Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.

Trypanosoma is a human parasite severely affecting poor tropical areas.However,current frontline drugs for Trypanosoma treatment have severe side-effects with decreased effectiveness.Based on the fact that aminoacyl-tRNA synthetase is a bonafide drug target for several microorganisms,including bacteria and fungi,it is plausible that it may also be effective target of Trypanosoma.The Trypanosoma brucei leucyl-tRNA synthetase(tbLeuRS)was cloned,expressed and purified to develop an in vitro enzymatic assay system.The assay conditions were further optimized for the effective screening of tbLeuRS inhibitors thus establishing an anti-Trypanosoma drug screening system targeting tbLeuRS.The results indicated that this system can be employed for the effective screening of anti-Trypanosoma drugs with satisfactory specificity.In addition,this system can also be used for compound optimization,as well as IC50 testing.Using this system a series of compounds are identified that are effective Trypanosoma inhibitors without toxicity to human cells.Therefore,targeting tbLeuRS may represent a new venue for the development of anti-Trypanosoma drugs.

Objective To analyze the safety and effectiveness of photoselective green-light laser vaporization of the prostate(PVP)in treatment of large prostate with volume more than 80 ml. Methods Retrospective analysis was performed in 25 benign prostatic hypertrophy patients undergoing the PVP treatment,whose prostate volume was more than 80 ml.Results Twenty-five patients with prostate mean volume of 151.8(84.0-270.0)ml were treated by PVP and 23 cases were finished successfully.One operation was suspended because of bleeding and the other because of damaging orifice.No blood transfusion was needed and no“transurethral resection(TUR)syndrome”happened.Five cases had acute retention and 2 of them needed second PVP.In a mean of 7.9(3.0- 12.0)months follow-up,maximal urinary flow rate(Qmax),international prostate symptom score (IPSS),quality of life score(QOL),and prostate-specific antigen(PSA)level were improved significantly.The volume of prostate reduced after PVP than before PVP but no significant difference was found.Conclusions PVP is feasible in treating large prostate even the volume is more than 80 ml,although it needs better skill.Catheterization for some time is helpful in reducing the incidence of early postoperative acute urinary retention.

OBJECTIVETo study the effects of ultrasound guided inter-scalene brachial plexus block and patient-controlled infraclavicular brachial plexus block for postoperative pain and surgical efficacy in patients with terrible tyriad of the elbow.

METHODSFrom March 2015 to August 2016, 60 patients with terrible tyriad of the elbows were treated in Ningbo No.6 Hospital with ASA I to II internal fixation. There were 32 males and 28 females, ranging in age from 16 to 70 years old, with a mean age of (55.6±18.2) years old. All the patients were divided into two groups(30 cases in each group): controlled intermuscular groove brachial plexus block (group C), infraclavicular brachial plexus block(group I). All catheters were placed using ultra-sound visualization and injected 0.33% ropivacaine 30 ml preoperatively. After regaining consciousness, all patients connected the electronic pump. The solution contained 0.2% ropiva-caine and the pump was setup to deliver a 5 ml bolus dose, with a 15 min lock out interval and background infusion at 5 ml/h. Both analgesia lasted until 5 d after operation. The patients underwent rehabilitation exercise everyday for 5 consecutive days starting from 24 h after operation.VAS score was recorded at 24 h, 48 h, 72 h and 4 d, 5 d after operation during rest and rehabilitation exercise time. The elbow articular range of motion and Mayo elbow performance score (MEPS) were recorded at 6 d after operation. Catheter-related adversereactions (such as oozing from the insertion site, obstruction, prolapse) were recorded.

RESULTSThe success rate of blockade was 100% during insertion in both groups. Compared with group C, the VAS score at 3 d during rest time and 3, 4, 5 d after operation during rehabili-tation exercise were decreased(2.5±0.5 vs. 3.8±1.1, 3.0±0.4 vs. 5.0±0.9, 2.5±0.4 vs. 4.5±1.2, 2.1±0.3 vs. 4.1±1.0,<0.05). The elbow articular range of motion and MEPS were increased(-2.19±18.01)° vs.(-8.19±12.16)°, (45.15±11.20)° vs. (22.15±7.02)°, (19.06±6.75)° vs. (9.10±2.48)°, (17.08±5.18)° vs. (10.12±3.15)°, (80.80±9.50) points vs. (64.90±11.21) points. The incidence of insertion site, obstruction, prolapse was 15, 5 and 10 cases respectively in group C, but without any catheter-related adverse reactions happened in group I (<0.05).

CONCLUSIONSPatient-controlled infraclavieular brachial plexus block can be effectively used for postoperative pain after fixation for terrible tyriad of the elbows, and it can increase surgical outcome.

OBJECTIVETo culture and amplify the young rabbit's bone marrow stromal cells (BMSCs) in vitro, and to observe the effect of hypothermia on the cells' growing behavior and biological function.

METHODSBMSCs were acquired from the rabbit' tibia bone marrow and induced to mature osteoblasts in vitro. The cultured cells growing well in vitro were preserved in liquid nitrogen. The anabiotic cells having cryopreserved for 1 week were chosen as the experimental group, and the routine 7th generation as the control group. Their biological function in comparion by the examination of morphological changes, cells' proliferation ability, colone forming ratio, synthesis ability of ALP and protein, mineralized nodes forming ability were observed.

RESULTSAs contrast to the control groups, the anabiotic cells also grew and proliferated well in vitro except a little more slowly than before. They had the similar general shape in all the time segments, but a little differences in cells' ultrastructure. The experimental groups also had the typical characters of mature osteoblasts, and high abilities of the synthesis of ALP and proteins. The statistic data showed that these two groups had no significant difference (P > 0.05).

CONCLUSIONThe cryopreserved osteoblasts had the same biological functions and the similar growing behaviors as before. These results suggest that it is practical to use the cryopreserved osteoblasts for further study on bone tissue engineering.

Animals ; Bone Marrow Cells ; Bone and Bones ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; In Vitro Techniques ; Osteoblasts ; Rabbits ; Tissue Engineering

AIMTo investigate human epidermal growth factor receptor type 2 (HER2) protein expression and gene amplification in Chinese metastatic prostate cancer patients and their potential value as prognostic factors.

METHODSImmunohistochemistry (IHC) was performed to investigate HER2 protein expression in prostate biopsy specimens from 104 Chinese metastatic prostate cancer patients. After 3-11 months of hormonal therapy, 12 patients underwent transurethral resection of the prostate (TURP). HER2 protein expression of TURP specimens was compared with that of the original biopsy specimens. Of these, 10 biopsy and 4 TURP specimens with HER2 IHC staining scores >or=2+ were investigated for HER2 gene amplification status by fluorescent in situ hybridization (FISH).

RESULTSOf the 104 prostate biopsy specimens, HER2 protein expression was 0, 1+, 2+ and 3+ in 49 (47.1%), 45 (43.3%), 8 (7.7%) and 2 (1.9%) cases, respectively. There was a significant association between HER2 expression and Gleason score (P = 0.026). HER2 protein expression of prostate cancer tissues increased in 33.3% of patients after hormonal therapy. None of the 14 specimens with HER2 IHC scores >or= 2+ showed HER2 gene amplification. Patients with HER2 scores >or= 2+ had a significantly higher chance of dying from prostate cancer than those with HER2 scores of 0 (P = 0.004) and 1+ (P = 0.034). Multivariate Cox regression analysis showed that HER2 protein expression intensity was an independent predictor of cancer-related death (P = 0.039).

CONCLUSIONAn HER2 IHC score >or= 2+ should be defined as HER2 protein overexpression in prostate cancer. Overexpression of HER2 protein in cancer tissue might suggest an increased risk of dying from prostate cancer. HER2 protein expression increases in some individual patients after hormonal therapy.

Aged ; Antineoplastic Agents, Hormonal ; therapeutic use ; Asian Continental Ancestry Group ; genetics ; statistics & numerical data ; Biopsy ; China ; epidemiology ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Prostatic Neoplasms ; drug therapy ; genetics ; mortality ; secondary ; Receptor, ErbB-2 ; genetics ; metabolism ; Risk Factors

OBJECTIVETo investigate the efficiency and safety of transperitoneal laparoscopic adrenalectomy for the treatment of adrenal tumors and to describe surgical technique and management of intraoperative complications.

METHODSFrom February 2002 to April 2008, 24 male and 44 female consecutive patients with average age (53.4 +/- 12.1) years old were treated with transperitoneal laparoscopic adrenalectomy because of adrenal tumors. Of the patients, 27 cases had right adrenal tumors, 39 cases had left adrenal tumors and 2 cases had bilateral adrenal tumors. The average tumor size was (2.9 +/- 2.0) cm with the maximal diameter of 10 cm. We evaluated this technique in respect of operating time, estimated blood loss, complications during surgery, postoperative complications, duration of hospital stay and pathological results.

RESULTSAll the surgeries had been completed successfully without conversion to open surgery. The average operating time was (157.7 +/- 51.5) min, the average estimated blood loss was (68.1 +/- 54.2) ml. No major complication happened during operation. The average drainage time was (2.6 +/- 1.5) days. The average post-operative hospital stay was (8.7 +/- 4.3) days. Four cases (6.0%) developed surgical field liquefaction and 2 cases (2.9%) experienced delayed closure of the drainage wound.

CONCLUSIONSTransperitoneal laparoscopic adrenalectomy is feasible and safe in the treatment of adrenal tumor with low risk of intra-operative and post-operative complication.

Adrenal Gland Neoplasms ; surgery ; Adrenalectomy ; methods ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVESTo analyze the epidemiology information of prostate cancer from three centers of Beijing, Shanghai, Guangzhou, and to reflect the current situation of prostate cancer in China, and to analyze the information of 272 patients with advanced prostate cancer who received hormonal therapy to find the prognostic factors of hormone therapy.

METHODSCollect the information of 525 patients with prostate cancer from three centers. Two hundred and seventy-two cases of advanced prostate cancer with full information were selected from the 525 cases to analyze the prognostic factors of hormone therapy.

RESULTSThree hundred and fifty-seven cases (68.0%) had advanced disease at diagnosis and 80.2% patients received hormone therapy as the main therapy. Prognostic analysis indicated that Gleason score, bone metastasis and prostate specific antigen nadir were independent prognostic factors of progression-free survival time.

CONCLUSIONSIn this report, most patients are advanced prostate cancer at diagnose, and hormonal therapy is the main therapy. Gleason score, bone metastasis, prostate specific antigen nadir are independent prognostic factors of advanced prostate cancer after hormone therapy.

Adult ; Aged ; Aged, 80 and over ; Androgen Antagonists ; therapeutic use ; Antineoplastic Agents, Hormonal ; therapeutic use ; China ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Prostatectomy ; Prostatic Neoplasms ; epidemiology ; pathology ; therapy ; Survival Analysis

OBJECTIVETo investigate the efficacy of repeat transurethral resection of tumor in patients with non-muscle invasive bladder cancer.

METHODSFrom March 2004 to August 2008, 462 patients (350 males, 112 females, aged from 35 to 83 years old) with non-muscle invasive bladder cancer, were evaluated according to tumor stage, grade and muscle or no muscle tissue in initial transurethral resected sample. One hundred and twenty-five patients underwent repeat transurethral resection of bladder tumor within 4 to 6 weeks after initial resection. Of these 125 patients 49 were Ta, 76 were T1, 58 were low grade carcinoma, 67 were high grade carcinoma and 30 were not found presence of muscle tissue in initial resected sample in patients with T1 stage.

RESULTSOf the 125 cases, 34.4% (43/125) had residual tumor and 65.6% (82/125) had no tumor on repeat transurethral resection. Of 43 cases with residual tumor 35 had non-muscle invasive tumor including 15 in Ta and 20 in T1. The patients with high grade carcinoma had more residual tumor than those with low grade carcinoma (P < 0.05). The patients with muscle tissue in initial transurethral resected sample had fewer residual tumor than those without (P < 0.05). Twelve cases (9.6%) were understated at initial resection. Six cases (4.8%) had bladder perforation and 7 (5.6%) had bleeding during repeat transurethral resection. All cases were followed up for 3 to 56 months (median 26 months), 37.2% (16/43) patients with residual tumor in repeat transurethral resection had recurrence while only 12.2% (10/82) without residual tumor in repeat transurethral resection did (P < 0.05).

CONCLUSIONSRoutine repeat transurethral resection is advised to non-muscle invasive bladder cancer patients with T1 tumor or high grade carcinoma or no muscle tissue in initial transurethral resected sample within 4 to 6 weeks after initial resection. Repeat transurethral resection could increases the stage accuracy.

Adult ; Aged ; Aged, 80 and over ; Electrosurgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Reoperation ; Retrospective Studies ; Treatment Outcome ; Urinary Bladder Neoplasms ; surgery ; Urologic Surgical Procedures ; statistics & numerical data

Objective To analyze the changes of the main clinic parameters in patients with benign prostate hyperplasia (BPH) treated by watchful waiting and to find out the risk factors contributing the progress of BPH. Methods According to the inclusion and exclusion criteria, 61 patients diagnosed as BPH were recruited in the group of watchful waiting. Data on IPSS, prostate volume, prostate specific antigen(PSA), maxium flow rate, average flow rate and residual urine volume during follow-up period of 24 months, were recorded. Results At 0, 12, 24 months, the IPSS, prostate volume (ml), PSA(ng/ml),maxium flow rate (ml/s) were 7±4, 4±3, 4±3 ; 33.0±9.0, 33.8±7.6, 30.9±6.8 ; 1.53±1.35,1.43±0.95, 1.22±0.99; 17.1±5.0, 17.2±6.1, 19.2±8.0, respectively. At the end of the 24-months follow-up, all observed parameters had a little improvement except the average prostate volume in this group. Of the 61 patients, 42(62%) progressed slowly or became better when comparing with baseline data of the study. Moreover, the difference between at 24-month and at baseline period, IPSS showed statistical significance (P＜0.0001) in t test. In the study of BPH progression risk factors by logistic regression analysis, prostate volume( P = 0. 0910) and residual urine volume( P = 0. 0780) showed a trend of becoming the risk factors. Conclusion Our study showed that patients treated with watchful waiting had slow progression and majority of these patients did not need to alter their treatment options.Through data analysis, we noticed that the changes of data watchful waiting patients could help us to choose more precise and reasonable treatment option in clinical pratice.[ Key words ] Benign prostate hyperplasia; Prostate volume; Residual urine; Watchful waiting;Follow-up