To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Drug Synergism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; pathology ; Real-Time Polymerase Chain Reaction ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Transfection

OBJECTIVETo evaluate the relationship between mortality and HBVDNA and HBeAg expression of severe hepatitis B patients.

METHODSThe mortality rates of different types of severe hepatitis patients in our hospital during the last five years were analysed. HBV DNA was detected using the fluorescence quantitative PCR method and the HBeAg expression of severe hepatitis B was studied using a microparticle method.

RESULTS(1) Hepatitis B morbidity was 83.5% in each type of severe hepatitis, and severe chronic hepatitis B morbidity was 96.77% in each type of severe chronic hepatitis. (2) The mortality rate of those with HBV DNA more than 1 x 10(5) copies/ml was 53.25% and the mortality of those with HBV DNA less than 1 x 10(5) copies/ml was 34.50% (P less than 0.01). The HBeAg expression had no influence on the death rate. (3) The death rate descended to 30.38% from 54.64% (HBV DNA more than 1 x 10(5) copies/ml) when treated with Lamivudine (P less than 0.01).

CONCLUSIONIn severe hepatitis the quantity of virus carried in the patient is one of the key factors of mortality; antivirus treatment can lower mortality.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; DNA, Viral ; Female ; Hepatitis B Surface Antigens ; Hepatitis B virus ; genetics ; immunology ; physiology ; Hepatitis B, Chronic ; diagnosis ; virology ; Humans ; Male ; Middle Aged ; Viral Load ; Young Adult

Objective To observe the effect of electroacupuncture(EA)on the expression of NKCC1,KCC2 and activation of microglia in spinal dorsal horn of paclitaxel(PTX)-induced neuropathic pain rats and its possible mechanism.Methods Male SD rats were randomly divided into the vehicle group(vehicle),the PTX group,the PTX+EA group and the PTX+sham EA group,with 12 rats in each group.The rat model of PTX-induced neuropathic pain was established by intraperitoneal injection of PTX.After modeling,EA was applied to"Zusanli"and"Yanglingquan"for 7 days in the PTX+EA group.Paw withdrawal threshold and paw withdrawal latency were tested at 2 days before and 1,3,5,7,14 and 21 days after PTX injection.Immunofluorescence and Western blot assay were used to detect expression levels of sodium-potassium-chloride cotransporter 1(NKCC1),potassium-chloride cotransporters 2(KCC2)and microglia markers-ionized calcium binding adapter molecule 1(Iba1)in spinal dorsal horn.Results Compared with the vehicle group,mechanical and thermal hyperalgesia of both hind feet were found in the PTX group,and the expression of NKCC1 and the number of activated microglia in dorsal horn tissue of spinal cord were increased.Compared with the PTX group,mechanical and thermal hyperalgesia were significantly improved in the PTX+EA group at day 14 and 21,and the expression levels of NKCC1 and Iba1 in dorsal horn tissue of spinal cord were decreased.There was no significant difference in KCC2 expression between the four groups.Conclusion Electroacupuncture can effectively relieve paclitaxol-induced neuropathic pain,which may be related to the inhibition of NKCC1 expression and microglia activation in spinal dorsal horn of rats.

BACKGROUNDThe percutaneous transcatheter closure of secundum atrial septal defect (ASD) is increasingly a widespread alternative to surgical closure. The aim of this study was to assess long-term results of percutaneous closure of secundum-type atrial septal defect (ASDII).

METHODSBetween January 2001 and December 2005, 61 patients underwent a successful percutaneous closure of ASDII; including 25 male and 36 female. All were included in the patient study and were followed up to monitor by electrocardiogram and echocardiography, at intervals of 3 days, 3 months, 6 months, 1 year, 2 years, and 5 years after operation.

RESULTSThree days after percutaneous transcatheter septal closure (PTSC), the right atrium diameter, right ventricular end-diastolic left-right diameter and right ventricular end-diastolic volume (RVEDV) decreased significantly (P < 0.05). Right ventricular end-diastolic anteroposterior diameter (RVEDD), right ventricular end-systolic volume (RVESV) and right ventricular ejection fraction (RVEF) also decreased (P < 0.01). During the period from 3 to 6 months, the size of the right atrium and right ventricle returned to normal range. Three days after PTSC, the left ventricular end-diastolic diameter (LVEDD), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular-systolic volume (LVSV) and left ventricular ejection fraction (LVEF) were significantly increased (P < 0.05). At 1 year, the size of the left atrium, left ventricle and left cardiac function returned to normal range (P < 0.01). There were no deaths or significant complications during the study. At five year follow-up, all defects were completely closed and remained closed thereafter.

CONCLUSIONTranscatheter closure of ASDII effectively eliminated the abnormal shunt and, subsequently improved the dimensions of each chamber and cardiac function.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Heart Septal Defects, Atrial ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Ultrasonography ; Young Adult

OBJECTIVETo contrast single and double balloon-inflated kyphoplasty for vertebral compression fractures (VCFs) and evaluate its clinical efficacy.

METHODSFrom May 2000 to May 2004, 90 consecutive procedures were performed in 58 patients who suffered from painful vertebral compression fractures, transferring tumour and angioma. Ninety vertebrae were inflated while 62 as A group were double balloon and 28 as B group were single balloon, fracture reduction and bone cement augmentation. Preoperative and postoperative symptom levels, variables, complications were recorded and the vertebral height and Cobb angle were measured and analyzed.

RESULTSAll patients' pain was alleviated or disappeared without syndrome, and the vertebral height and Cobb angle of both groups were improved. The average recovery rate was 72.6% (22.9% approximately 100%), Cobb angle from 17.9 degrees (3.1 degrees approximately 31.6 degrees ) were corrected to 9.6 degrees (0.6 degrees approximately 28.2 degrees ), the average angle was 8.7 degrees (0.3 degrees approximately 27.2 degrees ), and the contrast between preoperative and postoperative showed obvious differences (P <0.001). The average recovery rate of A group was 77.6% (55.3% approximately 100%), B group was 64.3% (22.9% approximately 100%). The average postoperative Cobb angle of A group was 9.9 degrees (0.3 degrees approximately 27.2 degrees ), B group was 8.6 degrees (0.6 degrees approximately 19.8 degrees ) (P >0.05).

CONCLUSIONSAs a promising minimally invasive surgery, balloon kyphoplasty can provide early relief of pain and improve the function as well as spinal alignment in treatment of painful compression fracture owing to recovering the vertebral height and Cobb angle of the vertebral body. Single balloon-inflated kyphoplasty can improve VCFs as double balloon.

Adult ; Aged ; Aged, 80 and over ; Bone Cements ; therapeutic use ; Female ; Fractures, Compression ; complications ; surgery ; Humans ; Kyphosis ; etiology ; surgery ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Orthopedic Procedures ; instrumentation ; methods ; Retrospective Studies ; Spinal Fractures ; complications ; surgery ; Thoracic Vertebrae ; injuries ; surgery

0.05 ). After once injection both observers considered the number of clearly recognized endocardial border segments increased significantly. The number evaluated by observers A increased from 2.68 ? 0.95 to 5.99 ? 0.10 while from 2.82 ? 1.03 to 5.99 ? 0.11 by observers B( P 0.05 ). The average contrast enhancement rate of LV endocardial border was 99.7 %. Perfluoropropane-albumin microsphere injection had no significant effection on vital signs such as blood prssure, heart rate and respiration. Electrocardiogram didn′t change markedly and the variance of the laboratory findings like blood and urine routine examination, hepatic and renal function was in normal range. Only one case( 0.33 %) had slight side-effects who suffered from mild nausea and diarrhea, which suggested the clinical safety of this contrast agent. Conclusions Perfluoropropane-albumin microsphere injection could enhance the resolution of LV endocardial borders and make the judgement of regional myocardial movement easier. It has little side-effects and will be appropriate for clinical use.

Objective To investigate whether Heshouwuyin can delay the aging of Leydig cells in rat testis through DNA methyltransferase 1(DNMT1).Methods Totally 40 male Wistar rats were randomly divided into 4 groups,with 10 rats in each group.Immunohistochemistry was used to detect the expression levels of DNMT1 in testis tissue of rats.Testosterone content in serum of rats in each group was detected by ELISA test.A rat Leydig cell aging model was established by free radical oxidative damage.DNMT1 was knocked down by lentivirus in Leydig cells,and the cell senescence status and the testosterone content and testosterone synthesis key enzyme 3β-hydroxysteroid dehydrogenase(3β-HSD),cytochrome P450 family member 11A1(CYP11A1)content secreted by cells were detected by β-galactosidase(β-GAL)staining,immunofluorescenct staining and ELISA.Results Compared with the young control group(YCG),the expression of P16 protein and the positive rate of β-GAL in the testis tissue of rats in the natural aging group(NAG)increased significantly,and the expression of DNMT1 and serum testosterone levels decreased(P＜0.05).However,after Heshouwuyin intervention,the expression of P16 protein and the positive rate of β-GAL in the testis of aging rats were reduced,and DNMT1 expression and the serum testosterone levels increased(P＜0.05).The same trend was observed in Leydig cells.Knockdown of DNMT1 in Leydig cells,β-GAL positivity and P16 protein expression increased significantly,and testosterone secretion and testosterone synthesis key enzymes 3β-HSD,CYP11A1 content from Leydig cells decreased significantly,compared with the normal control group(NCG)(P＜0.05).When Heshouwuyin was added,the above phenomenon was improved.Conclusion Heshouwuyin can delay the aging of rat Leydig cells through DNMT1.

OBJECTIVETo examine whether the polymorphisms of endothelial nitric oxide synthase (eNOS) gene are associated with the susceptibility to high altitude pulmonary edema (HAPE) in Chinese railway construction workers at Qinghai-Tibet where the altitude is over 4 500 m above sea level.

METHODSA case-control study was conducted including 149 HAPE patients in the construction workers and 160 healthy controls randomly recruited from their co-workers, matching the patients in ethnicity, age, sex, lifestyle, and working conditions. Three polymorphisms of eNOS gene, T-786C in promoter, 894G/T in exon 7, and 27bp variable number tandem repeat (VNTR) in intron 4, were genotyped using polymerase chain reaction (PCR) and confirmed with DNA sequencing.

RESULTSThe frequencies of 894T allele and heterozygous G/T of the 894G/T variant were significantly higher in HAPE patients group than in the control group (P=0.0028 and P=0.0047, respectively). However, the frequencies of the T-786C in promoter and the 27bp VNTR in intron 4 were not significantly different between the two groups. Haplotypic analysis revealed that the frequencies of two haplotypes (H3,T-T-b, b indicates 5 repeats of 27 bp VNTR; H6, C-G-a, a indicates 4 repeats of 27 bp VNTR) were significantly higher in HAPE patients (both Pü0.0001). On the contrary, the frequencies of H1 (T-G-b) and H2 (T-G-a) were lower in HAPE patients than in healthy controls (both Pü0.001).

CONCLUSIONSTwo haplotypes (T-T-b and C-G-a) may be strongly associated with susceptibility to HAPE. Compared with the individual alleles of eNOS gene, the interaction of multiple genetic markers within a haplotype may be a major determinant for the susceptibility to HAPE.

Adolescent ; Adult ; Altitude ; Base Sequence ; Case-Control Studies ; DNA Primers ; Genotype ; Haplotypes ; Humans ; Male ; Middle Aged ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; genetics ; Occupational Diseases ; enzymology ; genetics ; Polymorphism, Genetic ; Pulmonary Edema ; enzymology ; genetics ; Tibet ; Young Adult

Twenty target compounds were synthesized by the reduction reaction of HUANG Minglong and Friedel-Crafts acylation reaction in this study. The inhibitory effects of the new compounds were tested on NO production in LPS-induced mouse macrophage RAW264.7 cells, a cellular inflammation model. The structure-activity relationships were discussed. The structures of target compounds were confirmed by ESI-MS, ¹H NMR and ¹³C NMR. In vitro activity experiments showed that 18 compounds had certain anti-inflammatory effects at the concentration of 40 μmol·L^-1, of which 9a, 8b, 7c and 9c showed strong anti-inflammatory activities, and IC₅₀ of 7c and 9c were comparable to the positive control drug ibuprofen.

Liver fibrosis is a tissue repair compensatory response to liver injury caused by various chronic factors, ultimately leading to liver cirrhosis, liver failure and even hepatocellular carcinoma. Abnormal activation of hepatic stellate cells is the cellular basis of liver fibrosis development. Pepstatin Pr, the derivative of pepstatin A, was isolated from Streptomyces sp. CPCC 202950. Our purpose was to investigate the anti-fibrotic activity of pepstatin Pr and explore its molecular mechanism. Hepatic stellate cell LX-2 was stimulated by TGFβ1 and sub- sequently treated with pepstatin Pr. Its cytotoxicity was detected by sulforhodamine B (SRB) assay. The expression of COL1A1, α-SMA and cathepsin D, signaling proteins TGFβ, Smad and YAP/TAZ were detected by Western blot or real-time PCR. The results showed that pepstatin Pr was not cytotoxic to LX-2 cells. And pepstatin Pr significantly reduced the mRNA and protein expression of COL1A1 and α-SMA, which are important liver fibrosis markers. Pepstatin Pr also repressed the protein expression level of cathepsin D, TGFβ1, YAP/TAZ, the phospholation level of Smad2, and YAP nuclear translocation. In conclusion, pepstatin Pr exhibits anti-fibrotic effects in TGFβ1-stimulaed LX-2 cells by mediating YAP-TGFβ-Smad pathway.