepatitis B virus replication in people of public health physical examination. HBV-LP may be worth using widely in physical examination.

Adolescent ; Adult ; Child ; Female ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Liver Function Tests ; Male ; Middle Aged ; Public Health ; Viral Proteins ; genetics ; metabolism ; Young Adult

Host genetic factors, such as human leukocyte antigen (HLA) alleles, are important in Human immunod-eficiency virus (HIV) infection and its progression to AIDS. HLA class I genes, especially highly polymorphicHLA-B genes, are involved in the activation of HLA-restricted cytotoxic T lymphocytes (CTLs) against HIV, andthus control susceptibility to or protect against this virus. The present study was aimed to determine the distributionof HLA-B alleles in the Chinese Uygur ethnic group and its association with HIV infection. One hundred ten healthycontrol (HIV negative) and 128 HIV positive Chinese Xinjiang Uygur ethnic individuals were used in this study.HLA typing for B allele was performed by polymerase chain reaction (PCR) with sequence-specific primers (SSP).Hardy-Weinberg equilibrium was calculated using POPGENE software for the healthy control group. The HLA-Bfrequency of each allele was compared between the patients and the controls using the chi-square test. In HIV-1-pos-itive group, gene frequency of allele B * 4901 was significantly higher compared to the healthy control subjects (P=0.02, OR=3.06, 95%CI=1.16～8.10 forB*4901). In contrast, the gene frequency of B * 40 in healthy controlswas significantly higher than in the HIV-positive patients (P=0.02, OR=0.39, 95%CI=0.07～0. 92 for B* 40).In this study, HLA allele B * 4901 may be associated with increased susceptibility to HIV-1 infection, whereas the B* 40 allele may be associated with resistance to H HIV-1 infection.