Objective To construct eukaryotic cell expression vector of human frangible histone triad (FHIT) gene. Methods A 456 bp cDNA fragment was amplified from the total RNA of normal human thyroid tissue by RT PCR method and cloned into plasmid pcDNA3. The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes Kpn Ⅰ and Bst XⅠ and sequenced by Sanger dideoxy mediated chain termination. The expression of FHIT gene was detected by immunocytochemical methods. Results The results showed that the cDNA fragment included 456 bp entire coding region. The recombinant eukaryotic cell expression vector of pcDNA3 FHIT was constructed, and the sequence of the insert was identical to the published sequence. MM96L cells transfected with the pcDNA3 FHIT plasmid expressed high level of Fhit protein in cytoplasm. Conclusion The recombinant plasmid pcDNA3 FHIT can provide a strong molecular tool for the studies of neoplasm pathogenesis.

Objective To analyze the performance of adsorbing CO2 with zeolite 5A molecular sieve in crew module. Methods Fitting analysis was based on experimental data of adsorbing CO2,O2 and N2 with zeolite 5A molecular sieve and the suitability of isotherm adsorbing equation of Langmuir,Freundlich and BET models to be conducted with Origin software. Then using obtained equations and competitive adsorbing theory,the adsorbed amounts of these three kinds of gas under competitive condition were calculated. Results The constants in equations for adsorbing CO2,O2 and N2 with zeolite 5A molecular sieve were determined,and adsorbed amounts for above three kinds of gases under competitive condition were calculated. Conclusion The adsorbed CO2 amount is affected by high fraction of N2. Therefore zeolite 5A molecular sieve should be modified technically so that its adsorption in N2 might be reduced to an ignored amount.

Objective To observe the clinical effect of mouse nerve growth factor on protrusion of the lumbar intervertebral disc (LDP) treated with operation. Methods 60 cases with LDP received posterior lamina resection nucleus pulposus decompression excise pedicle screws fixation, in which 30 cases (the treatment group) accepted mouse nerve growth factor for 30 d. Their symptoms and signs, visual analogue score (VAS) for pain, and the electromyogram were recorded before and after treatment. Results The VAS in the treatment group improved more than in control group (P<0.05). the incidence of cure and improvement (76.7%) was higher than in the control group (50.0%)(P<0.05). The nerve conduction velocity and action potential of main muscle group also improved more in the treatment group (P<0.05).Conclusion Mouse nerve growth factor can improve the recovery of pain as well as the neural function in patients after operation for LDP.

Objective To observe the effect of the rivaroxaban and low molecular weight heparin calcium to prevent deep vein thrombosis (DVT) after artificial joint replacement. Methods 167 patients included 114 cases for total hip arthroplasty and 53 cases for total knee replacement from Novermber, 2009 to October, 2010 were divided into two groups: observation group (n=84) was treated with rivaroxaban tablets, 6 h after surgery, 10 mg/d, 2 weeks for knee arthroplasty, 5 weeks for total hip arthroplasty; and control group (n=83) was treated with low molecular weight heparin calcium 2500 IU 2 h before and after surgery for 5 d. Ultrasonography was used to detect the deep vein lumen size change and DVT rate of bilateral lower extremities in all patients before and after surgery. Activated partial thromboplastin time (APTT) was tested before surgery, and 2 d, 5 d, 7 d, 14 d, 35 d after surgery. Results ① Incidence of DVT showed that there were 9 cases (10.7%) with DVT in the observation group; there were 15 cases with DVT (18.1%) in the control group. ② Lower limb deep vein lumen size change comparison: there were 23 cases (27.4%) with superficial femoral vein stenosis more than 2 mm in the observation group while 38 cases (45.8%) in the control group, there were 27 cases (32.1%) with popliteal veins stenosis more than 2 mm in the observation group and 47 cases (56.6%) in the control group (P<0.01). ③APTT: APTT prolonged >10 s was 0 in the observation group while 23 cases (27.7%) in the control group (P<0.01). Conclusion Trivaroxaban may prevent DVT after artificial joint replacement and be better than low molecular weight heparins calcium.

Objective To detect the FHIT gene promoter methylation and protein expression in mycosis fungoides(MF).Methods Tissue specimens were collected from 48 patients with MF and 18 normal human controls.FHIT protein expression was determined by immunohistochemistry,and methylation status of FHIT gene by methylation-specific PCR.Results Abnormal methylation of FHIT gene was found in 26(54.2%)out of the 48 specimens.Thirty(63.5%)specimens of MF were negative for FHIT protein,which was observed in all the control specimens.The promoter methylation of FHIT was closely correlated with the protein expression of FHIT,but unrelated to the sex of,tumor staging or lymph node metastasis in patients with MF.Conclusion The FHIT gene promoter methylation may contribute to the inactivation and abnormal expression of FHIT protein in MF.

Objective To investigate the effect of the exogenous fragile hisdidine triad(FHIT) gene on the proliferation and the apoptosis of cutaneous carcinoma cell line A431,and to explore the mechanism of tumor suppression by the FHIT gene.Methods The plasmids pcDNA3-FHIT and pcDNA3-vector were transfected into the cutaneous carcinoma cell line A431 without FHIT gene expression,and then the transfected cells were screened by G418 and the expression of FHIT was determined by the immunocytochemical staining technique.The effect of FHIT on the growth characteristics of cutaneous carcinoma cell line A431 was observed by MTT,colony forming test and flow cytometry.Results Stable FHIT gene expressing A431 cells were produced,the proliferation activity and colony forming capability of A431FHIT were suppressed,whereas the apoptosis was increased.All these differences between A431-FHIT cells and the two control groups of cutaneous carcinoma cells had statistical significance.Conclusion Transfecting the exogenous FHIT gene into cutaneous carcinoma cells line A431can suppress the proliferation of tumor cells,and can also induce apoptosis and cell cycle arrest.

The exprimental teaching of medical immunology is the important constituent.We have explored the existing problems on course content,teaching method and experimental test way according to our practical teaching experience for the past few years.We have also made the preliminary attempt about reforming exprimental teaching of medical immunology.

Objective To investigate the minimal invasive management of cystitis glandularis with invasion of the ureteric meatus.Methods The clinical data of 18 cases were reviewed.Among the 18 cystitis glandularis patients,12 cases were invasion of bilateral ureteric meatus and 6 of unilateral ureteric meatus.Operation or drug treatment was performed on the diseases that can cause cystitis glandularis such as bladder stone,bladder neck stegnosis,external urethral meatus stegnosis and benign prostate hyperplasia.Sensitive antibiotic was administrated in all cases.After placing ureter catheter,transurethral plasma electro-resection was carried out in five patients whose ureteric meatus could be identified.In addition,of thirteen patients with ureteral orifice unable to be identified,there were ten cases with normal renal function,mitomycin was injected under affected membrana mucosa,and then the patient with ureteral orifice identified underwent transurethral plasma electro-resection after placing ureter catheter.On the other hand,the patient whose ureteral orifice still could not be recognized undertaken transurethral electro-resection at first,during which the ureter catheter was put once ureteral orifice had been detected,otherwise,the ureter catheter should be placed through cystoscope if nephritic colic emerged and hydronephrosis aggravated after operation.In those patients with kidney dysfunction,the ureter catheter was put by ureter discission or ureter replantation at first,the transurethral electro-resection could not be executed until the renal function recovered.Following all these procedure above,bladder instillation of drugs regularly,anti-infection and symptomatic treatment were administrated.Results One patient combined with bladder adenocarcinoma received cystectomy,of the other patients,six cases recurred and underwent electrotomy again resulting in no relapse.All nephrohydrops vanished or relieved obviously,nevertheless,urinary tract infection,haematuria and bladder-ureter backstreaming as the cardinal complication developed in some cases.Conclusion In the management of cystitis glandularis encroaching ureteric meatus,total or partial cystectomy can be avoided if ureter draining freely can be ensured,motivation removal,antiinfection,injection of drug under mucosa and preoperative diuresis conduce to the achievement of ureter catheter placing,transurethral plasma electro-resection is still effective methods in treating these cystitis glandularis.

Objective To evaluate effects of ursolic acid (UA) on interleukin?33 (IL?33) expression in HaCaT cells induced by interferon?γ(IFN?γ), and to explore their mechanism. Methods Some HaCaT cells were treated with UA at different concentrations(0, 0.1, 1, 5, 10, 20, 40 and 80μmol/L)for 24, 48 and 72 hours separately. Then, methyl thiazolyl tetrazolium(MTT)assay was conducted to evaluate cell proliferative activity. A cell model of inflammation was established by culture of HaCaT cells with the presence of 200μg/L IFN?γ. Some HaCaT cells were classified into several groups to be treated with IFN?γ(200μg/L)and UA(10 and 15μmol/L)alone or in combination (firstly treated with IFN?γ followed by UA treatment), and those receiving no treatment served as the blank control group. Reverse transcription PCR (RT?PCR) was performed to detect mRNA expressions of IL?6 and IL?33, and Western?blot analysis to measure IL?33 protein expression after 12?hour culture. The expressions of extracelluar signal?regulated kinase 1/2(ERK1/2)and phosphorylated ERK1/2(p?ERK1/2)were also measured by Western?blot analysis after 5?and 60?minute treatments with IFN?γand UA alone or in combination. Results MTT assay showed that the treatments with 5-20μmol/L UA for 24 hours had no effects on cell proliferative activity, while 40-80μmol/L UA could significantly inhibit it at 24, 48 and 72 hours (all P < 0.05). Thus, 10 and 15 μmol/L were chosen as the concentrations of UA for further study. After the treatment with 200μg/L IFN?γ, there was a significant increase in the expressions of IL?33 mRNA(0.812 ± 0.036 vs. 0.412 ± 0.021), IL?6 mRNA(0.947 ± 0.091 vs. 0.595 ± 0.030)and IL?33 protein(1.317 ± 0.119 vs. 0.147 ± 0.036)in HaCaT cells compared with the blank control group(all P<0.05). Compared with the IFN?γgroup, the IFN?γ+10?μmol/L UA group and IFN?γ+15?μmol/L UA group both showed significantly decreased expressions of IL?33 mRNA(0.447 ± 0.042 and 0.438 ± 0.028 respectively, both P<0.05), IL?6 mRNA(0.437 ± 0.099 and 0.350 ± 0.075 respectively, both P<0.05)and IL?33 protein(0.923 ± 0.058 and 0.564 ± 0.113 respectively, both P<0.05). There were no significant differences in IL?33 mRNA expression between the IFN?γ+10?or 15?μmol/L UA group and blank control group(P>0.05), while IL?33 protein expression was significantly lower in the IFN?γ+15?μmol/L UA group than in the IFN?γ+10?μmol/L UA group(P<0.05). The p?ERK1/2 protein expression significantly increased in HaCaT cells treated with IFN?γ for 5 and 60 minutes compared with the blank control group, but significantly decreased in the IFN?γ+15?μmol/L UA group compared with the IFN?γgroup(0.458 ± 0.053 vs. 0.941 ± 0.042 at 5 minutes, 0.302 ± 0.054 vs. 0.509 ± 0.032 at 60 minutes, both P < 0.05). However, no significant differences were observed in the total ERK1/2 protein expression between the IFN?γ+15?μmol/L UA group and IFN?γgroup at 5 or 60 minutes. Conclusion UA can suppress IL?33 expression in HaCaT cells induced by IFN?γ, likely by regulating expressions of the ERK signaling pathway?related proteins.

Objective To study the influence of post-void residual volume on prostate specific antigen (PSA) in patients with benign prostatic hyperplasia (BPH).Methods One hundred and sixty BPH patients who diagnosed by postoperative pathologic manifestations were selected.The patients were divided into 4 groups according to post-void residual volume with 40 cases each:non post-void residual volume (post-void residual volume ＜ 60 ml) group,less post-void residual volume (60 ml≤post-void residual volume ＜ 120 ml) group,middle post-void residual volume (120 ml ≤post-void residual volume ＜ 180 ml) group,mass post-void residual volume (post-void residual volume ≥180 ml) group.The serum PSA levels of 4 groups were observed.Results The PSA level in mass post-void residual volume group [(8.52 ± 6.72) iμ g/L] was significantly higher than that in non post-void residual volume group [(5.26 ± 4.16) μ g/L] and less post-void residual volume group [(5.93 ± 5.20) μ g/L],there were statistical differences (P ＜ 0.05),there was no statistical difference between non post-void residual volume group and less post-void residual volume group (P ＞ 0.05).Conclusions The serum PSA level in BPH patients with non post-void residual volume has no change,while the serum PSA level in BPH patients with mass post-void residual volume is significantly elevated.