Spreaded and immobilized the bacterial cells entraped in PVA on pieces of cotton cloth as porous carrier. The conditions for preparing immobilized cells were as below: cell concentration, 20 mg wet weight/ml; PVA concentration, 5%; spreading amount, 0.3ml/cm~2; immobilization for 12 hours in saturated boron acid solution. Then activated the immobilized cells in buffer containing dyes. Thus, immobilized cells with high decolorizing activity were obtained.In columns packed with immobilized cells, the decolorization efficiencies of continuous influent and intermissional influent were compared with each other. In twenty days, the decolorization rates were both higher than 90%; then the decolorization rate of continuous influent decreased to about 60% after 60 days while it still reached 80% in case of intermissional influent. The efficiency of the later was distinct higher than that of the former.

Cytotoxic T lymphowcytes (CTL) play a major role in host anti-tumor immune responses. A major obstacle to the application of adoptive immunotherapy in the treatment of human maligancy is the inability to generate enough activated CTLs since the cytotoxic T cell undergoes activation induced apoptosis during destroying tumor cells. It is important to study how to limit activaton induced apoptosis of T cell so as to maximize the number of CTL and to enhance the tumor cytotoxicity. We have used CD3-induced Jurkat cell line as an activated T cell apoptosis model and introduced the anti-sense ICE cDNA into Jurkat T cells with retroviral vector. The effect on apoptosis of Jurkat cell induced by anti-CD3 or anti-Fas monoclonal antibody was evaluated after transfection with antisense human ICE. We found that the level of ICE expression in Jurkat cell transduced by the vector decreased and apoptosis was minimized in antisense ICE-transfected Jurkat cell after anti-CD3 or anti-Fas treatment. These results suggest that antisense blocking of ICE expression can partially inhibit Jurkat cell apoptosis induced by anti-CD3 or anti-Fas. Thus, applying antisense blocking of ICE to gene therapy may block the apoptosis of activated T cells, furthennore, enhance the antitumor effect.

P53 gene (exon7～8) mutatins and p53 proteins and HPV 6，11，16，18-DNA were examined in 49 cervical carcinoma by immunohistochemistry, polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) in order to investigate their role and mutual relation and clinical significance in the onco genesis of cervical carcinoma. The results showed that first, p53 proteins posit ive rate was 48.98％, and not outstandingly related to the differentiation and the invasive degree of cervical carcinoma(P＞0.05); the defects of P53 gene (exon7～8) were not found but P53 (exon7～8) mutations were detected in 7 of 49(14.29％) cervical carcinoma; then, HPV16-DNA positive rate was much higher than HPV6,11,18-DNA positive rate respectively(P＜0.001),and the different HPV-DNA was simultaneously tested in one cerv ical carcinoma; last, not all cases of P53 mutations had p53 proteins posi tive, but the cases of P53 mutations and p53 proteins negative certainly had HPV infections, and HPV positive cases were much more than its negative one in the cases of p53 proteins positive(P＜0.001). These results proved that the oncogenesis of cervical carcinoma is mainly associated with HPV16 infections, and second related to P53 (exon7～8) mutations. p53 proteins positive results from P53 mutations or/and HPV infections in cervical carc inoma.

Objective: To investigate the cytotoxicity of ICE gene transfection in Combination with Antitumor Chemicals killing Hepatocellular Carcinoma Cells in vitro.Methods: The recombinant plasmid pLXSN-hICE was transferred to virus packing cell PA317 by electroporation method. And then the retrovirus containing human ICE cDNA generated by these PA317 cells were used to transfect human hepatocellular carcinoma cell line HepG2. The apoptosis of transferred cells were examined by gel electrophoresis. The influence of chemotherapeutic drug Carbo-platin to the proliferation of hepatocellular carcinoma cell line SMMC7721 and its derivative cells(SMMC7721-hICE,SMMC7721-antisence hICE,SMMC7721-neo)was observed by incorporation of 3H-TDR. Results: Electrophoresis of DNA displayed the apoptosis ladder of HepG2 transfected by ICE gene. The proliferation of SMMC7721-hICE was significantly suppressed in vitro induced by Carbo-alatin compared to the other three cell lines. Conclusion: ICE gene transfection could greatly increase the susceptibility of SMMC7721 cells to apoptotic cell death following chemotherapy. These findings suggest that combining ICE gene transfection with utilizing antitumor drugs would represent a novel approach for the effective treatment of hepatocellular carcinoma.

To observe the effect of Helicobacter pylori(H.priori) on cell gap junction ultrastructure of gastric epithelial cells,and to explore carcinogenic mechanism of H.priori from the changes of cell gap junction,BGC-823 cells were co-cultured with different H.prlori strains for 24 h and 48 h.The cell gap junction ultrastructure was observed under transmission electron microscope with sample preparation of fixation and embedding in situ.In 70 patients with gastric eancer(GC),H.priori was detected by rapid urease test,basic fuchsin stain and 14C-urea breath test.The CagA gene of H.prlori was determined by PCR and the cell gap junction ultrastructure was observed under transmission electron microscope.More cell gap junctions and junction complexes of BGC-823 cells were found in control group without H.priori.Groups with H.priori had less number of cell gap junctions,less number of junctions/unit perimeter,shorter length of junctions/unit perimeter,and bigger width of the intercellular space,comparing to control groups without H.priori(P＜ 0.001 or P＜ 0.005).The number of cell junctions and the number of junctions/unit perimeter in the groups co-cultured with NCTC J99,GC 01 and NCTC 11639(CagA+) were less than that in the groups co-cultured with NCTC 12908(CagA-)(P ＜0.001 or P＜0.05),and the length of junctions/unit perimeter in the groups co-cultured with NCTC J99 and GC 01 was shorter than that in the groups co-cultured with NCTC 12908 (P＜ 0.001).In patients with GC,the number of cell junctions,the number of junctions/unit perimeter and the length of junctions/unit perimeter in group H.priori infection were all less than those in group without H.prior/infection(P ＜0.001),and that in CagA+ H.prlori group were less than that in CagA- H.prlori group,but its smallest width of the intercellular space was longer than that in CagA- H.prlori group.The above results showed that the changes of cell gap junction of gastric epithelial cells were associated with H.prlori infection especially CagA+ H.prlori infection.

Objective:To study the effect of metformin on proliferation,cell cycle and apoptosis of U937 cells.Methods: U937 cells were treated with different concentrations of metformin,collected cells in 24,48 and 72 hours.Subsequently,cell proliferation was assessed by CCK-8 assay,and the cell cycle and apoptosis were analyzed by flow cytometry (FCM).The expression of Bcl-2,Bax,p-AMPK,p53 were determined by Western blot.Results: The proliferation of U937 cells was inhibited by metformin in a time-and dose-dependent manner.Metformin-treated cells were arrested at G0/G1 phase,the cell frequency at G0/G1 phase was increased in a time-and dose-dependent manner.Metformin also induced cell apoptosis in a dose-dependent manner.It showed that 20 mmol/L metformin induced cell apoptosis in a time-dependent manner.The expression of p-AMPK,p53,Bax was up-regulated while Bcl-2 expression was down-regulated after metformin treatment.Conclusion: Metformin could inhibit the U937 cell proliferation,block the cell cycle at G0/G1 phase,and induce cell apoptosis,which may partially be attribute to the up-regulation of Bax,down-regulation of Bcl-2,activation of AMPK/p53 signaling.

Objective To explore the influence of continuing nursing care on sexual quality of patients after laparoscopic subtotal hysterectomy.Methods Seventy-eight patients with selective laparoscopic subtotal hysterectomy enrolled from March 2014 to April 2015 were numbered according to their enrollment date,and divided into control group and intervention group randomly.Both groups were conducted laparoscopic subtotal hysterectomy under general anesthesia.After the surgery,the control group was treated with standard nursing care and the intervention group received continuing nursing care beyond the standard nursing care.Participants' sexual quality of 3 months and 6 months after surgery were evaluated using the female sexual function index (FSFI) Results The scores of FSFI of intervention group were significantly higher than those of the control group.Conclusion The continuing nursing care of patients after laparoscopic subtotal hysterectomy can effectively improve patient's short-term sexual quality,which is worth of being applied in gynecology nursing care in the future.

Objective To provide an objective basis for differential diagnosis of pancreatic diseases through quantitative analysis of the different features of contrast-enhanced endoscopic ultrasonography (CE-EUS). Methods A total of 32 patients with suspected or confirmed pancreatic neoplasms or chronic pancreatitis and 19 patients who underwent EUS due to other digestive problems other than pancreatic disease were enrolled. Features of blood perfusion of the regions of interest during CE-EUS were analyzed quantitatively. The findings were compared with cytological and/or histopathological results of EUS-FNA and/or surgery.Results Quantitative analysis of CE-EUS showed peak intensity (PI) value of 19 normal pancreas was 0.648 ±0. 174, which was statistically different from that of pancreatic cancer and pancreatic cystic lesions. Based on ROC, the cutoff of differential diagnosis was 0. 505, and the sensitivity and specificity were 100. 0% and 84. 2%, respectively. PI value of 6 chronic pancreatitis was the highest (0. 772 ±0. 106). In pancreatic neoplams, PI values of pancreatic carcinoma, pancreatic cyst and pancreatic endocrine tumors were significantly different. Based on a cutoff of 0. 195, the sensitivity and specificity of differentiation of pancreatic carcinoma and pancreatic cyst were 85.7% and 87.5%, respectively. PI value of 14 pancreatic carcinoma and that of 4 pancreatic endocrine tumors were 0. 321 ± 0. 119 and 0. 763 ± 0. 115, respectively. Through the comparison between the AT and TTP of the focal lesions and surrounding pancreatic parenchyma, 78.6% pancreatic carcinoma showed slow falling-in and rapid wash-out and all the endocrine tumors showed rapid falling-in and rapid wash-out. The PI value of 8 patients with pancreatic cyst was 0. 181 ±0. 036, with no enhanced blood flow in the cyst. The TIC was a straight line. Conclusion CE-EUS with quantitative analysis is a promising method that can be a more objective basis in the differential diagnosis of pancreatic diseases.

Objective To investigate the lesionsin theleft thalamus and to explore their correlation with executive functionin patients with subcortical ischemic vascular disease(SIVD)by using 1H magnetic resonance spectroscopy (1H-MRS) and diffusion tensor imaging (DTI). Methods Fourteen patients with vascular dementia (VaD), 14 patients with vas-cular cognitive impairment without dementia(VCIND)and 14 normal controls (NC) were recruited. The quantitative analy-sis of N-acetylaspartate (NAA), Choline (Cho), myoinositol (MI) and Creatine (Cr) resonance signals in region of interests (ROI) in the left thalamus were measured. Fractional anisotropy (FA) and mean diffusivity (MD) values in the same region were examined by using DTI. The relationship were analyzedamong the MRS, DTI ratios and cognitive impairment reflect-ed in MMSE and trail making test(TMT). Results The NAA/Cr ratio showed a gradual decrease in the left thalamus of VaD andVCIND patients compared with controls(F=3.656, P=0.038). There was no significant difference in NAA/Cr ra- tiobetween VaD and VCIND patients.Compared with controls, patients with VaD and VCIND demonstrated increased MD value in the left thalamus(F=3.882, P=0.030).There was no significant difference in MD value between VaD and VCIND patients (P>0.05). There were no correlations between NAA/Cr, MI/Cr, Cho/Cr value and DTI parameters in the ROI (P>0.05). TMTb time was positively associated with Cho/Cr(r=0.520,P=0.001 ) and with MD value in the left thalamus (r=0.305, P=0.044). Conclusions Combination of 1H-MRS with DTI can provide the valuable informationon the potential lesions in the thalamus in patients with SIVD. The Cho/Cr and MD values in the leftthalamus may be correlated with ex-ecutive function.

Objective:To explore the clinical features, auxiliary examinations, therapies and prognoses of patients with antibodies targeting glutamic acid decarboxylase 65 (GAD65).Methods:The nine patients with anti-GAD65 neuroimmune disease, admitted to Xuanwu Hospital, Capital Medical University from October 2018 to October 2020, were analyzed, retrospectively.Results:The onset age of nine cases was 17-68 (43.6±20.5) years old, and six cases were female. Two cases had preceding infection. The data of initial symptoms were collected and analyzed, including epileptic onset in four cases, memory impairment in two cases, dizziness in two cases and limb stiffness in one case. As the disease continued to advance, one case developed cerebellar ataxia, one case presented with isolated epilepsy, five cases suffered from limbic encephalitis, one case had stiffman syndrome, one case had brainstem encephalitis. Five cases had antibodies against thyroid peroxidase. Brain magnetic resonance imaging scan showed abnormal signals of T 2/fluid attenuated inversion recovery sequences in four cases, mainly involved bilateral temporal lobes or hippocampus. Epileptiform discharges of frontal or temporal regions of electroencephalography were observed in six cases. All cases received immunotherapy and long-term follow-up was performed in seven cases. Four cases benefited from the immunotherapy. Among the four patients, one fully recovered and returned to work, the other three cases developed neurologic sequelae, including seizures (two cases), and short-term memory loss (one case). The remaining three patients were unresponsive to treatment. Conclusions:GAD65 antibody-mediated neuroimmune disease is a rare neurological disorder, presenting with various syndromes including limbic encephalitis or stiffman syndrome, which is more susceptible to young female. The clinical manifestations included epileptic onset, limb stiffness, cognitive impairment and cerebellar ataxia, etc. Detection of GAD65 antibody in serum or cerebrospinal fluid was gold standard. Early immunotherapy contributed to improving the prognosis of patients, especially for those patients with epileptic onset as the main symptom.