Objective To investigate the clinical efficacy and safety of the treatment of type 2 diabetes mellitus with Humalog 25R and Humulin 70/30.Methods120 patients with type 2 diabetes were collected in The first people's Hospital of Jiande in the 3 years, the cases of patients were randomly divided into control group (60 cases) and observation group (60 cases);for the patients in the control group were treated with Humulin 70/30, the patients in the observation group were treated with Humalog 25, compared the experimental data between the two groups.ResultsThe clinical symptoms of the observation group were better than those of the control group after treatment, the clinical effect of the observation group was better than that of the control group, the difference was statistically significant (P<0.05).ConclusionThe treatment of type 2 diabetes patients with Humalog 25 can effectively improve the clinical symptoms of patients, the treatment effect is good, worthy of clinical application.

Objective To introduce an optimal incision technique in the endourethral surgery for urethratresia. Methods Sixteen patients suffering from urethratresia were managed by endourethral surgery,2 of them treated with rotator incision and the other 14 with rotator incision plus transurethral scar electrosectomy.The efficacy of the treatment was evaluated by Qmax. Results Successful incisions were achieved in all patients,the duration of procedure being from 18 to 33 min,with a mean of 25 min.Two patients with rotator incision had dysuria (Qmax 9 and 7 ml/s) after the surgery and accepted transurethral resection of scar again.The other 14 patients with rotator incision plus transurethral scar electrosectomy had good voiding.Qmax was more than 16 ml/s in all patients after operation. Conclusions Rotator incision is a safe,simple,efficacious incision technique in the endourethral surgery for urethratresia,and scar electrosectomy is essential for successsful outcome.

Objective:To study the relationship betwen the mechanism of autoimmune disease and CD4+CD25+ T cell population in NIK mutated mice-aly mice.Methods:NIK mutated mice-aly/aly mice were used as model,aly/+mice as NIK normal control;cell populations were determined by FACS and the thymus structure were analyzed by immunohistochemistry.Results:The CD4+CD25+CD8- population were remarkably decreased in aly mice;and the UEA-1 positive cells were absent in aly mice.Conclusion:The autoimmune disease in aly mice might be the result of deceased the CD4+CD25+ population;the UEA-1 positive cells might play an important role in the development of CD4+CD25+ population. [

Tethered cord syndrome is a severe and congenital disease. Resecting the diseased ilium terminal(FT), eystis, lipoma is the main treatment for the disease, relieving the injury to the eonus medullaris. Why the FT is diseased and thickened? Is it necessary to eut internal FT or external FT? What is the injury mechanism of the low-positional or normal positional oonus medullaris? To study the component and structure of the conus medullaris and FT may give definite answers.

Objective To analyze the efficacy, safety, and learning curve of holmium laser enucleation of prostate (HoLEP). Methods From June 2014 to June 2015, there were 45 consecutive patients with benign prostatic hyperplasia who underwent HoLEP performed by one experienced urologic doctor ( having surgical experience of transurethral resection of prostate in more than 500 cases) in our hospital. The patients were divided into three groups:group A (patient 1-15), group B (patient 16-30), and group C (patient 31-45). The patient age, prostate specific antigen (PSA) and prostatic volume evaluated by ultrasound were similar among the 3 groups (P>0. 05). Clinical data including operative time, hemoglobin decrease, and catheterization duration were compared among the 3 groups. The efficiency for enucleation was calculated as prostatic volume divided by operation time. Results The average enucleation efficiency of 3 groups was 0. 34 ± 0. 14, 0. 36 ± 0. 16, and 0. 49 ± 0. 18 ml/min, respectively, with a significant difference among the 3 groups (F =4. 025, P =0. 025). The group C obtained significantly improved efficiency as comparing with the group A and group B (P=0. 013 and 0. 028, respectively). No significant difference was found in operative time, hemoglobin decrease, catheterization duration, and rate of blood transfusion and complication among the 3 groups (P>0. 05). There were several surgery-related complications, including 2 cases of conversion to TURP, 5 cases of postoperative temperature more than 38℃, 4 cases of severe stress incontinence, 1 case of bladder mucosal injury by morcellator, 1 case of urethral stricture, and 1 case of postoperative hemorrhage. It took 30 cases to overcome learning curve of HoLEP by self-taught modality. Conclusions For experienced endourologist, HoLEP can be learn by self-taught modality after 30 cases training. Cases with small prostatic volume less than 60 ml should be selected in the early stage of the learning curve.

Objective To evaluate the effect of duration of subcutaneous injection on the extent of bruising at injection sites among patients receiving low- molecular weight heparin. Methods Randomized clinical trials and quasi-experimental within-subject design that assessed the effect of subcutaneous duration on the bruising of injection sites were selected in Cochrane library, JBI evidence-based practice medical and nursing database, PubMed biomedical information retrieval platform, Medline general medical literature retrieval database. RevMan5.3 software was used to analyze the data after quality assessment, and data extraction were made for eligible trials. Results A total of 9 trials were included. The results of Meta-analysis supported the 30 s duration injection technique on reducing the rate of bruising of injection sites than 10 s duration injection technique (odds ratio was 0.34,95%confidence interval 0.25-0.47, P<0.01).While it was failed to support the effect on reducing the bruising area 48 and 72 hours after injection (P > 0.05). Conclusions Increasing the length of low-molecular weight heparin subcutaneous injection can reduce the rate of bruising. While it could not effectively reduce the area.

Objective To study the risk factors of central lymph node (CLN) metastasis in papillary thyroid carcinoma (PTC),to provide the basis for clinical treatment.Methods A total of 407 patients with PTC in thyroid surgery,the Third Affiliated Hospital of Harbin Medical University from January 2010 to June 2012 who had undergone at least ipsilateral CLN were studied.These factors included normal situation,thyroidglobulin antibody or thyroid microsomal antibody,family history,pathological features (the size of primary,tumor multifocal,tumor location and capsular invasion),TNM staging and CLN metastasis.Univariate analysis and multivariate analysis were performed using x2 test and binary Logistic regression test,respectively.Results The CLN metastasis rate was 51.351％ (209/407) of the 407 patients with PTC.The CLN metastasis was significantly associated with age,tumor multifocal,the size of primary tumor,capsular invasion and TNM (x2 =21.080,14.974,47.671,12.858,8.765,all P ＜ 0.01).Sex,thyroid globulin antibody or thyroid microsomal antibody,family history and tumor location were not associated with CLN metastasis (x2 =1.457,1.106,0.000,0.260,all P ＞ 0.05).Multivariate analysis indicated that age (＜ 45 year),the size of primary tumor (≥ 1.0 cm),tumor-muhifocal,and TNM (T3 + T4) were the independent risk factors for CLN metastasis [odds ratio (OR) =0.937,2.347,0.380,0.389,all P ＜ 0.01].Conclusions Age (＜ 45 year),the size of primary tumor (≥ 1.0 cm),tumor-multifocal and TNM (T3 + T4) are risk factors for CLN metastasis in PTC.Dissection of CLN should be considered for PTC patients with these factors.

Objective To culture the acute leukemia cells in vitro,and to prepare cancer vaccine expressing heat shock protein 70 (HSP70) of Bacille Calemette-Geérin(BCG) onto the cell surface,so as to study its anti-tumor effect and mechanism.Methods Acute myeloid leukemia (AML) cells were cultured in a serum-free Stemspan(H) culture supplemented with cytokines [stem cell factor(SCF),flt-3 ligand (FL),interleukin (IL)-3 and IL-6] in vitro.And B-lineage acute lymphoblastic leukemia (B-ALL) cells were cultured in a Iscove modified medium(IMDM) culture supplemented with cytokines (SCF,FL,IL-3 and IL-7) in vitro.Cellular morphology was observed by the microscopy and immunophenotype determination was used to verify the biological characteristics of acute leukemia cells after culture.Lipofectamine 2000 was used to transfect the pDisplay-HSP70 plasmid into acute leukemia cells.The expression of HSP70 on the cell surface was detected by fluorescene microscope.Then the immunogenicity of the leukemia cells expressing HSP70 were detected.The experimental groups were divided into 3 subgroups:the wide-type acute leukemia cells (wt-LC group),the pDisplay-leukemia cells (pDisplay-LC group),and the pDisplay-HSP70-leukemia cells (HSP70-LC group),respectively.The leukemia cells in different groups were cultured with autologous peripheral blood T cells for 72 hours.The proliferation indices of T cells were assayed by carboxyfluorescein diacetate succinimidyl ester (CFSE)-staining method,and the contents of interferon-γ(IFN-γ) were tested by enzyme-linked immunosorbent assay (ELISA).The leukemia cells in different groups were cultured with autologous peripheral blood T cells,and after 6 days,the fresh acute leukemia cells were added [in the different ratios of cytotoxicity T lymphocyte (CTL):leukemia cells were 10 ∶ 1,20 ∶ 1,40 ∶ 1 and 80 ∶ 1] and continued to be cultured for another 12 hours.Cytotoxicity assay was measured by lactate dehydrogenase (LDH) release.Results After short term culture in vitro,the leukemia cells were in colony-like suspension and maintain the proliferation characteristics were maintained.The cell proliferation was rapidly cultured for about 10 days and then was gradually slowed down.But there was no difference between the day 10 and day 0 in the expressions of CD13 and CD33 in fifteen cases of AML cells (P ＞ 0.05).Equally,there was no difference between the day 10 and day 0 in the expressions of CD19,CD10 and CD22 in fifteen cases of B-ALL cells (P ＞ 0.05).After BCG HSP70 gene transfection,the yellow-green fluorescence on the leukemia cells surface was observed under the confocal microscope.Detection of the immunogenicity:(1) Autologous T cell proliferation:the most significant T cell proliferation was observed in the group of HSP70-transfected leukemia cells (t =17.89,19.58,all P ＜0.05).There was no difference between the wt-LC group and pDisplay-LC group (P ＞ 0.05).(2) The contents of cytokines:the IFN-γ level in the group of HSP70-transfected leukemia cells was higher than those of wide-type acute leukemia cells and the pDisplay-transfected ones (t =24.72,24.81,all P ＜ 0.05).(3) Cytotoicity of CTL:the killing rate in HSP70-transfected leukemia cells was significantly higher than those of wide-type acute leukemia cells and pDisplay——transfected ones(F =13.66,P ＜ 0.05).And with the increase of the ratio from 10 ∶ 1 to 80 ∶ 1,the inhibiting activity of CTL in the HSP70-LC group was raising(F =19.69,P ＜ 0.05).Conclusions Fresh acute leukemia cells can be successfully cultured in vitro.Short-term culture can significantly increase the number of leukemia cells,but has little effect on surface antigen expression.So,the biological characteristics of the leukemia cells can be maintained.The leukemia cells vaccine expressing BCG HSP70 onto its surface was successfully prepared,and gene transfection of BCG HSP70 can significantly enhance the immunogenicity of leukemia cells.

Objective To present the primary effectiveness and morbidity of metallic ureteral stent in treating the patients with malignant ureteral obstruction .Methods The retrospective study enrolled 11 patients received metallic ureteral stent placement because of malignant ureteral obstruction at our hospital between Jul.2012 and Jan.2014.Total of 16 stents were inserted including 5 bilateral stent insertion. There were 4 males and 7 females, with a mean age of 56 years (range 28 to 88 years).Mild to moderate hydronephrosis were identified by ultrasound and CTU in all patients , including 2 recurrent fever and 1 renal colic in affected side , however serum creatinine level is normal .There were 10 patients received stents insertion through cystoscopic or nephroscopic retrograde approach and 1 through nephroscopic antegrade approach,under superficial , spinal or intravenous anesthesia .Patients were scheduled for follow-up at 6 months intervals by KUB and ultrasound and would be examined ahead of time if there were any obstructive symptoms including recurrent fever or renal colic of the affected side .Outcomes and complications were observed during follow-up.Results In total,11 patients underwent 13 operations,including 2 by staging operations.Hydronephrosis , recurrent fever and renal colic were relieved in all patients after operation , showing 100%success rate.The mean follow-up was 9 months,in which,9 patients had not reached the scheduled time (1 year) while 2 patients exceed the time limit of stent exchange because of poor physical status.During the follow-up, 1 patient died of the progression of urothelial carcinoma .Nine stents of 5 patients keep alleviating the obstruction without hydronephrosis evaluated by ultrasonography .Stents failure occurred in 6 ureters of 5 patients, including 3 encrustation and 2 migration, presenting of progressive hydronephrosis and recurrent fever . Functional restoration was achieved in one of the encrusted stent undergoing ESWL .One stent with encrustation fail to be removed and a polymer double-J stent replacement in the same side was required for drainage .The other 3 nonfunctional stents were removed and replaced by polymer double-J stents.Conclusions Ureteral obstruction secondary to extrinsic compression contributing to relatively stable malignant tumor could be treated safely and sufficiently with the resonance metallic stent . Surveillance on schedule and regularly change within 1 year is critical to achieve successful outcomes .Tumor progression with ureteral invasion as while as encrustation can cause stent failure ,and in these cases the stent should be changed every 6 months or less.

Objectives To prepare the HL-60 cell vaccine expressing heat shock protein 70 (HSP70) of Bacille calmette-Guérin (BCG), so as to study its anti-tumor effect and mechanism. Methods The whole BCG HSP70 gene was amplified from BCG genome by polymerase chain reaction (PCR) and sub-cloned into the polyclone endonuclease sites in pDisplay. The recom-binant vector of pDisplay-HSP70 was verified by sequencing. Then the HL-60 cell vaccine expressing the protein onto the cell surface was prepared by lipofectamine transfection. To detect the immunogenicity of HL-60 cells expressing HSP70, the test groups were divided into three subgroups, HL60-wt, HL60-pDisplay, and HL60-HSP70 respectively. Each group was cultured with peripheral blood T cells for 72 h, then the proliferation indices of T cells were assayed by CFSE-staining method, and IFN-γwere tested by enzyme-linked immunosorbent assay (ELISA). The HL-60 cells of different groups were cultured with peripher-al blood T cells for 6d. The wild-type HL-60 cells were added and co-cultured for another 12h. Cytotoxicity assay was measured by LDH release. Results (1) The fragment of BCG HSP70 was consistent with the theoretical value. DNA sequencing showed that the recombinant vector of pDisplay-HSP70 was correctly constructed. (2) BCG HSP70 expressed onto the HL-60 cells sur-face. (3) Detection of the immunogenicity: ①The most significant T cell proliferation was observed in the group of HSP70-transfected HL-60 cells (P<0.05). There was no difference between the HL60-wt group and HL60-pDisplay group (P>0.05).②The contents of IFN-γof the HSP70-HL60 group was the highest.③The inhibiting activity of CTLs on HL-60 cells in the group of HSP70-transfected HL-60 cells was more significant than that of wide-type and pDisplay--transfected HL-60 cells. And with the increase of the E:T ratio, the inhibiting activity of CTLs in the HSP70-HL60 group was rising. Conclusions The recombinant eukaryotic expression vector (pDisplay-HSP70) of BCG HSP70 was successfully constructed. And the HL-60 cell vaccine expressing BCG HSP70 onto its surface was successfully prepared. The results showed that gene transfection of BCG HSP70 could significantly enhance the immunogenicity of HL-60 cells.