Purpose: To study the clinical treatment and prognosis of gastric cancer in elderly patients. Methods: From 1984 to 1995, 110 old patients with malignant tumor in stomach were treated in this hospital. All 110 patients received surgery with radical resections in 60 cases, palliative resections in 22 cases, laparotomy in 24 cases, and gastrojejunostomy in 4 cases. Results: Postoperative complication was 16. 36%, mortality was 1. 8%. The overall 3- and 5- year survivals for radical resections were 68. 4%, 55. 08%, respectively. Multivariate analyses showed that the favorable factors for prognoses were stage and radical resection. Conclusions: It is recommended that radical resection should be performed for malignant tumors of stomach in old patients. Although the risks of complication morbidity and surgery mortality are more likely to be higher after surgery for the elderly, the outcome would be better if more attention were paid, to improve patients general condition, and correct management after surgery.\;

Purpose:To study the clinical characteristics of primary colorectal non Hodgkin's lymphoma, analyze the prognostic factors, and assess the results of treatment with adjuvant chemotherapy and radiotherapy. Methods:Ninety four patients were restrospectively studied at our institution between 1971 and 1995, and all of them underwent operation and were confirmed primary colorectal non Hodgkin's lymphoma pathologically. Adjuvant chemotherapy and radiotherapy were administered to 77 and 83 patients respectively. The univariate and multivariate analysis were used to estimate the prognostic factors.Results:The 3 , 5 , and 10 year survival rate was 62.5%, 61.0%, and 58.9% respectively. By univariate analysis, the operation type, lymph node metastasis, postoperative chemotherapy, postoperative radiotherapy, clinical stage were the significant prognostic factors for survival ( P

Objective To study the effect of Dachengqi decoction on expression of triggering receptor on myeloid cell 1(TREM-1)in septic rats in order to further provide a theoretical basis concerning the mechanism of this decoction for treatment of sepsis. Methods 100 male Sprague-Dawley(SD)rats were randomly divided into five groups:normal control,sham operation,sepsis model,low-dose and high-dose Dachengqi decoction groups (each,n=20). The sepsis models were reproduced by cecal ligated perforation(CLP). The low-dose and high-dose Dachengqi decoction groups were lavaged separately by low dose(5 mL/kg)and normal dose(10 mL/kg)Dachengqi decoction at 2 hours before CLP and after CLP twice per day(interval 8 hours),and the other three groups were lavaged with 10 mL/kg normal saline. Five rats in each group were killed randomly at the time points of 6,12,24,48 hours after CLP;the abdominal aorta blood and the liver tissue were collected. The plasma levels of TREM-1,interleukin-6 (IL-6),tumor necrosis factor-α(TNF-α)were tested by enzyme linked immunosorbent assay(ELISA). The expression level of TREM-1 mRNA in the liver was measured by reverse transcription - polymerase chain reaction (RT-PCR). Results Compared with normal control group and sham operation group, the plasma levels of TREM-1, IL-6, TNF-α and the expression of liver TREM-1 mRNA were increased significantly in model group. Compared with model group,the above indexes in low-dose and high-dose Dachengqi decoction groups were reduced obviously,the changes being more marked in the high-dose group;the levels of TREM-1,IL-6 at 6 hours after operation and the levels of TNF-αand TREM-1 mRNA at 24 hours after operation in high-dose group were lower than those of low-dose group〔6 hours TREM-1(ng/L):179.19±4.43 vs. 213.86±2.84,6 hours IL-6 (ng/L):136.80±7.70 vs. 162.90±3.87;24 hours TNF-α(ng/L):71.61±5.07 vs. 108.53±6.29,24 hours TREM-1 mRNA:24.33±3.16 vs. 27.22±3.34,all P<0.05〕. Conclusion The partial mechanism of the efficacy of Dachengqi decoction for treatment of sepsis was probably related to the inhibition of TREM-1 expression.

Objective To investigate clinical effect of fluvastatin in the treatment of early diabetic nephropa-thy,to provide a reference for clinical treatment.Methods 90 patients with diabetic nephropathy were selected,the patients were divided into observation group(50 cases)and control group(40 cases).Conventional hypoglycemic ther-apy used in the control group,and valsartan treatment also used.The observation group received fluvastatin on the basis of treatment of control group.The efficacy,urinary albumin excretion rate,inflammatory markers,serum creatinine and other indicators and adverse reactions were compared.Results The effective rate of the observation group was 90.00%,which was significantly higher than 75.00% of the control group,the difference was statistically significant (χ2 =4.325,P <0.05).After treatment,the BUN,UAER,TC,TG,LDL of the observation group were (6.54 ± 1.24)mmol/L,(40.43 ±4.21)μg/min,(3.81 ±0.47)mmol/L,(2.51 ±0.34)mmol/L,(2.41 ±0.64)mmol/L, the improvement was better than the control group,the differences were statistically significant (t =5.547,5.225, 5.457,4.957,5.339,all P <0.05).After treatment,the CRP,IL -6,IL -18,TNF -αin the observation group and control group were significantly improved compared with before treatment,the differences were statistically significant (P <0.05 ).After treatment,the CRP,IL -6,IL -18,TNF -αlevels of the observation group were (4.14 ± 0.87)mg/L,(88.17 ±8.54)pg/mL,(139.64 ±9.48)ng/L,(40.17 ±5.22)ng/L,the improvement was better than the control group,the differences were statistically significant (t =6.914,6.357,5.847,7.054,all P <0.05 ). Conclusion Fluvastatin in the treatment of early diabetic nephropathy has good effect,which will help to improve inflammatory cytokines and proteinuria and protect renal function,it is worthy of clinical application.

Objective:To construct GJB2 gene mutaitons common in Chinese EGFP fusion protein vectors, and to search for better way to study the mechanism of deletion mutaitons in GJB2 gene. Method: Non-fusion protein vectors of 235delC, 299-300 del AT and 176 del 16 bp were first made by point mutaiton methods in vitro. Then expression part of the upper 3 mutations were amplified by PCR and the PCR products were cloned into TA cloning vector. After cutting by restriction enzymes EcoRI/BamHI, three deletion mutaions were inserted into pEG-FP-N1 vector. Sequencing was used to verify the validity of the fusion protein vectors. HEK293 cells were trans-fected with the recombinant DNA samples by the liposome complex method. Results The recombined plasmids were highly expressed in HEK293 cells. Green fluorescence singals were distributed uniformly in cytoplasm. Conclusion; GJB2 mutations common in Chinese EGFP fusion protein vectors were constructed successfully. It may provide a better way to explore the reasons of nonsyndromic hearing loss common in Chinese.

BACKGROUND: Currently, the materials used in clinical practice to repair cruciate ligament of knee joint contain auto-graft bone- mid 1/3 patella tendon-bone (B-T-B), auto-semitendinous muscle, gracilis muscle and allogenic tissue graft. All of them are limited to a certain degree in clinical application. Therefore, people hope to consistently develop artificial ligaments to take the place of auto- and allografts. OBJECTIVE: To investigate the feasibility to construct artificial biological ligament (ABL) by applying a novel biochemical technique using porcine tendon as the raw material. DESIGN: Research of new biological material. SETTING: Department of Orthopedics, Third Affiliated Hospital of Sun Yat-sen University. MATERIALS: Adult pigs of either gender were provided by the Animal Center of Sun Yat-sen University. Scanning electron microscope (SEM, S-520) was provided by Hitachi, Japan, and micro-controlled electron tension-testing device (Model LWK-10B) by Guangzhou Experimental Devices Factory. METHODS: The experiment was performed at the Animal Center of Sun Yat-sen University from January 2004 to June 2005. ABL was established by means of treating porcine tendon with epoxy cross-linking fixation, diversified antigen minimization process, mechanic enhancement modification and surface activating process. Under aseptic condition, a 6-month-old goat's bone marrow was abstracted, and then the bone marrow matrix stem cells were cultured in ABL stent for 3 weeks. Scanning electron microscope was used to observe structure and compatibility of artificial ligament, and mechanics test was used to analyze biomechanics characteristics of ABL. MAIN OUTCOME MEASURES: Structural features, cell compatibility and biomechanics characteristics of ABL.RESULTS: ① Structural features of ABL: The appearance of ABL was similar to that of the normal human ligament. Histological examination showed that the ABL was collagen fibers with no cells. Electron microscope examination revealed that the ABL was composed of hair-looking and fiber-like objects running uniformly in a certain direction and closely parallel-arranged. ② Cell compatibility: Three weeks after xenogenic marrow matrix cells were cultured on the surface of the ABL, it was noted that cells adhered and the matrix secreted by the cells precipitated around the cells. There were no cells found inside the ABL. ③ Mechanical strength of the ligament: The average diameter of ABL was 5 mm and the mechanical test at a speed of 100 mm/min showed that its averaged tensile limit was 927.19 N and the tension-resistant strength was 47.22 N/mm those were close to the corresponding parameters of the normal goat's ACL. The normal goat's ACL was 5 mm. The greatest tensile load was 807.50 N and the tension-resistant strength was 41.13 N/mm.CONCLUSION:As we used the unique biochemical technique and minimized the xenogenic protein immunogenicity of the porcine tendon, ABL has acceptable biomechanical properties and superior biocompatibility. As a substitute of the ligament in the reconstruction of the ACL, ABL has a promising prospect in clinical applications.

Objective:To express the glycoprotein D of herpes simplex virus type 2 (gD2) in the insect cells,and to determine its immunogenicity.Methods:HSV-2 genome was used as the template for amplification of gD2 extracellular domain fragment gene by PCR.The PCR product was inserted into the vector Bacmind,and the constructed recombinant plasmid gD2-Bacmind was transfected into the sf9 cells to package the recombinant baculovirus.The Sf9 cells were infected by recombinant baculovirus seed derived from the forth passage(P4),the titer of P4 recombinant baculovirus was detected by a plaque assay and the expression of recombinant protein gD2 was determined by Western blotting method.The supernatant of infected cells was collected and purified by Ni-NTA affinity chromatography to obtain the target protein gD2,the purified gD2 protein was used to immunize the BALB/c mice in 0, 2, 4 weeks (gD2 group),and PBS was used as negative control(PBS group);the titers of gD2 specific IgG in serum were detected by ELISA assay.Results: The PCR analysis and sequencing results proved that gD2-Bacmind was constructed correctly.The titer of recombinant baculovirus was 2.0×109 pfu·mL-1,the purified gD2 was about 37 000 with expectation,the percentage of gD2 in total protein was 90%.The average value of Log10 of titer of gD2 specific IgG in serum detected by ELISA assay in gD2 group at the sixth week was 4.34,and there was significant difference compared with PBS group(P<0.01).Conclusion: The gD2 expressed by insect-baculovirus expression vector system has the immunogenicity and can be selected as candidate protein for HSV-2 vaccine.

Objective To evaluate effects of tea polyphenols on the mRNA and nucleoprotein expression of Nrf2/Bach1 in human skin fibroblasts (HSFs).Methods Some HSFs were incubated with tea polyphenols at different concentrations of 0,2.5,5,10,20 and 40 mg/L for 24 hours.Methyl thiazolyl tetrazolium (MTT) assay was conducted to evaluate the proliferative activity of HSFs to screen the optimal concentration of tea polyphenols.Then,some other HSFs were treated with tea polyphenols at this optimal concentration for 24 hours.Real-time quantitative PCR (RT-qPCR) was performed to determine mRNA expression of Nrf2 and Bach1,Western blot analysis to measure nuclear expression of Nrf2 and Bach1 proteins,and immunofluorescence assay to determine the distribution of Nrf2 and Bach1 protein in the cell nucleus.Results MTT assay showed that 5 mg/L tea polyphenols had no obvious effects on the proliferation of HSFs,so 5 mg/L was chosen as the optimal concentration of tea polyphenols for subsequent experiments.HSFs cultured without tea polyphenols served as control group.After the treatment,the 5-mg/L tea polyphenol group showed significantly decreased mRNA and nuclear protein expression of Bach 1 (mRNA:0.629 ± 0.077 vs.0.940 ± 0.033,t =6.397,P ＜ 0.05;protein:1.424 ± 0.171 vs.16.966 ± 1.702,t =15.730,P ＜ 0.05),but significantly increased mRNA and nuclear protein expression of Nrf2 (mRNA:1.467 ± 0.076 vs.0.977 ± 0.091,t =7.133,P ＜ 0.05;protein:6.929 ± 0.121 vs.3.537 ± 0.126,t =33.636,P ＜ 0.05) compared with the control group.Immunofluorescence assay showed increased accumulation of Nrf2 protein,but decreased accumulation of Bach1 protein in the nucleus.Conclusion Tea polyphenols can promote the mRNA and nuclear protein expression as well as nuclear distribution of Nrf2,but suppress the mRNA and nuclear protein expression as well as nuclear distribution of Bach 1,finally exerting antioxidative effects.

OBJECTIVETo detect the mutation of PORCN gene in a patient with focal dermal hypoplasia and study the genotype-phenotype correlation.

METHODSPeripheral blood samples were obtained from the family members and control subjects. PCR was carried out to amplify all the exons and adjacent splice sites of PORCN gene and mutation was detected by bidirectional sequencing.

RESULTSA G149C mutation was found at exon 2 of the PORCN gene in the patient, which caused a change from Alanine to Proline at codon 38 (A38P). The patient presented mild clinical manifestations.

CONCLUSIONA new missense mutation (A38P) in the PORCN was detected in the patient, which maybe one of the molecular mechanisms in the pathogenesis of the disease. The relationship between G149C genotype and moderate phenotype might be attributed to the influence of A38P missense mutation towards the corresponding protein, which is different from previous results.

Acyltransferases ; Base Sequence ; Child ; DNA Mutational Analysis ; Female ; Focal Dermal Hypoplasia ; genetics ; pathology ; physiopathology ; Humans ; Membrane Proteins ; genetics ; Mutation ; genetics