Objective To investigate the awareness of osteoporosis and to evaluate the effect of related health education among Beijing community residents.Methods Total 1 000 patients,who visited in 4 community health service centers in Beijing from June to December 2011,were selected as study participants.The questionnaire survey on the awareness of osteoporosis was conducted in all participants,and the items of questionnaire included the risk factors,symptoms,prevalence of osteoporosis,the recommended daily calcium intake and risk of falls.Five hundred participants in 2 health service centers were designated as intervention group and 500 in another health service centers as control group.In intervention group the health education of osteoporosis was conducted every 2 to 3 months and the outpatient follow-up was provided every 2 months;the contents of health education included the clinical manifestation,prevention and control of osteoporosis,and the preveution of fall.In control group only the general outpatient consultation and referral were provided.The second questionnaire survey was conducted 2 years after intervention,the results were compared with baseline and the intervention effect was evaluated.Results Before intervention,58.0％ (580/1 000) participants had exercise,43.1％ (431/1 000) participants took calcium supplement and vitamin D,and 12.1％ (121/1 000)took medication following doctor's advance.The rates of fall prevention and administration of calcium vitamin D in intervention group before intervention [27.8％ (139/500) and 38.7％ (194/500)] were lower than those in control group [33.8％ (308/500) and 47.5％ (237/500)].After two-years of intervention the adoption rate of health behavior in intervention group was improved markedly,the highest one was diet control [increased by 38.4％ (192/500)] and the lowest one was medication [increased by 17.3％ (87/500)].Compared to control group the diet control rate in intervention group was significantly higher [74.0％ (370/500) vs.23.6％ (108/457),x2 =261.887,P=0.000].In addition,unhealthy behaviors were also reduced:55.0％ participants reduced caffeine consumption and 72.2％ participants reduced sodium intake in intervention group.Conclusion The awareness of osteoporosis among Beijing community residents is not satisfactory,but intervention focusing on osteoporosis prevention education is effective to change their behavior.

Objective To evaluate the safety of surgical procedures for patients after heart valve prosthesis implantation.Methods Clinical data of 12 cases with heart valve prosthesis implantation undergone other surgical treatment from November 1996 to December 2005 were retrospectively analyzed.All the cases had routine oral warfarin with prothrombin time (PT) of 20.0—28.3 s averaged 23.5 s, international normalized ratio (INR) for prothrombin of 1.79—2.23 averaged 1.95 and heart functional class Ⅰ—Ⅲ.Among them,appendectomy was performed in three cases with acute appendicitis,reposition and repair in one with inguinal hernia,radical gastrectomy in two with gastric carcinoma,left hemicolectomy in one,cholecystectomy in three,left femoral head replacement in one,and bilateral high ligation and ablation of great saphenous vein in one.Elective surgical operation was performed in seven cases,and emergency operation in five.In those with elective surgery,warfarin was stopped 2—3 days before operation,while 5—10 mg vitamin K_1 was injected intramuscularly 6—8 hours before emergency surgery with preoperative median PT of 15.1 and 15.3 s and median INR of 1.24 and 1.30,respectively.In operation,5—10 mg vitamin K_1 were injected intravenously into the patients by drip depending on their bleeding on the surface of wound.ECG,blood pressure,hemoglobin and oxygen saturation were routinely monitored for all the cases intraoperatively and postoperatively.For the cases with heart function above class Ⅱ,fluid infusion was adjusted based on intubated central venous pressure,and for those with general anesthesia,analyses of blood gases and electrolyte were monitored routinely in operation.Results OPeration time averaged 20—160 rain in all the 12 patients,with blood loss 5—280 ml in average and without complications of massive hemorrhage,thrombosis and heart failure.Conclusions Surgical operation was safe for patients with heart valve prosthesis implantation,if preoperative PT and INR were adjusted to about 15 s and 1.30,respectively by cessation of warfarin or application of vitamin K_1,combined with careful manipulation and strengthened perioperative management.

OBJECTIVETo evaluate the clinical and laboratory features of pediatric inv(16) acute myeloid leukemia (AML) retrospectively.

METHODDual color fluorescence in situ hybridization (D-FISH) using a LSI CBFβ inv(16) break apart probe labeled by Spectrum red and Spectrum green was performed in 15 acute myeloid leukemia cases, including 13 cases with or without abnormal eosinophils but with positive core binding factor β (CBFβ)-MYH11 fusion transcript detected by RT-PCR, and 2 cases with trisomy 8 (+8). The results were compared with the morphology, immunophenotype, karyotype and RT-PCR.

RESULTMorphologically, 12 cases were diagnosed as M(4)EO, 2 as M(4), and 1 as M(2a). Immunophenotypically, all 13 AML cases with inv(16) showed positive expression of CD(13) and CD(33), but without the expression of any lymphoid lineage antigens. Karyotyping analysis with G-banding detected inv(16) in 10 AML cases, including 9 M(4)EO cases and 1 M(2a), but only 5 positive cases were detected using R-banding technique. Among them, 2 cases had simultaneous +8 and trisomy22 (+22), one had +22 only in addition to inv(16). D-FISH revealed a CBFβ-MYH11 rearrangement in 13 cases of AML with positive RT-PCR results, and the mean positive rate of cell detection was 55.15% (range 37.0% - 86.0%). The complete remission rate (CR) and median survival period in this series of inv(16) AML were 81.5%and 11 months, respectively, of whom, 8 cases were still in CR. Relapse and karyotypic evolution were seen in case 5 with +8, +22 in addition to inv(16).

CONCLUSIONAML with inv(16) is a special subtype. Most cases belong to M(4)EO. Its prognosis is good in general, but it seems to be an unfavorable feature for AML with inv(16) and +8, +22 simultaneously, especially with karyotypic evolution. For detection of inv(16), G-banding technique is evidently superior to R-banding technique. D-FISH combined with RT-PCR are more sensitive and reliable than chromosome banding analysis.

Adolescent ; Child ; Child, Preschool ; Chromosome Deletion ; Chromosome Inversion ; Chromosomes, Human, Pair 16 ; genetics ; Eosinophilia ; pathology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Male ; Prognosis ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction

0.05)in the former part of the surgery that was before the beginning of the surgery via anterior route.But in group B,only propofol for sedation was used for the patients during the latter part of the surgery via the anterior route or while the nerve plexus was blocked; during this time in group A,the addition of fentanyl and vecuronium were still intermittently necessary to maintain the general anesthesia.The duration between the completion of surgery and the recovery of spontaneous breathing,times for initial conscious reaction such as opening the eyes following an order, extubation and from extubation to complete recovery were significantly shorter in group B than those in group A(all P

OBJECTIVETo explore clinical and experimental features of 28 cases of childhood acute myeloid leukemia (AML) with 11q23/MLL gene rearrangements.

METHODSKaryotypes of 234 cases of de novo childhood AML were analyzed using short-term culture of bone marrow cells and R-banding. The fusion transcripts involving MLL gene and partial tandem duplication of MLL (MLL-PTD) were detected by multiple reverse transcription polymerase chain reaction (RT-PCR) assay. Two cases with 11q23 translocation by karyotypic analysis but with negative result of multiple RT-PCR were studied with MLL-dual-color fluorescence in situ hybridization (D-FISH).

RESULTSR-banding karyotypic analysis has revealed 20 cases with 11q23 translocation (14 cases with M5, 4 cases with M4, 2 cases with M2), including 12 cases with t(9;11)(p22;q23), 3 cases with t(1;11)(q21;q23), 2 cases with t(6;11)(q27;q23), 1 case with t(11;19)(q23;p13), 1 with t(5;11)(q31;q23), and 1 with t(X;11)(q24;q23). Eighteen cases with 11q23 translocation having fusion transcripts involving MLL genes were confirmed with multiple RT-PCR; 2 cases showed negative results, but they were confirmed to have MLL rearrangements by D-FISH. MLL-PTD was also detected in 8 cases (4 cases M5, 2 cases M4, M2 and M6, one case each) from the other childhood AML cases. The total incidence of 11q23/MLL gene rearrangements was 11.97% (28/234), and most of patients(85.7%, 24/28) were M4/M5. The complete remission (CR) rate after treatment for the 28 cases with MLL rearrangements was 53.8%, the difference was significant by statistics (P< 0.05) compared with 90.5% for the control group (M4/M5 childhood AML with other karyotypic abnormalities or normal karyotype). Of them, 2 cases receiving intensive chemotherapy survived for 81 and 66 months, respectively, 4 cases receiving allogeneic stem cell transplantation survived for 21, 20, 16 and 11 months, respectively, and are still alive with CR. The medium survival (MS) time for 28 cases with 11q23/MLL rearrangements was 11 months, whereas the MS for control group was 15 months. The difference was not statistically significant(P> 0.05).

CONCLUSIONThe 11q23/MLL rearrangements is highly correlated with the occurrence of monocytic leukemia (M4 and M5). The 11q23 translocation and MLL-PTD are mutually exclusive, though both are indicative of poor prognosis. Intensive chemotherapy and allogeneic stem cell transplantation may ameliorate the clinical outcome. Multiple RT-PCR combined with karyotypic analysis and D-FISH are useful for screening the 11q23/MLL rearrangements in childhood AML.

Adolescent ; Child ; Child, Preschool ; Chromosomes, Human, Pair 11 ; Female ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Infant ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; drug therapy ; genetics ; mortality ; Male ; Myeloid-Lymphoid Leukemia Protein ; genetics ; Remission Induction ; Translocation, Genetic ; Treatment Outcome

OBJECTIVETo collect background information on drug-resistant HIV-1 strains in various regions before the start of nation-wide antiretroviral therapy in China.

METHODSTwenty percent of the 2,000 blood samples from antiretroviral therapy naive patients collected for the 2nd national HIV molecular epidemiology survey (NHMES) in 2002 were randomly sampled for this study. The entire protease gene and 20-230 amino acids of the reverse transcriptase gene were amplified by PCR from provirus DNA and sequenced. The results were analyzed with HIV db-Drug Resistance Algorithm and genotypic resistance mutations were determined to particular anti-HIV drugs.

RESULTSTotally 164 protease gene sequences and 138 reverse transcriptase gene sequences were obtained from patients; 0.61% of 164 sequences displayed primary resistance mutations in the protease gene, whereas 99.39% carried 1 or more secondary mutations. Genotypic resistance to at least one nucleoside reverse transcriptase inhibitors (NRTI) was present in 5.80%,and resistance to at least one non-nucleo side reverse transcriptase inhibitors (NNRTI) was present in 1.45% of samples.

CONCLUSIONThe prevalence of genotypic drug resistance is very low in drug-naive HIV infected patients from 21 provinces of China tested in this study. Laboratories participated in the NHMES have organized a network to provide drug resistance monitoring service in the current nation-wide antiviral treatment program in China.

Anti-HIV Agents ; therapeutic use ; China ; epidemiology ; Drug Resistance, Viral ; Genotype ; HIV Infections ; drug therapy ; epidemiology ; virology ; HIV Protease ; genetics ; HIV Protease Inhibitors ; therapeutic use ; HIV Reverse Transcriptase ; genetics ; HIV-1 ; genetics ; Humans ; Mutation ; Reverse Transcriptase Inhibitors ; therapeutic use ; Sentinel Surveillance

OBJECTIVETo find a new method of treating hepatocellular carcinoma with melittin by way of using the melittin gene.

METHODSThe recombinant adenoviruses carrying the melittin gene and alpha-fetoprotein (AFP) promoter (Ad-rAFP-Mel) were constructed through a bacterial homologous recombinant system. The efficiency of the adenovirus mediated gene transfer and the inhibition effect of Ad-rAFP-Mel on the proliferation of hepatocarcinoma cells were determined by X-gal staining and MTT assay respectively. The tumorigenicity of hepatocarcinoma cells transfected by Ad-rAFP-Mel and the antitumor effect of Ad-rAFP-Mel on the transplanted tumors in nude mice were detected in vivo.

RESULTSThe mRNA of the melittin gene was transcripted in HepG2 hepatocellular carcinoma cells transducted by Ad-rAFP-Mel. The efficiency of adenovirus mediated gene transfered to BEL-7402 hepatocarcinoma cells was 100% when the multiplicities of infection (MOI) of Ad-rAFP-Mel was 10 in vitro and was high in vivo as well. The inhibitive rates of Ad-rAFP-Mel and Ad-rAFP for BEL7402 cells were 66.2%+/-2.7% and 2.9%+/-2.3% (t = 30.83) by MTT assay. The inhibitive rates of Ad-CMV-Mel for BEL7402, SMMC7721 and L02 cells were 58.9%+/-9.6%, 65.9%+/-3.8%, 31.7%+/-1.2%, respectively, and those of the Ad-rAFP-Mel were 6.2%+/-2.7%, 16.1%+/-6.6%, 7.5%+/-3.3%, respectively (t = 1.27; t = 11.31, and t = 12.12, vs. Ad-CMV-Mel group in same cells). The tumorigenicity rates of hepatocarcinoma cells transfected by Ad-rAFP-Mel were decreased. A significant antineoplastic effect was detectd on transplanted tumor in nude mice by intratumoral injection of Ad-rAFP-Mel.

CONCLUSIONAd-rAFP-Mel can inhibit specifically the proliferation of AFP-producing human hepatocarcinoma cells in vitro and in vivo. It suggests that animal toxin gene can be used as an interesting antitumor gene.

Adenoviridae ; genetics ; Animals ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Liver Neoplasms, Experimental ; pathology ; Male ; Melitten ; biosynthesis ; genetics ; pharmacology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transcription, Genetic ; drug effects ; alpha-Fetoproteins ; biosynthesis ; genetics

OBJECTIVETo characterize the human immunodeficiency virus (HIV) -specific T lymphocyte responses and identify the immunodominant regions in Chinese HIV-1 recombinant subtype B/C chronic infectors at complete genome level.

METHODSTwenty-five HIV-1B/C recombinant chronic infectors were screened for their specific T lymphocyte responses to a panel of peptides corresponding to the complete HIV-1 subtype B genome by gamma interferon ELISPOT assay. Kruskal-Wallis nonparametric analysis of variance was used to test significant differences across gene regions, and Tukey pairwise analysis was used to identify differences between gene regions. Spearman rank correlation was used to assess the relation between responses. Results The order of recognized frequencies of specific T lymphocyte responses to HIV proteins was Nef>Vpr>Gag>Pol>Vpu>Env>Rev>Vif>Tat. When adjusted for protein length, Nef, Vpr, Gag, and Pol were the most intensely targeted proteins and the central region of Nef, Gag p24, Pol RT, and Vpr was most frequently recognized. No significant correlation was observed between the magnitude of IFN-gamma production of HIV-l-specific T lymphocyte responses and plasma viremia, breadth of response and CD4 counts. Conclusion The central region of Nef, Gag p24, Pol RT, and Vpr is most frequently targeted in HIV-1 B/C recombinants chronic infectors. HIV-l-specific T lymphocyte responses and plasma viremia or CD4 counts play no protective role at complete genome level in these infectors.

Adolescent ; Adult ; Asian Continental Ancestry Group ; CD4 Lymphocyte Count ; Chronic Disease ; Female ; HIV Infections ; immunology ; HIV-1 ; immunology ; Human Immunodeficiency Virus Proteins ; Humans ; Male ; T-Lymphocytes ; physiology ; Viral Load ; Young Adult

OBJECTIVETo observe the induced apoptosis of recombinant adenovirus carrying melittin gene (Ad-rAFP-Mel) for hepatocellular carcinoma cell line (BEL-7402).

METHODSThe morphological observe, DNA electrophoresis, TUNEL and Flow cytometry assay were used to study the apoptosis of BEL-7042 cell line transfected by Ad-rAFP-Mel.

RESULTSThe morphological changes and apoptosis of BEL-7402 transfected by Ad-rAFP-Mel were confirmed with microscopy and DNA electrophoresis, TUNEL, Flow cytometry assay. The DNA ladder could be demonstrated on DNA electrophoresis in Ad-rAFP-Mel group. The apoptosis rates of BEL-7402 cells in Ad-rAFP-Mel, Ad-rAFP, and control groups were (21.5+/-2.4)%, (10.5+/-4.4)% and (3.0+/-1.4)% respectively by TUNEL assay (F = 38.0, P < 0.05) and were (7.3+/-0.5)%, (3.9+/-0.1)% and (0.8+/-0.1)% respectively by flow cytometry assay (F = 415.1, P < 0.05).

CONCLUSIONIt seems that melittin inducing apoptosis might be one of the antitumor mechanisms.

Adenoviridae ; genetics ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Gene Expression ; drug effects ; Gene Silencing ; drug effects ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Liver Neoplasms ; pathology ; Melitten ; biosynthesis ; genetics ; pharmacology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transcription, Genetic ; drug effects ; Transfection

In a 5L fermentor the production conditions of alkaline protease gene engineering strain BA071 were investigated. The maximum activity of alkaline protease reached 24,480 u/mL in 40 hours of fermentation by combination of enhancing aeration and changing the agitation rate. The fast purification method of recombinant protease was conducted with FPLC (Fast Protein Liquid Chromatography). The crude enzyme, treated with ammonium sulfate fractionation and decolored with DEAE-A-50 and polyethylene glycol concentration, was purified with CM-Sephadex-C-50 and Sephadex-G-75. The purified enzyme appears homologous on SDS-PAGE. The purity of enzyme was increased 76.2 times. SDS-PAGE analysis showed that the molecular weights of expressed recombinant products were about 28 kD. The optimal reaction pH and temperature of recombinant enzyme were at pH11 and 60 degrees C, respectively. The recombinant enzyme exhibited high temperature tolerance and was stable at a wide range of pH. Ca2+, MG2+ can enhance the stability of the recombinant enzyme. While the protease activity of the enzyme was strongly inhibited by Hg2+, Ag+, PMFS [symbol: see text] DFP, and was not affected by SDS and Urea.
Bacillus ; genetics ; metabolism ; Enzyme Stability ; Fermentation ; Genetic Engineering ; Metals ; pharmacology ; Recombinant Proteins ; biosynthesis ; Serine Endopeptidases ; genetics ; isolation & purification ; metabolism