Objective To search for a good method for generation of hematopoietic stem cells (HSC) by embryonic stem cells (ESC),and to investigate the potential of HSC derived from ESC to reconstruct hematopoiesis in vivo.Methods Using a three-step method to induce a mice ESC line, E14.1-into HSC.And identifying HSC by flow cytometry analysis cell markers with CD34 +/Sca-1+, then HSC (1×109L-1)from ESC differentiation were injected into severe combined immunodefieency (SCID) mice for observing teratoma formation.To validate function of HSC by colonngenic cells assay and to reconstitute the hematopoiesis in lethally irradiated mice.Results Combining to use more hematopoietic stimulating factor to availably promote the El4.1 cell into embryoid body (EBs) with abundant hematopoietic progenitors.EBs were induced after 14 day to fast differentiate for HSC with peak percentages of CD34+/Sca-1+ cells reached to (13.72±2.07)%.To harvested ceils from EBs by day 14 for second -step HSC differentiation, percent of CD34+/Sca-1+ cells rise to(24.62±2.50) % after day 16 induction.Cloning forming units (CFU) analysis showed that more Erythro -myeloid cloning generation was observed at this period.Cells obtained in the second step are subsequently plated on monolayer of mice bone marrow stromal cells, in the presence of TPO, FLt3 ligand and superoatant of mice fetal liver stromal cells, cultured for additional 15 days, followed fast expansion of CD34+/Sca-1+ to maximally (58.64±4.20 ) % with more CFU-E, CFU-GM and CFU-GEMM population.Wright-Giemsa stain showed that its had the character of hematopoietic progenitors.Cells from the third-step were injected into SCID mice, but no teratomas were recovered in 2 mices after 6 weeks.Positive selection of CD34+/Sca-1+ cells by magnetic sorting from the third - step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 days after transplantation, with 71.4% successful engraftment rate.And 3 recipients showed that the cell population of the peripheral blood leukocytes,red cells, hemoglobin approached normal index at 40 days after transplant, hut followed relative'slow renew in platelet count.Survival rate of transplant group is 43.0%, compared to 100% mortality in control mice.Karyotyping assays confirmed female mice with XY.Conclusions The results showed that the three-step differentiation and the culture conditions described here good support differentiation of ESC into HSC.HSC derived from ESC were safe without teratomas formation in body, and its can reconstruct hematopoiesis.

Objective To investigate the clinical significance of contrast-enhanced transrectal ultrasound(CE-TRUS) in the perineal prostate biopsy. Methods A total of 116 patients was undergone prostate biopsy through the perineum under the direction of tansrectal ultrasound. Prostate biopsy standard was based on 2007 CUA revised guidelines for diagnosis and treatment of urological diseases.Color Doppler ultrasonography was used to check the prostate and to learn the prostate focal lesion,size, number and echo color Doppler flow characteristics. Of the 116 cases, 43 patients was undergone contrast-enhanced transrectal ultrasound. Results The biopsy results confirmed the diagnosis of prostate cancer was 64 cases, Benign prostatic hyperplasia was 52 cases. Of 43 cases who undergone contrast-enhanced transrectal ultrasound, Prostate cancer and Benign prostatic hyperplasia were 25 and 18 cases, respectively. CE-TRUS group and TRUS group showed no statistical difference between two groups. Analyzed the cases with PSA≤30 ng/ml, CE-TRUS group had a higher positive rate of biopsy (P=0.046). Conclusion TRUS guided transperineal biopsy of prostate might be an method for the diagnosis of prostate cancer with a higher accuracy rate. CE-TRUS can improve the biopsy positive rate of prostate cancer.

Objective To observe the anti-proliferation effects of rapamycin and paclitaxel of different hu-man prostate cancer cells in vitro. Methods The methods of MTr and flow cytometry were respectively ap-plied to observe the effect of rapamycin, paclitaxel and rapamycin+paclitaxel on proliferation and apoptosis of different prostate cancer cell lines (LNCaP-C4, LNCaP-C4-2, PC-3). Results When the concentration of rapamycin was 0.01 μmol/L, the impressive effect showed a remarkable difference in contrast to the con-trol. While in group LNCaP-C4-2 and PC-3, the goal concentration of rapamycin was 0.001μmol/L. When the concentration of paclitaxel was 0. 2 ng/mL, the impressive effect showed a remarkable difference in con-trast to the control. In group rapamycin (10 nmol/L) and in group paclitaxel (1 ng/mL) there were signifi-cant differences in growth inhibition, compared with control. While in group rapamycin(5 nmoL/L)+pacli-taxel(0.5ng/mL) there was significant difference in growth inhibition, compared with rapamycin (10 nmol/L) and paclitaxel (1 ng/mL) respectively. After cultured with rapamycin or paclitaxel alone, more tumor cells induced apoptosis than control. While after cultured with rapamycin and paclitaxel simultaneously, more tumor cells induced apoptosis than with rapamycin or paclitaxel alone. Conclusions Both rapamycin and paclitaxel had a good impressive effect on the three prostate cell lines (LNCaP-CA, LNCaP-C4-2, PC-3) with dose-dependent manner. After cultured with rapamycin and paclitaxel simultaneously, more tumor cells were induced apoptosis than with rapamycin or paclitaxel alone.

Objective To establish a method for rapid preparation of antiserum against influenza virus (H7N9) hemagglutinin,and to study the possibility of using it in single radial immunodiffusion (SRID) assay for quantitative detection of antigen in H7N9 influenza vaccine.Methods Hemagglutinin proteins expressed in eukaryotic cells were used to immunize sheep.Serum samples were collected to detect antibody titers by ELISA and double immunodiffusion assay.Different concentrations of antiserum were used in SRID assay to get the optimized concentration.Results After 4 times of immunization,the antiserum titers achieved 1 ∶ 1 000 000 and 1 ∶ 32 as indicated by ELISA and double immunodiffusion assay,respectively.The antiserum could form a clear precipitation line in SRID assay.The detection of antigen in the range of 10 to 40 μg/ml showed good linearity in the standard curve.The antigen titers in six batches of H7N9 vaccine detected by this SRID assay were identical with those by SDS-PAGE assay.Conclusion The antiserum against H7N9 hemagglutinin for SRID assay was developed successfully,and could be used as a reagent for the quantitative detection of antigen in H7N9 influenza vaccine.

Objective To compare the therapeutic effect of the combination of ~(99m)Tc-methnetium methylene diphosphunate and colloidal chromic,phosphate ~(32)p with the monotherapies in rats with adjuvant arthritis(AA).Methods In this study,50 Sprague-Dawley rats were randomly divided into five groups with 10 rats in each:normal control group(arthritis induced and treated with the same volume of normal saline via the same administration route),AA control group(arthritis induced with adjuvant and treated with normal sa- line),~(32)p colloid group(arthritis induced with adjuvant and treated with single intra-articular injection of col- luidal chromic phosphate ~(32)p 0.74 MBq and intra-peritoneal injection of normal saline every other clay),~(99m)Tc- MDP group(arthritis induced with adjuvant and treated with intra-peritoneal injection of ~(99m)Tc-MDP 2.5?10~(-3)?g/kg every other day and single intra-articular injection of normal saline),combination group(arthritis in- duced with adjuvant and treated with the combination of the two drugs).The diameter of the left hind ankle, serum levels of turnout necrosis factor(TNF)and interleukin-1?(IL-1?),and the pathological changes of an- kle joints were tested at different time points.Results The diameter of the left hind ankle at week 4 was smaller in the combination group than that of the ~(32)p colloid treatment alone group [(7.11?0.34)mm vs(7.57?0.29)mm,P

DNA, Viral ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Nucleic Acid Conformation ; RNA, Viral ; genetics ; Viral Envelope Proteins ; genetics

Objective To present the experience of laparoscopic extraperitoneal radical prostatectomy and evaluate its safety and efficacy. Methods A total of 91 patients diagnosed with localized prostate carcinoma were admitted from February 2003 to June 2008. The level of serum PSA ranged from 7. 5 - 47. 0 ng/ml(mean 14. 0 ng/ml). The volume of the prostate ranged from 35 - 75 ml(mean 52 ml). Biopsy was performed before the operation and the pathological results revealed prostate carcinoma with Gleason score no more than 8. CT, MR and ECT revealed there was no lymph node or seminal vesicle involvement and there was no bone metastasis. The procedures were performed with an-tegrade techniques and pelvic lymphadenectomies were performed in 32 cases and nerve-sparings were performed in 11 cases. Results The operation duration ranged from 105 - 270 min (mean 173 min). Intraoperative blood loss was 110 - 1200 ml(mean 315 ml). Incontinence occurred in 19 cases in early stage and 18 cases recovered within 3 months. Positive surgical margin occurred in 11 cases. There was no complication of urethra stricture during 3 - 30 months' follow-up. No lymph node was involved in 32 cases with pelvic lymphadectomy. Five of the 11 cases received nerve-sparing prostatectomy had normal erectile function during the follow-up. Conclusions Laparoscopic extraperitoneal radical prostatectomy is a safe, effective and efficient surgical procedure with the minimal invasion, less morbidity and rapid recovery. Laparoscopic radical prostatectomy is emerging as an alternative to open radical prostatectomy.

Background More efforts have been made in the functional protection of the glaucoma ganglion cells (RGCs) nowadays.As main ingredient,astragalus polysaccharides (APS) enhances neuron regeneration protein expression and promotes peripheral nerve recovery.But whether APS has a protecting effect on RGCs is incompletely clear.Objective The purpose of this study was to evaluate the neuroprotective effect of APS on the RGCs in a rat model of experimental glaucoma.Methods Forty-four SPF SD rats were divided into 4 groups randomly as follows:normal control group,negative control group,low dose APS group and high dose APS group,with 10 rats for each group.APS of 500 mg/kg or 2000 mg/kg (2.5 ml) was administered by gavage feeding once daily for 2 weeks in low dose or high dose of APS group,respectively,and the same volume of normal saline solution was applied instead of APS in the model control group.Two weeks later,aspirate 0.2 ml aqueous followed by methylcellulose injected into the anterior chamber to create the acute ocular hypertension model in the three groups above.No any intervention was performed in the normal control group.The rats were sacrificed on the fifth day after model established to take a retinal section.Ocular hypertension-induced damage was evaluated by regular retina histopathologic examination.Immunolhistochemistry for caspase-3 and TUNEL kits were used to determine the expression of caspase-3 protein in retina and apoptosis rate of RGCs.Retinal cross-sections were analyzed by Image Pro Plus 5.1 software to determine the thickness of various retinal layers and the positive staining cell density in the retinal ganglion cell layer (RGCL).Results On the fifth day after establishment of models,intraocular pressure (IOP) was significantly elevated in the model control group,low dose APS group and high dose APS group in comparison with the normal control group (t=-8.900,-10.700,-11.300,P＜0.01).Retinal morphology was normal in the rats of the normal control group,but in the model control group,rat retina was significantly thickened from severe retinal edema and cell arrangement disorder.Mild retinal abnormality was seen in the low dose APS group;while obvious retina edema was in high dose APS group.The entire retinal thickness,outer nuclear layer thickness and retinal nerve fiber thickness values were lower in the low dose APS group than those of model control group (t =-23.700,-14.770,-11.640,P＜0.01).However,no difference was found in outer nuclear layer thickness and retinal nerve fiber thickness values between high dose APS group and normal control group (t =-0.780,-0.460,P ＞ 0.05).Percentage of positive RGCs for caspase-3 protein and rate of apoptotic RGCs were significantly reduced in low dose APS group compared with model control group (caspase-3:F=87.710,P=0.001;RGCs apoptosis:F=272.840,P＜0.01).Conclusions 500 mg/kg APS can protect retina and RGCs against ocular hypertension-induced damage.The protection of APS is non-dosedependent.

Objective To summarize clinical experience of reconstructive operation with transpositional colon behind the sternum after corrosive esophageal burns and to explore the treatment for its complications.Methods Clinical data of 65 cases with esophageal scarred stricture after corrosive burns receiving reconstructive operation with transpositional colon behind the sternum were reviewed,56 of them by end-to-end anastomosis between transpositional anterograde peristaltic colon and esophagus,seven by end-to- end anastomosis between transpositional anterograde peristaltic colon and pharyngeal fundus,and two by end- to-end anastomosis between transpositional reversed peristaltic colon and esophagus,to summarize treatment experiences in pre-operation,operation and post-operation.Results Fifty-one of this group of patients recovered and discharged form the hospital smoothly,12 with cervical anastomotic leakage after operation including two cured by re-operation and ten cured by conservative treatment,and two with necrosis of transpositional colon including one died during operation and the other cured.Conclusions Corrosive burns of esophagus can be cured by leaving scarred stricture esophagus open without resection,and the effectiveness of reconstructive operation with transpositional colon behind the sternum is satisfactory with good pre-operative preparation,correct surgical operation,and correct post-operative treatment.

Objective To investigate the correlations of serum neuregulin 4 (NRG4) level with sex hormone and metabolism level in patients with polycystic ovary syndrome (PCOS). Methods A total of 87 PCOS patients were selected as the study subjects (PCOS group), and 90 healthy women (no reproductive system diseases, normal menstruation and normal sex hormone examination) were selected as the control group. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum NRG4 level, glucose and lipid metabolism indexes and sex hormone indexes. Pearson method and multiple linear regression were used to analyze the correlation between serum NRG4 and various indexes in PCOS patients. Results The serum NRG4 level in PCOS group was (24.41±7.31) ng/mL, which was significantly higher than (15.43±4.26) ng/mL in the control group (P < 0.001). Compared with the control group, the levels of BMI, hypersensitive C-reactive protein (hs-CRP), fasting blood glucose(FBG), fasting insulin (FINS), insulin resistance index (HOMA-IR), triglyceride (TG), luteinizing hormone (LH), luteinizing hormone/follicle stimulating hormone (LH/FSH), prolactin and total testosterone in PCOS group were significantly increased(P < 0.05). Serum NRG4 levels in PCOS patients were positively correlated with BMI, hs-CRP, FBG, FINS, HOMA-IR, TG, LH, LH/FSH and total testosterone (P < 0.05); the BMI, FBG and TG were the influencing factors of serum NRG4 level (P < 0.05). Conclusion The increase of serum NRG4 level in PCOS patients is associated with abnormal glucose and lipid metabolism and sex hormone indexes.