OBJECTIVE: To evaluate the effects of the therapy mainly for strengthening healthy energy to promote blood circulation in treating diabetic patients with gangrene. METHODS: A series of 112 diabetic patients with gangrene were investigated before and after treatment in the changes of wounds, clinical symptoms and quality of life. RESULTS: Fifty-eight cases were cured, 12 cases showed response, 36 cases improved, and 6 cases failure of treatment. The total obvious effect rate was 62.5% (70/112) and the amputation rate was 8.9% (10/112). The 104 diabetic cases of three-stage gangrene were classified into grade I, II and III. The total effect rates of the diabetic cases of grade I, II and III were 83.3% (30/36), 55.6% (35/63) and 20% (1/5), and the amputation rates were 0%, 12.7% (8/63), and 40% (2/5) respectively. After the treatment, the clinical symptoms, gangrene number, gangrene area and gangrene depth were obviously improved (P<0.05), and the quality of life was obviously improved (P<0.01), especially in physical condition, emotional well-being and therapeutic effects on patients (P<0.05). CONCLUSION: Strengthening healthy energy to promote blood circulation is effective in treating diabetic patients with gangrene.

OBJECTIVESTo transfer 1.3-fold-overlength genome of HBV expression plasmid into HepG2 cells, and observe the dynamic changes of viral replication as well as expression in the transfected cells.

METHODS4.1 kb fragment of HBV genome, derived from pGEM-HBV, was cloned into Hind III site of the eukaryotic expression vector pCDNA3.1 to construct the recombinant plasmid pHBV. Then HepG2 hepatoma, cells were transfected with pHBV, using Lipofectamine2000 transfection reagent. After 24, 48, 72 hours, the levels of HBsAg and HBeAg in the supernatant of HepG2 cells were determined with Abbott MEIA Kit. Intracellular viral DNA and RNA were analyzed by Southern and northern blot hybridization. In addition, viral-specific proteins (HBsAg and HBcAg) were assayed by immunofluorescence staining.

RESULTSThe expression vector pCDNA3.1 was constructed successfully. The levels of HBsAg were 5.36+-0.25, 13.42+-1.24, 7.52+-0.43, and the values of HBeAg were 9.16+-0.32, 22.75+-1.49, 15.96+-1.03 after 24, 48, 72 hours, respectively. All expected HBV replicative intermediates and specific transcripts were verified by Southern and northern blot analysis. The HBsAg-positive cells peaked after 24 hours, and then dropped slowly. HBsAg positive staining scattered in the cytoplasm, whereas HBcAg lied maily in the cytoplasm apart from nuclears.

CONCLUSIONSThis recombinant plasmid, which initiates viral replication efficiently in infected cells, is expected as a novel tool for investigating HBV replication in vitro.

Carcinoma, Hepatocellular ; pathology ; virology ; DNA, Viral ; genetics ; Eukaryotic Cells ; virology ; Gene Expression Regulation, Viral ; Genome, Viral ; Hepatitis B virus ; genetics ; physiology ; Humans ; Liver Neoplasms ; pathology ; virology ; Plasmids ; genetics ; Transfection ; Tumor Cells, Cultured ; Virus Replication

OBJECTIVETo establish a stable cell line that can replicate hepatitis B virus (HBV) DNA carrying the reverse transcriptase sequence derived from a clinical isolate.

METHODSNested PCR was used to amplify the HBV DNA fragment from the serum. The fragment was cloned into a plasmid that can support HBV replication in vitro by fragment substitution reaction (FSR), followed by the cloning of the neomycin expressing fragment downstream from HBV DNA. G418 selection was conducted after the transfection of HepG2 cells with the recombinant DNA. Real-time PCR and enzyme linked immunosorbent assay (ELISA) were used to screen stable cell lines that can replicate HBV DNA, and the replication of HBV DNA by the cell line was confirmed by using Southern blot analysis.

RESULTSFragment nt55-1654 amplified from the serum DNA was substituted to the plasmid pLL, generating the plasmid p11. The neomycin fragment was cloned into p11, leading to the plasmid p11-neo, and p11-neo was confirmed to be HBV-replication-competent. A stable cell line named 3-10 that can replicate HBV DNA was obtained.

CONCLUSIONSA stable cell line was established that can replicate HBV DNA carrying the reverse transcriptase sequence derived from a clinical isolate. Real-time PCR plus ELISA may help to rapidly screen out stable cell lines replicating HBV DNA.

Cell Line ; Cloning, Molecular ; DNA Replication ; DNA, Viral ; biosynthesis ; Genetic Vectors ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Hepatocytes ; cytology ; virology ; Humans ; Plasmids ; RNA-Directed DNA Polymerase ; genetics ; Virus Replication ; genetics

Aged ; Echinocandins ; administration & dosage ; therapeutic use ; Humans ; Leukemia, Myelomonocytic, Chronic ; pathology ; Lipopeptides ; Male ; Pneumonia, Pneumocystis ; diagnosis ; drug therapy ; pathology ; Trimethoprim, Sulfamethoxazole Drug Combination ; administration & dosage ; therapeutic use ; Uremia ; pathology

Adult ; Aged ; Fatty Liver ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Platelet-Derived Growth Factor ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Receptors, Platelet-Derived Growth Factor ; metabolism ; Receptors, Transforming Growth Factor beta ; metabolism ; Young Adult

A new computer-assisted vector-cardiogram analyzing system Model TJ-IV developed based on Model TJ-III, has been using in the routine clinical work in order to evaluate its features and performances. The system employs a 586 computer with a CPU of 120 MHz, a special low-noise amplifier, a 12 bit A/D tranducer and the C language for programming. The examinations of 206 cases were performed and all the vector-cardiograms were analyzed by the computer system and by manipulative methods respectively. In comparison with the manipulative methods the system has a very high accuracy of picture-recognition. The accuracy for distinguishing the onsets and terminals of orthogonal ECG waves is 98% while that for distinguishing the peaks and troughs of the waves is 100%. These waves include P, Q, R, S, R' and S' waves. The new system is capable to provide the parameters of more than 591 items, including 46 newly-developed diagnostic parameters. The testing and analyzing of 12 parameters of orthogonal ECG and plane VCG have proved that the results of the aboved two methods have no difference. The new system has a very high accuracy of picture-recognition and index calculation with many technical problems existing in the old versions, solved--a great improvement of safety and anti-interference and an increase of the detecting & diagnostic speed.
Adolescent ; Adult ; Aged ; Automatic Data Processing ; Computer Systems ; Diagnosis, Computer-Assisted ; instrumentation ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnosis ; Reference Values ; Reproducibility of Results ; Signal Processing, Computer-Assisted ; Software ; Vectorcardiography ; instrumentation