OBJECTIVETo identify the activin A receptor type II-like 1 gene (ACVRL1) mutations in a Chinese family with hereditary hemorrhagic telangiectasia (HHT2).

METHODSThe exons 3, 7 and 8 of ACVRL1 gene of the proband and her five family members were amplified by polymerase chain reaction (PCR), and the PCR products were sequenced.

RESULTSThe proband had obvious telangiectasis of gastric mucosa, and small arteriovenous fistula in the right kidney. All the patients in the HHT2 family had iterative epistaxis or bleeding in other sites, and had telangiectasis of nasal mucosa, tunica mucosa oris and finger tips. ACVRL1 gene analysis confirmed that there is frameshift mutation caused by deletion of G145 in exon 3 in the 4 patients, but the mutation is absent in 2 members without HHT2.

CONCLUSIONThe HHT2 family is caused by a 145delG mutation of ACVRL1 gene, resulting in frameshift and a new stop codon at codon 53.

Activin Receptors, Type II ; genetics ; Exons ; genetics ; Female ; Frameshift Mutation ; genetics ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Telangiectasia, Hereditary Hemorrhagic ; genetics ; pathology

A series of adefovir mono-L-amino acid esters, mono non-steroidal anti-inflammatory drugs carboxylic ester prodrugs with more potent anti-HBV activity and lower nephrotoxicity were designed and synthesized. Adefovir bis (L-amino acid) ester was used as lead compound, according to pathological and pharmacological findings that non-steroidal anti-inflammatory drugs can effectively inhibit the organic anion transporter 1 (hOAT1)-mediated adefovir phosphonic acid pairs of anion transport across tubular basement membrane thereby reducing the nephrotoxicity of adefovir. Flatten design principle was used to introducing non-steroidal anti-inflammatory drugs structural fragments to design and synthesize target adefovir mixture ester prodrugs. HepG2 2.2.15 cell line was used as in vitro anti-HBV activity evaluation model. Five compounds exhibited antiviral activity, and compound 18 showed the most potent anti-HBV activity and relatively high selective index (EC50 3.92 micromol L(-1), SI 9.97). HK-2 cell line was used as in vitro model to evaluate nephrotoxicity. Results suggested the target compounds have lower cytotoxicity than the positive control. Moreover, by analyzing the primary structure and activity relationship of these compounds, it could suggest that mono-L-amino acid ester, mono non-steroidal anti-inflammatory drugs carboxylic ester prodrugs strategy has significant potential in the acyclic nucleoside phosphonates prodrug design.
Adenine ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Amino Acids ; chemical synthesis ; chemistry ; pharmacology ; Anti-Inflammatory Agents, Non-Steroidal ; chemical synthesis ; chemistry ; pharmacology ; Antiviral Agents ; chemical synthesis ; chemistry ; pharmacology ; Carboxylic Acids ; chemistry ; pharmacology ; Cell Survival ; drug effects ; Drug Design ; Hep G2 Cells ; drug effects ; Humans ; Kidney Tubules, Proximal ; cytology ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Molecular Structure ; Organophosphonates ; chemical synthesis ; chemistry ; pharmacology ; Prodrugs ; chemical synthesis ; chemistry ; pharmacology

The aim of this study is to construct a lentiviral vector encoding human growth hormone, and to achieve the long, efficient and stable expression in murine skeletal myoblasts. Primary skeletal myoblasts were isolated from Sprague-Dawley rats and cultured by enzymatic digestion. We tested them by Desmin immunohistochemistry stains and found their viability was up to 94% by Trypan blue. Human growth hormone (hGH) cDNA was subcloned into expression vector pLenti6/V5-D-TOPO to construct recombinant pLenti6/V5-hGH. The pLenti6/V5-hGH and the contructed pLenti6/V5-EGFP were transfected into murine skeletal myoblasts by the Lipofectamin 2000. Through counting by the Confocal Laser Scanning Microscope, we identified the transfection efficency. We added the blasticidin to the 6-well plate with lids and obtained stable myoblasts expressing hGH. The concentration of human growth hormone (hGH) in cell culture medium was detected by Radioimmunoassay (RIA). Polymerase Chain Reaction (PCR) and DNA sequence showed hGH cDNA had been correctly inserted into pLenti6/V5-D-TOPO vector. Bright green fluorescence of the transfected cells could be observed under the Confocal Laser Scanning Microscope after 24 h transfection with pLenti6/V5-EGFP plasmids, and the transfection rate reached 40%. The difference was distinct (P < 0.01) between the pLenti6/V5- hGH groups and control groups in the secretive level of human growth hormone. After 8 weeks, the expression of human growth hormone was still stable. Then, we validated the biological characterization of the rhGH by the enzyme-link immunosorbent assay (ELISA) of the Insulin-like growth factor I (IGF-1). These results demonstrate we have successfully constructed the recombinant pLenti6/V5-hGH plasmids and accomplished rhGH long, efficient and stable expression ectopic in skeletal muscle myoblasts.
Animals ; Animals, Newborn ; Cells, Cultured ; Genetic Vectors ; Human Growth Hormone ; biosynthesis ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Myoblasts, Skeletal ; metabolism ; Plasmids ; genetics ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

Recently, suppressor of cytokine signaling-3 (SOCS3) has been shown to be an inducible endogenous negative regulator of Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway which is relevant in inflammatory response, while its functions in acute liver failure and HBV-induced acute-on-chronic liver failure (HBV-ACLF) have not been fully elucidated. In this study, we explored the role of SOCS3 in the development of mouse hepatitis virus strain 3 (MHV-3)-induced acute liver failure and its expression in liver and peripheral blood mononuclear cells (PBMCs) of patients with HBV-ACLF. Inflammation-related gene expression was detected by real-time PCR, immunohistochemistry and Western blotting. The correlation between SOCS3 level and liver injury was studied. Our results showed that the SOCS3 expression was significantly elevated in both the liver tissue and PBMCs from patients with HBV-ACLF compared to mild chronic hepatitis B (CHB). Moreover, a time course study showed that SOCS3 level was increased remarkably in the liver of BALB/cJ mice at 72 h post-infection. Pro-inflammatory cytokines, interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α, were also increased significantly at 72 h post-infection. There was a close correlation between hepatic SOCS3 level and IL-6, and the severity of liver injury defined by alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, respectively. These data suggested that SOCS3 may play a pivotal role in the pathogenesis of MHV-3-induced acute liver failure and HBV-ACLF.

AIM:To observe the effects of panaxadiol saponins(PDS)on up-regulation of MAPK/ERK signal pathway in bone marrow cells and increase in regulatory T(Treg)cells in spleen tissue of aplastic anemia(AA)mice,and to explore the mechanisms.METHODS:For preparation of immune-mediated AA model,BALB/c mice were exposed to sublethal dose(5.0 Gy)of [60Co]-γradiation, followed by transplantation of lymphocytes from DBA /2 donor mice. BALB/c mice(n=60)were randomly divided into 6 groups,including normal mouse group,AA model group,PDS treat-ment groups at low,medium and high doses,and cyclosporine group as positive control.PDS and cyclosporine were given by gavage for 14 d.The peripheral blood cell counts and bone marrow pathological examination were tested.The protein levels of MEK1/2,p-MEK1/2,ERK1/2 and p-ERK1/2 in the bone marrow cells were analyzed by Western blot and im-munohistochemistry experiment.Flow cytometry was used to detect the proportion of Treg cells in spleen tissue of each group.RESULTS:The peripheral blood cell counts were significantly decreased in AA mouse group as compared with nor -mal mouse group(P<0.05).The bone marrow sections showed markedly inhibition status of hematopoiesis and the de -crease in cellularity.In response to PDS treatment,the peripheral blood cell counts and Treg cells in the spleen tissues of AA mouse treated with PDS were significantly increased in a dose-dependent manner(P<0.05).Treatment with PDS at medium and high doses up-regulated the protein levels of MEK1/2,p-MEK1/2,ERK1/2 and p-ERK1/2 in the bone mar-row of AA mice(P<0.05).CONCLUSION:PDS is effective to enhance recovery of hematopoietic function in AA mice. This effect may be related to up-regulating multiple protein kinases of MAPK/ERK signal pathway in the bone marrow cells of AA mice.In addition,PDS has an impact on immune function of AA mice.

Objective To investigate the effects of IL-21,IL-23 and IL-4 in the pathogenesis of Guillain-Barré syndrome(GBS) of children,and to analyze the influence of intravenous immunoglobulin(IVIG) on the serum levels of IL-21,IL-23 and IL-4 in GBS patients.Methods Forty-one pediatric patients with GBS hospitalized in our department from Jan.2005 to Sep.2012 were studied.According to the time of IVIG administration,patients were divided into group A and group B,given IVIG respectively within the 7th day and the 8th-12th day.According to Hughes score,patients were divided into mild group and severe group.The serum levels of IL-21,IL-23 and IL-4 were detected by means of ELISA before and after treatment.Results Before treatment,the serum levels of IL-21,IL-23 and IL-4 had no signifiant difference between group A and group B.After treatment,the serum levels of IL-21,IL-23 in group A and group B were significantly decreased than those before treatment (all P ＜ 0.05),with no significant difference in IL-4 levels.There was no signifiant difference in group A and group B in the serum levels of IL-21,IL-23 and IL-4 after treatment (all P ＞0.05).In the severe group,the serum levels of IL-21,IL-23 were significantly higher than those in the mild group before and after treatment (all P ＜ 0.05),with no significant difference in IL-4 levels.After treatment,the serum levels of IL-21,IL-23 were significantly decreased than those before treatment in the mild group and the severe group (all P ＜ 0.05),with no significant difference in IL-4 levels.Conclusions The serum levels of IL-21,IL-23 are significantly increased in GBS children,indicating they play an important role in the pathogenesis of GBS.Levels of IL-21 and IL-23 are significantly decreased after administration of IVIG,which showed that IVIG could inhibit the secretion of IL21 and IL-23,reduce cellular inflammatory response,and eventually prevent the development of GBS.IL-4 secretion is not obviously affected with IVIG,it might have a role in the recovery of GBS.

OBJECTIVETo investigate the interaction of deficiency in thrombosis-related gene in a mouse model.

METHODSTo generate mice carrying mutations in alpha-galactosidase A (Gla) and factor V Leiden (Fvl) and analyze the phenotypes, namely, tissue fibrin deposition and thrombus formation in organs.

RESULTSFibrin deposition in organs of mice carrying both mutations in Gla and Fvl was significantly increased compared with that in mice with single mutaton: [Gla(-/0) Fv(Q/Q)+Gla(-/-)Fv(Q/Q)] vs.[Gla(-/0)Fv(+/+)]=(0.28+/-0.03)% vs.(0.07+/-0.007)%, P<0.01; [Gla(-/0)Fv(Q/Q)+Gla(-/-)Fv(Q/Q)] vs.[Gla(+/0)Fv(Q/Q)+Gla(+/+)Fv(Q/Q)]=(0.28+/-0.03)% vs.(0.11+/-0.02)%, P< 0.01. Meanwhile, the number of thrombi on organ sections of mice carrying both mutations in Gla and Fvl was significantly increased compared with the single mutation carrier: [Gla(-/0)Fv(Q/Q)+Gla(-/-)Fv(Q/Q)] vs.[Gla(-/0)Fv(+/+)]=1.9+/-0.7 vs. 0.0+/-0.0, P<0.05; [Gla(-/0)Fv(Q/Q)+Gla(-/-)Fv(Q/Q)] vs. [Gla(+/0)Fv(Q/Q)+Gla(+/+)Fv(Q/Q)]=1.9+/-0.7 vs. 0.3+/-0.1, P<0.05.

CONCLUSIONThese observations demonstrated that there was synergistic effect in Gla and Fvl deficiency in mice. It suggested that there could be a combination of GLA deficiency and FVL or other thrombosis-related gene defect in patients with genetic severe early-onset thrombosis.

Animals ; Factor V ; genetics ; Fibrin ; metabolism ; Immunohistochemistry ; Mice ; Mutation ; Thrombosis ; genetics ; metabolism ; alpha-Galactosidase ; genetics

OBJECTIVETo analyze the effectiveness of Chinese medicine and integrated Chinese and Western medicine for influenza A (H1N1) in the fever clinics and its relevant expenditure.

METHODSA prospective survey on the clinical epidemic observation and follow-up was conducted from July 2009 to October 2009 with a self-developed questionnaire whose contents including the clinical data of the confirmed 149 H1N1 cases and their relevant therapeutic expenditure. The patients were assigned to the Chinese medicine group (22 cases treated by Chinese medicine alone) and integrative medicine group (124 cases treated by both Chinese medicine and Western medicine). The data were processed with descriptive analysis, t test and χ (2), and sum-rank test.

RESULTSThe proportion of clinical recovery of Chinese medicine group (81.8%) was higher than that of integrative medicine group (54.8%) with statistical significance (P=0.02). The average fever durations in both groups were 3.5 to 4 days, showing no significant difference (P=0.86). In the comparisons of average cost of Chinese herbs, drugs, therapies, and total cost, those of the Chinese medicine group were lower than those in the integrative group (P=0.01, P=0.00, P=0.00, P=0.00).

CONCLUSIONSThe H1N1 patients in the fever clinic who received Chinese medicine treatment had a higher clinical recovery proportion than those who received integrated Chinese and Western medicine treatment with lower medical cost. However, due to small sample size of the Chinese medicine group in the study, the conclusion needs further confirmation by studies with large sample size.

Adult ; Costs and Cost Analysis ; Female ; Fever ; economics ; therapy ; virology ; Health Expenditures ; Hospitals ; Humans ; Influenza A Virus, H1N1 Subtype ; physiology ; Influenza, Human ; economics ; therapy ; virology ; Integrative Medicine ; economics ; Male ; Medicine, Chinese Traditional ; economics ; Time Factors ; Treatment Outcome

OBJECTIVETo compare the efficacy and side effects of nedaplatin plus 5-fluorouracil (5-Fu) and cisplatin plus 5-Fu for treatment of stage III-IVa nasopharyngeal carcinoma (NPC).

METHODSA total of 100 patients with NPC proved by histopathology were divided into nedaplatin plus 5-Fu group (NF group) and cisplatin plus 5-Fu group (PF group), 50 cases in each group. NF group: nedaplatin 30 mg/m(2), d1-d3, 5-Fu 500 mg/m(2) d1-d5, repeated every 3 weeks for 2 cycles. PF group: cisplatin 30 mg/m(2) d1-d3, 5-Fu 500 mg/m(2) d1-d5, repeated every 3 weeks for 2 cycles. χ(2) test was used to compare the efficacy and side-effects of the two groups.

RESULTSAll the 100 cases were evaluable and their clinical data in the two groups were comparable. Six patients with complete response were observed, 3 cases in NF group and 3 in PF group. The overall response rates were 86.0% in NF group and 84.0% in PF group, with no significant difference (χ(2) = 0.078, P = 0.779). The rates of leukocytopenia, thrombocytopenia, impairment of hepatic and renal function were similar whereas more patients in the PF group than in the NF group suffered from nausea and vomiting (88.0% vs. 56.0%, P = 0.000).

CONCLUSIONSNedaplatin plus 5-Fu is an effective treatment regimen for NPC. When compared with PF regimen, the response rate is similar. However, NF regimen shows a significant superiority in reducing nausea and vomiting.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Cisplatin ; adverse effects ; therapeutic use ; Female ; Fluorouracil ; administration & dosage ; adverse effects ; therapeutic use ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; drug therapy ; pathology ; Nausea ; chemically induced ; Neoplasm Staging ; Organoplatinum Compounds ; administration & dosage ; adverse effects ; Remission Induction ; Vomiting ; chemically induced

Objective To observe the clinical efficacy of joint needling method combined with ultrasound in the treatment of qi stagnation and blood stasis type of patellofemoral pain syndrome(PFPS).Methods Eighty-six patients with qi stagnation and blood stasis type of PFPS were randomly divided into observation group and control group,with 43 cases in each group.The control group was given western medicine conventional treatment combined with functional exercise,and the observation group was given joint needling method combined with ultrasound treatment on the basis of the control group.Both groups were treated for 2 consecutive weeks.After 2 weeks of treatment,the clinical efficacy of the two groups was evaluated,and the changes in the Visual Analogue Scale(VAS)scores of knee pain and the Kujala scale scores of the two groups were observed before and after treatment.The changes in active range of motion(AROM)of the affected knee joint were compared before and after treatment between the two groups.Results(1)After treatment,the VAS scores of the two groups of patients were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the level of VAS scores,and the difference was statistically significant(P＜0.05).(2)After treatment,the Kujala scores of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the level of Kujala scores,and the difference was statistically significant(P＜0.05).(3)After treatment,the AROM of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the level of AROM,and the difference was statistically significant(P＜0.05).(4)The total effective rate was 95.35%(41/43)in the observation group and 81.40%(35/43)in the control group.The efficacy of the observation group was superior to that of the control group,and the difference was statistically significant(P＜0.05).Conclusion The joint needling method combined with ultrasound can significantly relieve the pain symptoms of patients with PFPS and promote the recovery of knee joint function,and the clinical efficacy is remarkable.