Objective To investigate the blood-saving effect of controlled low central venous pressure (CLCVP) in different types of hepatectomy.Methods Ninety ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 37-76 yr,weighing 40-75 kg,undergoing elective hepatectomy,were divided into 6 groups according to the surgical approach and whether CLCVP was used during surgery (n =15 each):CLCVP1-3 groups and nonCLCVP1-3 groups (NCLCVP1-3 groups).The standard hepatectomy,half liver resection and irregular hepatectomy were performed in CLCVP1-3 groups,respectively,with CLCVP.The standard hepatectomy,half liver resection and irregular hepatectomy were performed in NCLCVP1-3 groups,respectively,without CLCVP.In CLCVP1-3 groups,from skin incision to the end of liver resection,CVP was maintained ≤ 5 cm H2 O through adjustment of the position,fluid restriction and iv infusion of nitroglycerin,and norepinephrine was infused simultaneously to maintain mean arterial pressure ≥ 60 mm Hg.In NCLCVP1-3 groups CVP was maintained at 6-12 cm H2O.Intraoperative blood loss and blood transfusion were recorded.Results Compared with NCLCVP1-3 groups,intraoperative blood loss was significantly decreased in CLCVP1-3 groups (P ＜ 0.05).Compared with NCLCVP3 group,the amount of blood transfusion was significantly decreased,the constituent ratio of intraoperative blood loss ＜ 200 ml was increased,and the constituent ratio of intraoperative blood loss ＞ 1000 ml was decreased in group CLCVP3 (P ＜ 0.05).Conclusion CLCVP can decrease the intraoperative blood loss and blood transfusion in patients undergoing irregular hepatectomy.

Objective Design and synthesize short hairpin RNA (shRNA) expression vector of RNA for specific silencing of heparanase (HPA) gene,screened plasmid which silence effects is the best.Observe the function of ceil invasion after inhibiting the expression of HPA in cervical carcinoma cell lines (HeLa).Methods The genomic sequence of HPA gene was retrieved from GenBank database.Designed four pairs of specific oligonucleotide sequences and a negative control according to the shRNA design principles.They were inserted into the vector pYr-1.1,vectors,and transfected into HeLa cells via lipofectamine.Reverse transcription (RT)-PCR and immunofluorescence were employed to detect the expression of HPA gene in the transfected cells at the mRNA and protein levels,respectively.The plasmid were screened and transfected into HeLa cells,then transwell small room stromal invasion experiment were employed to observe the cervical carcinoma cell invasion.Results RT-PCR results of transfected HeLa cells shown that the mRNA amplification multiples were 0.54 ±0.05 in the HPA-592 group,0.89 ±0.18 in HPA-995 group,0.82 ±0.22 in the HPA-1351 group,0.91 ±0.47 in HPA-1658 group.While,they were 1.31 ±0.72 and 1.09 ±0.16 in negative control and blank control group,respectively.Green fluorescence was visible in the cytoplasm,which indicated that the HPA protein was expressed in the cytoplasm,of them the weakest green fluorescence in the HPA-592 group.The relative numbers of invasive cells among the HeLa cells were as follows:182 ±6 in the blank control group,258 ± 17 in the negative control group,and 44 ± 4 in the HPA-592-specific interference group(P ＜ 0.01).Conclusion Successfully screened shRNA vector targeting human HPA,efficiently inhibit expression of HPA gene when transfected into HeLa cells,and significantly reduced the invasion capacity of cervical carcinoma cells.

Objective To detect the expression of heparanase (HPA) in cervical cancer cells and investigate the impact of quercetin on the expression of HPA,and the molecular mechanism that quercetin inhibits the growth of cervical cancer cells.Methods The experimental groups included cervical cancer cell lines (HeLa and Caski) exposed to different concentrations of quercetin (20,40 and 80 μmol/L) in the culture medium.The control groups included a negative control group,which was cultured with RPMI 1640 only,and a positive control group,in which cervical cancer cells were transfected with HPA small interference RNA (siRNA) to silence HPA expression.The cellular expression levels of HPA were detected with fluorescence quantitative real-time PCR and western blot analysis at 24,48 and 72 hours after treatment.Results (1) HPA was significantly expressed in both cervical cancer cell lines (HeLa and Caski),and it exists both nucleus and cytoplasm.(2)The real-time PCR shows as follows:as the quercetin concentration increased (20,40 and 80 μmol/L),the mRNA expression level of HPA decreased (P ＜0.01),in which the inhibition of HPA expression was concentration dependent.In addition,the inhibition of HPA expression was also time dependent.As time growth,the expression level of HPA mRNA (24,48 and 72 hours) in HeLa and Caski cells decreased (P ＜ 0.01).Compared with negative control group,the expression level of HPA mRNA decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ; Compared with positive control group,the expression level of HPA mRNA expressed no obvious difference in quercetin (80 μmol/L) group (P ＞ 0.05) in HeLa cells,while it was opposite in Caski cells(P ＜0.01).(3)The result of western blot shown that,as the quercetin concentration increased(20,40 and 80 μmol/L)and time growth (24,48 and 72 hours),the expression level of HPA protein decreased (P ＜ 0.01),and the inhibition of HPA protein expression was concentration and time dependent.Compared with negative control group,the expression level of HPA protein decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ;Conpared with positive control group,the expression level of HPA protein expressed no obvious difference in quercetin (80 μmol/L) group (all P ＞ 0.05) in both HeLa cells and Caski cells (all P＞0.05).Conclusion Quercetin could inhibits the expression of HPA in cervical carcinoma cell lines,which inhibition is concentration and time dependent.

Objective:To observe the clinical efficacy of Ba-pulling and Qian-traction manipulation with neck suspension and movement for acute cervical radiculopathy. Methods: A total of 85 patients who met the inclusion criteria were randomized into an observation group and a control group by random numbers, with 43 cases in the observation group and 42 cases in the control group. The observation group was treated with Ba-pulling and Qian-traction manipulation with neck suspension and movement;while the control group was treated with Bashen-pulling and stretching manipulation in a supine position. The treatment was performed once a day, 10 times as a treatment course. The therapeutic efficacy was evaluated after 1 treatment course, and the changes in the scores of visual analog scale (VAS) and neck disability index (NDI) were observed. Results: The total effective rate was 97.7% in the observation group, and 83.3% in the control group, and the difference between the two groups was statistically significant (P＜0.05). After treatment, the VAS and NDI scores of both groups were significantly decreased (both P＜0.01), and the differences in the VAS and NDI scores between the two groups were statistically significant (both P＜0.01). Conclusion: Both Ba-pulling and Qian-traction manipulation with neck suspension and movement and Bashen-pulling and stretching manipulation in a supine position can relieve pain and improve cervical function in patients with acute cervical radiculopathy, and Ba-pulling and Qian-traction manipulation with neck suspension and movement can produce more significant efficacy than Bashen-pulling and stretching manipulation in a supine position.

Objective To evaluate the radical effect and prognosis of laparoscopic resection for colorectal cancer in China.Methods Articles of non.randomized comparative studies(NRCs)of laparoscopic resection and open Burgery for colorectal cancer which were published before October 2007 were retrieved,and correlated indexeswere extraeted for meta.analysis.Results The mean quality score of the 14 articles selected was 18.92±1.27.The basic characteristics of patients in laparoscopic resection group were similar to those in open surgery group.Compared with open surgery group,the incised length of the intestine in the laparoscopic resection group was shorter by 0.66 cm.and the distance between distal margin of resection and tumor was farther by 0.26 cm.The 2-year survival rate of patients in laparoscopic group Was 1.67 times higher than that of open surgery group.There was no significant difference upon follow-up rate,tumor diameter,number of resected lymph nodes,local recurrence rate and distal metastasis rate between the 2 groups.Conclusions The results of meta-analysis show that laparoscopic resection has the sanle radical effect as open surgery for colorectal cancer.but the 2-year survival rate of patients treated by laparoscopic resection is comparatively higher.

Objective To study the efficacy and safety of Gonadotropin-releasing hormone agonists (GnRH-a) combined with laparoscope conservative surgery in treatment of moderate or severe endometriosis.Methods From Jan.2007 to Jan.2010,68 patients with moderate or severe undergoing treatment in Renmin Hospital of Wuhan University were enrolled in this retrospective study.Three groups were classified,which were 25 patients in GnRH-a group,subcutaneous injection Leuprorelin on the second day of menstruation,every 4 weeks for 3 months.Twenty-three patients in Marvelon group,orally one marvelon tablet on the second day of menstruation,continuous 21 days for one period of treatment for 3 courses.Twenty patients in surgery group,without any medicine used preoperatively.All patients were followed by 12 months and compare their surgery time,blood loss,recovery,visual analog scale (VAS),and recurrence and so on.Results The operating time were (68 ± 18) min in GnRH-a group,(80 ± 21) min in Marvelon group and (90± 24) min in surgery group.The amount of bleeding were (118 ± 15) ml in GnRh-a group,(161 ± 18) ml in Marvelon group and (193 ± 13) ml in surgery group.There was significant lower in the operating time and amount of bleeding in GnRH-a group than those in other two groups (P ＜ 0.05).The activity time and the anus exhaust time were shorter in patients in GnRh-a group than those in the other two groups significantly(P ＜ 0.05).When followed up in 12 months after treatment,visual analogue scale had dropped from 3.8 (1.9-6.8) to 1.9 (1.1-2.8) in GnRh-a group,from 2.7 (1.3-5.5) to 1.8 (1.2-3.2) in Marvelon group and from 1.9(1.0-4.9) to 1.6(1.0-3.6) in surgery group.It was showed the most remarkable decreased VAS in GnRHa group when compared with the other two groups(P ＜ 0.05).The recurrence rates were 12％ (3/25) in GnRH-a group,22％ (5/23) in Marvelon group and 25 ％ (5/25) in surgery group.It was found that the most significant lower recurrence was in GnRH-a group when compared with the other two groups (P ＜ 0.05).Conclusions It was safe and efficacy that GnRH-a combined with laparoscopic conservative surgery were used in treatment of endometriosis.It could bring shorter operation time,less intraoperative blood loss,quick postoperative recover,the lower recurrence rate.

OBJECTIVE: To analyze the indications of the therapies for rheumatoid arthritis (RA) with neural network model analysis. METHODS: Three hundred and ninety-seven patients were included in the clinical trial from 9 clinical centers. They were randomly divided into Western medicine (WM) treated group, 194 cases; and traditional Chinese herbal medicine (CM) treated group, 203 cases. A complete physical examination and 18 common clinical manifestations were prepared before the randomization and after the treatment. The WM therapy included voltaren extended action tablet, methotrexate and sulfasalazine. The CM therapy included Glucosidorum Tripterygii Totorum Tablet and syndrome differentiation treatment. The American College of Rheumatology 20 (ACR20) was taken as efficacy evaluation. All data were analyzed on SAS 8.2 statistical package. The relationships between each variable and efficacy were analyzed, and the variables with P<0.2 were included for the data mining analysis with neural network model. All data were classified into training set (75%) and verification set (25%) for further verification on the data-mining model. RESULTS: Eighteen variables in CM and 24 variables in WM were included in the data-mining model. In CM, morning stiffness, swollen joint number, peripheral immunoglobulin M (IgM) level, tenderness joint number, tenderness, rheumatoid factor (RF), C-reactive protein (CRP) and joint pain were positively related to the efficacy, and disease duration and more urination at night negatively related to the efficacy. In WM, erythrocyte sedimentation rate (ESR), weak waist, white fur in tongue, joint pain, joint stiffness and swollen joint were positively related to the efficacy, and yellow fur in tongue, red tongue, white blood negatively related to the efficacy. In the analysis with the neural network model in the patients of verification set, the predictive response rates of 20% patients would be 100% and 90% in the treatment with CM and WM, respectively. CONCLUSION: Neural network model analysis, based on the full clinical trial data with collection of both traditional Chinese medicine and modern medicine diagnostic information, shows a good predictive role for the information in the efficacy in rheumatoid arthritis.

OBJECTIVETo study the specific cytotoxicity of (131)I-Rituximab against CD20-positive B-cell lymphoma cells.

METHODSRituximab was labeled with (131)I using IODO-GEN method, and the dose-effects of various concentrations of (131)I-Rituximab, (131)I alone and Rituximab in Raji cells were evaluated by MTT assay to determine the optimal dose according to the dose-effect curves. The cytotoxicity of (131)I-Rituximab, (131)I and Rituximab was assessed in CD20-positive Ramos (RA-1) cells, Raji cells and CD20-negative Molt-4 cells according to the changes of the survival rates. Giemsa staining was used to evaluate the antitumor effect of (131)I-Rituximab in Raji cells by measuring the mitosis index (MI).

RESULTS(131)I-Rituximab presented with a dose-dependent cytotoxicity against Raji cells. At the specific activity of 60 microCi/ml, (131)I-Rituximab resulted in significantly higher growth inhibition rate of the cells than Rituximab (P<0.05). The inhibition rate of Raji cells treated with (131)I-Rituximab, (131)I, or Rituximab for 96 h were comparable with the rates in Ramos RA-1 cells, but significantly higher than the rates in Molt-4 cells. The MI values in (131)I-Rituximab group were significantly lower than those in the other groups (P<0.001).

CONCLUSION(131)I-Rituximab can induce specific cytotoxicity against CD20-positive tumor cells, and may potentially serve as an agent for targeted radioimmunotherapy for CD20-positive B-cell lymphoma.

Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Murine-Derived ; Antigens, CD20 ; immunology ; Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Iodine Radioisotopes ; pharmacology ; Lymphoma, B-Cell ; pathology ; Radioimmunotherapy ; Tumor Cells, Cultured

Objective:To analyze the protective effects of adoptively transferring different patrilineal lymphocytes and their Exosomes ( Exo) on fetation of mice with pregnancy loss comparatively.Methods: The peripheral blood mononuclear cells ( PBMC) from healthy men and the splenocytes from BALB/c and DBA/2 male mice were induced in vitro ,and their Exo were isolated through sucrose gradient ultra-centrifugation combined with ultrafiltration.The mice of CBA/J (♀) mated with BALB/c (♂) were enrolled as control group of normal pregnancy ,and the CBA/J (♀) mated with DBA/2 (♂) as URSA of pregnancy loss experimental animal model.The mice in URSA group were randomly divided into each group with treatment through adoptively transferring , which were injected intravenously or subcutaneously with splenocytes or splenocytes -derived Exo from mated DBA/2,unmated DBA/2 or unrelated BALB/c,also PBMC-derived Exo from men,respectively.And then,the placenta volumes,rates of fetal absorption and pregnancy loss were calculated to observe the fetation of embryos.Results:Compared with the group of normal pregnancy ,the placenta volumes from URSA group decreased greatly ,and rates of fetal absorption and pregnancy loss elevated greatly ( all P<0.000 5 ).After transferring different sources of cells and their Exo through different injection ,the placenta volumes resumed to the level of normal pregnancy ,and the rates of fetal absorption and pregnancy loss decreased significantly ( all P<0.000 5 ).No differences were observed after treatment through injecting intravenously or subcutaneously ( all P>0.05 ).After transferring the Exo derived from either male mice or healthy men,the level of decreased fetal absorption rates were more than that in cellular-therapy groups ( all P<0.05 ).After transferring the Exo derived from men ,the level of decreased pregnancy loss rates were more than that in cellular -therapy groups and mice splenocytes-derived Exo group ( all P<0.05 ).Conclusion:Adoptively transferring patrilineal T lymphocytes and their Exo can greatly improve the fetation.Exo should become a non-cellular bio-remedy,which is expected to replace traditional immunotherapy of adoptively transferring lymphocytes.

OBJECTIVETo observe the killing effect of Herceptin and adriamycin sequentially applied on breast cancer cell line in vitro.

METHODSBT-474 human breast cancer cells in exponential growth phase were treated with Herceptin alone, adriamycin alone and their sequential administration (Herceptin before adriamycin and vice versa), respectively. Under optical microscope, the morphological changes of the cells were observed before and after drug administration. The expression rate and mean fluorescence intensity (MFI) of HER-2/neu and cell death rate were detected by flow cytometry.

RESULTSMicroscopically, the cells treated with different protocols all exhibited such changes as darkening and increase of cellular debris with irregular cell morphology. Flow cytometry revealed no significant difference in the expression rate of HER-2/neu in each group before and after treatment, but the MFI of HER-2/neu and death rate of the treated cells were significant different from those of the control group (P<0.05). The cell death rate of Herceptin-pretreated cells was significantly higher than that of adriamycin-pretreated ones (P<0.05).

CONCLUSIONHerceptin pretreatment enhances the killing effect of adriamycin on breast cancer cell line BT-474, which provides experimental evidence for designing clinical sequential biochemotherapy of breast cancer.

Antibiotics, Antineoplastic ; pharmacology ; Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Death ; drug effects ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Synergism ; Female ; Flow Cytometry ; Humans ; Receptor, ErbB-2 ; biosynthesis ; Trastuzumab