Objective To study the effects of hormone,anesthetie and contrast medium on the activities of collagenase lysis.Methods Nuclear tissues were divided into equal amount for different groups and same units of collagenase were used for the lysis.The only difference in the control group from experimental groups was that there were no dexamethasone,lidocaine or omnipaque but existing in experimental groups.Twenty four hours later,the concentrations of the hydroxyproline were determined in different groups and the data were analyzed by statistical software with computer.Results 1.The concentrations of hydroxyproline in the dexamcthasone group,lidocaine group and omnipaque group were significantly lower than that of control group,especially that of lidocaine group,P value was＜0.05 or 0.01.2. The concentrations of hydroxyproline in the dexamethasone+lidocaine group,dexamethasone+omnipaque group,lidocaine+omnipaque group and dexamethasone+lidocaine+omnipaque group were significantly lower than that of control group;and simultaneously lower in dexamethasone group,lidocaine group,omnipaque group respectively;P value was also＜0.05 or 0.01.Conclusion Dexamethasone,lidocaine and omnipaque can individually inhibit the activity of collagenase at different degrees,so they shouldn't be used together with collagenase in treating the lumber disc herniation.

Arthropods of medical importance such as mosquitoes, ticks and sandflies are one of the key drivers of arthropod-borne diseases outbreak, posing a great threat to global public health security. For further understanding the transmission mechanisms of arthropod-borne diseases and establishing the prevention and control measures, a series of experiments of arthropods infection need to be carried out under laboratory conditions. Besides the regular biosafety requirements, some specific considerations need to be taken into account when performing arthropod infection and the infected arthropod rearing. Except for the physical containment composed of biosafety facilities, a comprehensive assessment of the biosafety risks during operations and corresponding preventive measures are also critical to eliminate or mitigate the biosafety risks. In this paper, we introduce our practice in handling mosquito infection with Risk Group 2 pathogens in Arthropod Containment Level-2 (ACL-2) laboratory, with an aim to provide a reference for researchers in related fields.

OBJECTIVETo explore the characteristics of cellular immunity in drug abusers with pulmonary tuberculosis.

METHODSSixty drug abusers with pulmonary tuberculosis and 60 non-drug abusers with pulmonary tuberculosis (control) were enrolled in this study. Three days after establishment of a definite diagnosis, peripheral blood was taken from the patients for lymphocyte subgroup (CD3(+), CD3(+)/CD4(+), CD3(+)/CD8(+) T lymphocyte subgroups and NK cells) examination by flow cytometry, and the CD4(+)/CD8+(+) ratio was calculated. The difference of cellular immunity between the drug abusers and control group was analyzed statistically.

RESULTSCD3(+) and CD3(+)/CD4(+) T lymphocytes subgroups and NK cells of the drug abusers were significantly lower than those of the control patients (P=0.037, 0.028 and 0.015), and the former patients had also significantly lower CD4(+)/CD8(+) ratio (P=0.021). The pulmonary tuberculosis types and CD3(+)/CD8(+) T lymphocyte subgroup were not significant different between the two groups (P=0.053 and 0.85).

CONCLUSIONDrug abuse might depress cellular immunity in patients with pulmonary tuberculosis, which further complicate the treatment of this disease.

Adolescent ; Adult ; CD3 Complex ; metabolism ; CD4 Antigens ; metabolism ; Female ; Humans ; Immunity, Cellular ; Killer Cells, Natural ; immunology ; Male ; Middle Aged ; Substance-Related Disorders ; complications ; immunology ; T-Lymphocytes ; immunology ; metabolism ; Tuberculosis, Pulmonary ; complications ; immunology ; prevention & control ; therapy ; Young Adult

Objective To evaluate the effectiveness of newborn screening of hearing combined with deafness predisposing genes. Methods Through screening,514 newborns who may had the problem of hearing were classified as experimental group and the other 1 028 newborns were classified as control group by MALDI-TOF. Detecting the predisposing genes of GJB2,GJB3,12SrRNA,SLC26A4 including 20 hot spot mutations for these newborns. Results Among 514 subjects, 40 cases were found with deafness gene mutations,and the positive rate was 7. 47%. 7 cases were pathogenic mutation(1 was GJB2 235delC homozygous mutation,6 were GJB3 538C→T heterozygous mutation ),with the rate of 1. 36%,and 33 cases were heterozygous carrier,with the rate of 6. 62%. Among the control group,45 cases were found with deafness gene mutations,and the positive rate was 4. 38%. 3 cases were pathogenic mutation(1 was 12srRNA 1555A→G homozygous mutation,1 was GJB3 538C→T heterozygous mutation,1 was GJB3 547G→A heterozygous mutation),with the rate of 0. 29%,and 42 cases were carriers of heterozygous gene,with the rate of 4. 09%. The positive rate,the pathogenic mutation rate and the heterozygous carry rate of experimental group were higher than that of control group ,and the differences were significant(all p<0. 05). Conclusion The newborns who did not pass the hearing screening should be the target population for test of the deafness predisposing genes. Since the positive rate were still high,if condition permitted,the screening of hearing combined with deafness predisposing genes should be carried out in some areas.

OBJECTIVETo introduce a new method for enlarging the survival area of an expanded flap.

METHODSAfter the skin expander was inflated with enough injection, the first delaying was performed. In the operation, two incisions were made in the skin and subcutaneous tissue superficial to the expander capsule on both sides of the long axis of the expanded flap. After 10 to 14 days, the second delaying followed, in which one pedicle was divided to form a unilateral-pedicled, super-long random flap. When the flap was transferred to the recipient Since 2000,this technique has been used in 16 patients. All the area, the door site was closed directly.

RESULTSflaps survived completely.

CONCLUSIONSThe technique of twice delaying can enlarge the survival area of the expanded skin flap.

Adolescent ; Adult ; Child ; Cicatrix ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; methods ; Surgical Flaps

BACKGROUNDHistone deacetylase inhibitors (HDACIs) have been reported to induce apoptosis in cancer cells. The effects of trichostatin A (TSA) on gastric cancer cells have not been well characterized. This study was aimed to explore the effects and mechanisms of TSA on human gastric cancer SGC-7901 cells.

METHODSThe cells were treated with TSA and analyzed by cell proliferation assay, Western blot, TUNEL assay, flow cytometry by fluorescein isothiocyanate (FITC) conjugated with Annexin V and PI staining, immunofluorescence analysis, analysis of subcellular fractionation, gene chips and real time polymerase chain reaction (PCR).

RESULTSTSA could inhibit cell growth and induced apoptosis in gastric cancer SGC-7901 cells through the regulation of apoptosis-related genes, such as Bcl-2, Bax and survivin. Further study indicated that the pan-caspase inhibitor z-VAD-fmk did not inhibit the apoptosis induced by TSA, and we did not observe the cleavage of poly ADP ribose polymerase (PARP) after TSA treatment too. In addition, apoptosis inducing factor (AIF) and EndoG were found to translocate from mitochondria to nucleus in the immunofluorescence assay and the Western analysis of subcellular fractionation confirmed the result of immunofluorescence assay.

CONCLUSIONSThe apoptosis induced by TSA in gastric cancer SGC-7901 cells involves a caspase-independent pathway.

Apoptosis ; drug effects ; Caspases ; physiology ; Cell Line, Tumor ; Enzyme Inhibitors ; pharmacology ; Gene Expression Profiling ; Histone Deacetylase Inhibitors ; Humans ; Hydroxamic Acids ; pharmacology ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; analysis ; Neoplasm Proteins ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Stomach Neoplasms ; drug therapy ; pathology ; Tumor Suppressor Protein p53 ; analysis ; physiology ; bcl-2-Associated X Protein ; analysis

OBJECTIVETo establish a method for SNP genotyping of multi-genes by allele-specific oligonucleotide probe ligation mediated by a thermostable ligase, and to explore the genetic polymorphisms of drug-metabolizing enzymes in breast cancer patients and their association with chemotherapeutic responses.

METHODS10 SNP loci of enzyme genes related to chemotherapeutic drugs such as taxanes, anthracyclines and cyclophosphamide were selected, and were genotyped for blood samples from 126 breast cancer patients by the established method. Their correlations with therapeutic responses were retrospectively evaluated.

RESULTSThe lower detection limit of genomic DNA by this developed method was 6.25 ng. The fluorescent peak locations of ligation products on ABI PRISM 377 DNA sequencer were accurate and consistent with prospective sizes in bases (Bias range 0.08 - 0.69 bp, x(-) = 0.31 bp, s = 0.18 bp). Same genotyping results were obtained for repeat tests of 8 random samples, which were further confirmed by sequencing analysis. The 10 SNP loci were polymorphic of different frequency in the breast cancer patients. The combinations with GSTP1 genotypes and GSTM1 genotypes were related to anthracycline-based chemotherapy efficacy (P = 0.037), and the low GSTs activity group (GSTP1 variant allele + GSTM1 null) showed the best effects (85.7%). GSTM1 genotypes and their combinations with GSTP1 and/or CYP3A5*3 genotypes were related to taxane-based therapy efficacy (P < 0.05 for all), and both the low GSTs activity group and the drug slow-metabolising group (low GSTs activity group + CYP3A5*3 wild allele) showed better effects (100%).

CONCLUSIONThe established method is reliable and applicable in multiplex SNPs genotyping of multi-genes. SNPs combination may have a better clinical application value for prediction of chemotherapeutic responses.

Adult ; Aged ; Anthracyclines ; therapeutic use ; Antineoplastic Agents ; therapeutic use ; Breast Neoplasms ; drug therapy ; genetics ; Cyclophosphamide ; therapeutic use ; Cytochrome P-450 CYP3A ; genetics ; DNA Mutational Analysis ; methods ; Female ; Gene Frequency ; Genotype ; Glutathione S-Transferase pi ; genetics ; Glutathione Transferase ; genetics ; Humans ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; Retrospective Studies ; Taxoids ; therapeutic use ; Treatment Outcome

OBJECTIVETo study the expression of P57(kip2) and Maspin in the pathological scar and their possible role in the pathogenesis of abnormal scars.

METHODSImmunohistochemistry integrated image analysis and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of P57(kip2) and Maspin in hypertrophic scar, keloid, mature scar and normal skin. Statistics was used to analyze the datas.

RESULTSThe expression of P57(kip2) protein was fixed to fibroblast intranuclear in abnormal scar, and the expression of P57(kip2) protein and P57(kip2) mRNA decreased (P < 0.05). The expression of Maspin protein was fixed to fibroblast cytoplasm and intranuclear in abnormal scar, and the expression of Maspin protein and Maspin mRNA decrease, compared with that in normal group (P < 0.05). There was positive correlation between P57(kip2) protein and Maspin protein expression (P < 0.01).

CONCLUSIONSThe decreased expression of P57(kip2) and Maspin in abnormal scar shows that they are cicatrix-related genes. There is a positive relationship between the two genes. It may be one of the mechanisms of pathogenesis of abnormal scar. It makes effect through fibroblasts.

Cicatrix ; metabolism ; pathology ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p57 ; metabolism ; Fibroblasts ; metabolism ; Humans ; Serpins ; metabolism

OBJECTIVETo summarize the experience of diagnosis and surgical treatment for pulmonary and pleural aspergillosis.

METHODSThe clinical data of cases with pulmonary and pleural aspergillosis were analyzed retrospectively between September 1972 and June 2003. There were 53 cases with pulmonary aspergillosis and 3 cases with pleural aspergillosis. Aspergillus was found preoperatively in 8 patients by sputum culture (5 cases) or needle biopsy of the lung (2 cases) or fibro-bronchoscopic biopsy (1 case). All patients were treated with surgical procedures following X-ray film or CT scan.

RESULTSOf 53 cases with pulmonary aspergillosis, 42 lobectomies, 3 segment-Pneumonectomies, and 8 wedge resections were performed. Of three cases with pleural aspergillosis following eliminating their diseased foci in residual pleural space, two underwent thoracoplasty, one underwent postoperative closed chest drainage for one and an half month with fluconazole injected into residual pleural space repeatedly for 1 month (200 mg/100 ml, 1 time per 2 or 3 days). No operative death and major postoperative complications occurred. None of the patients had recurrent symptoms at follow-up.

CONCLUSIONWe recommend aggressive surgical resection for pulmonary and pleural aspergillosis, and the surgical result is excellent.

Adult ; Aged ; Aspergillosis ; diagnosis ; surgery ; Female ; Humans ; Lung Diseases, Fungal ; diagnosis ; surgery ; Male ; Middle Aged ; Pleurisy ; diagnosis ; surgery ; Pneumonectomy ; methods ; Retrospective Studies ; Thoracoplasty ; Treatment Outcome

OBJECTIVETo investigate the feasibility of life span extension of transformed human embryonic tendon cells (THETC) by reconstitution of the telomerase activity.

METHODSTHETC were transfected by pGRN145 plasmid containing the human telomerase reverse transcriptase (hTERT) cNDA in vitro by molecular cloning technique. The biological characteristics of transfected cells were detected and compared by morphological observation, plate cloning efficiency, soft agar culture, growth curve of cells cultured in different conditions, immunohistochemistry, telomerase activity assay by telomeric repeat amplification protocol (TRAP).

RESULTSThe THETC transfected by pGRN145 plasmid (telT) could express the telomerase activity with extension of life span. The telT maintained the original characteristics of temperature-dependant and serum-dependant, as well as secretion of type I collagen normally and without tendency of malignant transformation.

CONCLUSIONSThe life span of THETC can be prolonged by reconstitution of telomerase activity, which provides the novel experimental methods to establish the standard cells line.

Cell Line ; Cell Survival ; Embryo, Mammalian ; Humans ; Plasmids ; genetics ; RNA-Directed DNA Polymerase ; Telomerase ; genetics ; metabolism ; Tendons ; cytology ; enzymology ; Transfection