ObjectiveTo explore the clinical effects of laparoscopic common bile duct exploration and primary suture combined with choledochoscopy (LBEPS) and laparoscopic common bile and duct exploration T-tube drainage(LCHTD) on cholelithiasis.Methods101 cholelithiasis patients were selected and,grouped by the operation methods.56 cases were treated with LCHTD,while 45 cases were treated with LBEPS.The operation time,blood lose,and postoperative digest function recovery time were ampared betreen two groups.ResultsThe operation time betreen the two groups has no significant difference(P ＞ 0.05 ),while the blood lose,postoperative digest function recovery time in LBEPS group were better than the LCHTD group( all P ＜ 0.05 ),the differences were statistically significant.Meanwhile the LBEPS group had a less complication and re-treatment rate than those of the LCHTD group ( P＜0.05).ConclusionCompared with LCHTD,LBEPS had smaller trauma,less operation complications and faster postoperative digest recovery time,worthy of clinical promotion.

Objective To analyze the factors of Protein A/G affinity column,influencing purification effect,and get the best condition to improve application effect of Protein A/G affinity column. Methods We Purified anti-HCV-IgG serum with different disposal modality,different sample input modality and different application number of affinity column before detecting and analyzing the purified samples. Results The Protein A/G affinity column had the best purified effect after using saturated ammonium sulfate to first purification,which increased the affinity column adsorption effect within 30 minutes adsorption. Conclusion Using antibody with first purification and adding the adsorption time could improve utilization rate of affinity column and prolong the service life.

Objective To optimize the PCR primer sets for Simian virus 40 (SV40) detection and establish an assay method for SV40 which is of high sensitivity, strong specificity, broad applicability. Methods Two pairs of PCR primers were designed of based on 21 different SV40 strains genome by Primer Premier 5.00 software, and the features of two pairs of PCR primers were analyzed by Oligo software (version 6.71), conservative nucleotide of two pairs of PCR primers and the PCR amplification product were analyzed by DNAMAN software (version 6.0.40). Two pairs of new-built PCR primers were compared with those derived from China pharmacopoeia (Clip) in these aspects. The detection sensitivity of four pairs of PCR primers were analyzed using different SV40 DNA diluent as PCR template. The detection specificity of four pairs of PCR primers were analyzed using sterile water, Vero cell DNA, SV40 DNA as PCR template, respectively. Results The sequences of the new PCR primer sets VP1 and T are conservative for 21 Strains. The sequences of PCR primer sets GCVP1 and GCT are conservative for SV40 strains whose accession No. is J02400, NC_001669, AF316139 and AF316141. As far as the same diluent SV40 DNA template is concerned, the PCR amplification efficiency of PCR primer set VP1 and T is higher than that of PCR primer set GCVP1 and GCT. There are non-specific band in nucleic acid electrophoresis for amplification products of PCR primer sets GCVP1 and GCT, whereas there are no non-specific band in nucleic acid electrophoresis for amplification products of PCR primer sets VP1 and T. Conclusion The new assay method for SV40 nucleic acid sequence has many better qualities than those in Chp such as high sensitivity, strong specificity, broad applicability, conservation of primers and their amplification products and so on.

Objective To investigate the efficacy of CT guided radioactive 125I seed implantation on elderly patients with advanced malignancies.Methods 78 cases of elderly patients with malignant were collected and divided into two groups.41 cases of the treatment group were treated with CT guided radioactive 125I seed implantation.Particle activity was 29.6 MBq.The prescription dose was 90-110 Gy.37 cases of the control group were treated with optimized supportive care.Data from all of patients were to review and followup observation in short term efficacy,quality of life and side efforts.Results The total effective rate was 92.7 ％(38/41)and the disease control rate was 97.6 ％(40/41)in the treatment group.The control group was in the effective rate and 16.2 ％(16/37)in the disease control rate.The quality of life of the treatment group was higher than those in the control group(P＜0.05).And there is no obviously side efforts.Conclusion The treatment of elderly patients with advanced malignant tumors by 125I seed implantation was a safe and effective method.It can improve the quality of patients' life.

Objective:To evaluate the efficacy of computed tomography (CT)-guided implantation of radioactive ~(125)I seeds in the treatment of residual foci of nasopharyngeal carcinoma after radiotherapy.Methods:Twenty-five nasopharyngeal carcinoma patients who had residual foci after radiotherapy were implanted radioactive ~(125)I seeds from January 2007 to January 2008 in our hospital. Three-dimensional treatment planning system (TPS) was used to calculate the quantity and total dosage of ~(125)I seeds. The radioactive ~(125)I seeds were implanted into residual foci under CT guidance. The dosage distribution of radioactive ~(125)I seeds were verified after surgery. The nasopharyngeal endoscopy CT scans were performed at 3, 6, and 12 months after surgery and the short-term efficacy and adverse reaction were evaluated. Results:Out of the 25 patients, 20 patients had complete response (CR), 2 patients had partial response (PR), 2 patients had no change (NC), and 1 patient had progressive diseases (PD). The overall effective rate (CR+PR) was 88.0%. All patients had no serious postoperative complications. Conclusion:CT-guided implantation of radioactive ~(125)I seed was an effective and safe method in the treatment of the residual foci of nasopharyngeal carcinoma after radiotherapy.

ObjectiveTo evaluate the primary effect of granulocyte-monocyte colony stimulating factor (GM-CSF) as an immunotherapy option for treatment of residual disease after alloHSCT.Methods Immunotherapy was performed on two patients with blood malignancy to treat residual disease after allo-HSCT. The patient one,who was diagnosed as having MDS-RAEB Ⅱ,showed bone marrow displasis and incomplete chimerism 6 months after unrelated donor HSCT.Immunosuppressive drug was withdrawn without induction of graft-versus-host disease (GVHD).The patient two B-ALL demonstrated a residual disease at molecular level 30 days post-transplantation.Both of them were given GMCSF (300 μg) subcutaneously once every two days for totally three weeks.During the whole period,skin itch and rash,liver function,subgroups of lymphocytes,and MDSCs and DCs in peripheral blood were investigated.Results In case one,grade Ⅰskin acute GVHD (aGVHD) appeared as early as one week after GM-CSF administration,as well as grade Ⅱ (skin and liver) by the end of the third weeks,and GM-CSF injection was withdrawn.One month later since the start of GM-CSF,the patient showed normal bone marrow morphology and full donor type chimerism. Cyclosporine A (CsA), mycophenolate mofetil and methylprednisolone were administered for two weeks to control GVHD.In the other case,grade Ⅰ aGVHD occurred 9 days after GMCSF administration,and whole blood CsA maintained at 0.134-0.472 μmol/L.Prednisone (30mg per day for 5 days) was used to control grade Ⅱ GVHD from the 11th day after GM-CSF,and grade Ⅰ GVHD continued without any intervention.On the 30th day after GM-CSF treatment,bone marrow aspiration showed complete molecular remission.In both of the two cases,no differences in lymphocytic subtypes were revealed before and after GM-CSF administration,while there were trends of increased DC number and decreased MDSCs in peripheral blood.ConclusionThe administration of GM-CSF as an immunotherapy option for blood malignancy may contribute to the clearance of residual disease after Allo-HSCT.

Objective:To observe effect of Xiaoyu Zhixue Tablets on expression of platelet membrane glucoprotein in the patient of hemorrhagic thrombopathy and probe into the mechanism of the therapy.Methods:148 cases of hemorrhagic thrombopathy were randomly divided into a Chinese drug group(n=98)treated by Xiaoyu Zhixue Tablets,and a Western medicine group(n=50) treated by adrenosem,vitamine C,K,P.They were treated for 6 months.After treatment the therapeutic effect of hemostasis and the recovery rate of platelet aggregation in the two group were observed and analyzed.Before and after treatment expressions of platelet membrane glucoproteins GP Ⅰ b/Ⅸ,GP Ⅱ b/Ⅲa,GP Ⅰ b,GP Ⅱ b,GP Ⅲ a and p-selectin expression were detected with flow cytometry in the two groups and in 34 healthy persons(normal group).Results:The total effecive rate of hemostasis was 89. 8% in the Chinese drug group and 54.0% in the Western medicine group,and the recovery rate of platelet aggegation was 72.4% in the Chinese drug group and 4.0% in the Western medicine group,with significant differences(both P0.05).Conclusion:Xiaoyu Zhixue Tablets can up-regulate platelet membrane glucoproteins GP Ⅰ b/Ⅸ,GP Ⅱ b/Ⅲ a,GP Ⅰ b,GP Ⅲ a and p-selectin expression,which is possibly one of the mechanisms for treatment of hemorrhagic thrombopathy.

To explore the possibility and condition of differentiation of bone marrow mesenchymal cells (BMSCs) to neural cells in vitro, BMSCs from whole bone marrow of rats were cultured. The BMSCs of passage 3 were identified with immunocytochemical staining of CD44 ( + ), CD71 ( + )and CD45(-). There were type Ⅰ and type Ⅱ cells in BMSCs. Type Ⅰ BMSCs were spindleshaped and strong positive in immunocytochemical staining of CD44 and CD71, whereas flat and big type Ⅱ BMSCs were lightly stained. The BMSCs of same passage were induced to differentiate into neural cells by β-mercaptoethanol (BME). After induction by BME, the type Ⅰ BMSCs withdrew to form neuron-like round soma and axon-like and dendrite-like processes, and were stained positively for neurofilament (NF). The type Ⅱ BMSCs did not change in the BME medium and were negatively or slightly stained of NF.

Objective To analyze the biological characteristics of clinical isolates of coxsackievir-us A6 (CVA6), a pathogen of hand,foot and mouth disease (HFMD), and to provide reference for vaccine development. Methods CVA6 strains were isolated from 21 stool and throat swab specimens of patients with HFMD in Yunnan Province and then identified. Their growth characteristics, plaque morphology and virulence to suckling mice were analyzed. Results Five CVA6 strains, named CVA6-129, CVA6-113, CVA6-57, CVA6-94 and CVA6-162, were isolated and all belonged to D3 subtype. Only the CVA6-129 strain could proliferate rapidly in Vero and KMB17 cells and the proliferation peaked 30 h after inoculation. The infectious titer of the CVA6-129 strain was 7. 54 lgCCID50 (50％ cell culture infective dose) / ml in KMB17 cells. Different morphologies of plaques were formed by the CVA6-129 strain in Vero and KMB17 cells at the same time points, which were small and round with clear edges in Vero cells, and large and irregular with blurry edges in KMB17 cells. Suckling mice were susceptible to CVA6 via intramuscular and intraperito-neal injection. The most common symptoms in infected suckling mice were reduced mobility, hind limb pa-ralysis and quadriplegia. CVA6 infection could result in death in severe cases. Conclusions This study isolated five CVA6 strains from a number of clinical samples of suspected HFMD cases, of which the CVA6-129 strain showed potential as a vaccine candidate.

【Objective】 To analyze the reasons for the invalidity of blood nucleic acid test results, and to explore the countermeasures to reduce the invalidity of the test. 【Methods】 From 2019 to 2021, the number of tests performed in our laboratory for Cobas s201 blood nucleic acid screening system and the number of batches and tests with invalid results were counted, and the types and reasons of invalid results were analyzed. 【Results】 From 2019 to 2021, the Cobas s201 nucleic acid detection system detected a total of 5, 420 batches and 127, 950 pools, and the invalid rate of batches and pools were 1.83% and 1.97%, respectively. The types of invalid results can be summarized as improper operation, sample quality problems, invalid quality control (IQC), equipment failure and others. Among them, IQC and equipment failure were the main reasons for invalid results, accounting for 44.51% and 39.96%, respectively. IQC was mainly related to cross-contamination of samples and insufficient mixing of quality control products. Equipment failures mostly occurred in the robotic arm gripper of the nucleic acid extraction instrument and the TC module of the amplification instrument. 【Conclusion】 The laboratory should conduct quality monitoring for invalid results, and take targeted improvement measures, especially to reduce invalid results caused by invalid quality control and instrument failure.