Objective To study the antitumor effects of photodynamic therapy (PDT)treated by ovarian cancer cell lysates in rat epithelial ovarian cancer in vivo. Methods Female Fischer344 rats of 6 -8 weeks were allocated to four groups ( n = 8 each) : PDT group ( inoculated intraperitoneal with PDT tumor cell lysates), freeze/thaw group ( inoculated intraperitonealy with freeze-thaw tumor cell lysates), normal saline group (inoculated intraperitoneal with normal saline) and control group. Rat epithelial ovarian cancer NuTu19 cells were injected into all rats by intraperitoneal at day 7,while injected with normal saline in control group. The number of tumor specific interferon-γ (IFN-γ) secreting splenocytes was quantified by enzyme linked immunospot(ELISPOT) assay, the cytotoxic T lymphocyte(CTL) activity of splenocytes was measured by lactate dehydrogenase(LDH) analysis and tumor growth and the survival time of rats were also observe& Results Stimulated by PDT tumor cell lysates, the number of tumor specific IFN-γ secreting splenocytes in PDT group, freeze/thaw group, normal saline group and control group were 448. 8±34. 2, 211.2±47.9,43.3 ± 11.1,16.1 ± 2.4 respectively, which were significant differences among of them ( P < 0.05). Stimulated by freeze/thaw tumor cell lysates, the number of tumor specific IFN-γ, secreting splenocytes in four groups were 151.7 ± 22.6,188.7 ± 53.0, 18.2 ± 12.2,8.8 ± 7.7 respectively, which were not significant differences among of them ( P>0.05 ). Cytotoxicity of splenocytes of PDT group increased significantly than that in other three groups(P <0.05). Except rats in control group were all alive until the experiment ended, the mean survival time of other rats were 234 d in PDT group, 171 d in freeze/ thaw group and 168 d in normal saline group, which in PDT group was significantly higher than those in freeze/thaw group and normal saline group ( P<0.05 ). Conclusions Rats treated by PDT tumor cell lysates could produce antitumor effects in vivo, which shown that induce tumor-specific immune response and prolong the life span.

Background Optic neuritis is closely associated with multiple sclerosis (MS).Its pathogenesis is uncompletely clear,and less basic researches are carried out at home and abroad. Objective This study was to reveal the expression of myelin basic protein (MBP) in the optic nerve of rat with experimental allergic encephalomyelitis (EAE) and to provide a theoretical evidence for the research of the relationship of optic neuritis with MS. Methods Fifty clean Wistar rats were randomized into the control group and immune 8,12,18 and 25 days groups.Myelencephalon was collected from 5 guinea pigs to prepare the homogenate and mixed with the isovolumetric complete Freud' s adjuvant (CFA).The 0.5 ml mixed antigen emulsifier was subcutancously injected into the 4 maps together with Bordetella pertussis 0.2 ml under the cutancous of dorsalis pedis at 0 and 48 hours to induce the EAE.Behavior of the rats was evaluated to score the neurological function.The optical nerve sections were prepared 8,12,18 and 25 days after immunology for the histopathological examination,and immunochemistry and Western blot were used to detect the expression of MBP in optic nerve.The use of the animals complied with the Regulation for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission. Results The disorder of motor nerve was seen 12 days following the immune,and the clinical neural functional scores were significantly higher 12 day and peaked on 18 days myelination and then gradually reduced.The histopathological examination showed that the irregular alignment of neural fiber was found at 12 days,and changes of cellular structure,edema of neural shaft bunch were observed at 18 days.However,the abnormal cells were significantly less 25 days following the immune.Immunochemistry showed that the MBP was expressed mainly in the myelination of optic nerve fibers.The numbers of positive cells for MBP were (115.75±26.49)cells/5 fields at the 12th day,showing a significant lowing in comparison with (167.44±22.49)cells/5 fields of control group (t=4.537,P＜0.05 ).The positive cells were lest at the 18th day with the values ( 75.57 ± 34.54) cells/5 fields ( t =6.362,P＜0.01 ).At the 25th day,positive cells increased to ( 117.63 ± 13.78) cells/5 fields,which still was lower than those of the control group ( t =4.068,P＜0.05 ).Western blot assay illuminated that with the prolong of immune time,MBP/β-actin ratio in optic nerve was gradually reduced and followed the same pattern at the 12th day( t =4.639,P＜0.05 ).At 18 days after immune in comparison with the control group,the expression of MBP/β-actin ratio in optic nerve was the least (t=8.427,P＜0.01). Conclusions MBP can be degraded in rat optic nerve.This is a further evidence that optic neuritis is a severe demyelination disease.It is clearly related to MS.

BACKGROUND:Transplantation of mesenchymal stem cells (MSCs) to multiple sclerosis (MS) is a new developed treatment to be given more and more attention.However,whether the MSCs function by cell replacement after going cross the blood-brain barricr or immune suppression needs further confirmation.OBJECTIVE:To establish a steady and effective method of Y chromosome in situ hybridization (ISH) and to detect the distribution of MSCs in a mouse model with MS.DESIGN,TIME AND SETTING:The observational study was performed at the Laboratory of Molecular Immunology,Institute of Healthy Science from September 2007 to February 2009.MATERIALS:A total of 30 C57BL/6 mice,aged 6-8 weeks and weighing 15-20 g,were selected.Female mice were used to establish experimental autoimmune encephalomyelitis (EAE).METHODS:Y chromosme specific DNA probe labled with digoxine (DIG) was designed and ISH was performed to confirm that the designed probe was Y chromosome specific and biologically sensitive.We transplanted the MSCs from male mice into the female EAE mice,and tracked the MSCs by Y chromosome ISH.MAIN OUTCOME MEASRUES:Distribution of MSCs in mice with EAE was observed under the optical and fluorescence microscope.RESULTS:The probe was confirmed to be Y chromosome specific and biologically sensitive.What's more,a steady and effective method of ISH was established.Some MSCs were detected in the periphery organs-spleen and lymph node of mice with EAE;seldom in spinal cord which indicated that MSCs might play its roles by immune suppression.CONCLUSION:Some MSCs were detected in the periphery organs-spleen and lymph node of mice with EAE,but seldom in spinal cord in the central nervous system.

Objective To establish chromatographic fingerprint of water and ethanol soluble component of Radix Dipsaci by using HPLC for identifying and evaluating its quality. Methods HPLC method was set up with Agilent HC-C18 column. The mobile phase was comprised of acetonitrile(A) and 0.05mol/L KH2PO4-H3PO4 buffer solution pH=3.0(B) in gradient elution mode. UV detection wavelength was at 220 nm,with a flow rate of 1.0 mL/min. Results Ten batches of Radix Dipsaci samples from different habitats and harvested in different times were analyzed. HPLC fingerprint of medicinal material of Radix Dipsaci was established preliminarily. Seventeen characteristic peaks were confirmed and the fingerprints of different samples were compared by similarity evaluation software. Conclusion This method shows high precision and good repeatability,so it can be used to assess the quality of Radix Dipsaci.

Objective To investigate the value of multi-slice spiral CT image texture analysis in differentiating metastatic (MLN) from non-metastatic lymph nodes (NLN) in patients with rectal cancer.Methods Thirty five patiets with rectal cancer who were pathologically confirmed by total mesorectum excision were included retrospectively,with regional lymph nodes (short-axis diameter of larger than 3 mm)found in preoperative CT images.All the patients underwent preoperative abdominal and pelvic dynamic contrast-enhanced CT scan.Regional lymph nodes were identified according to pathological findings,and were divided into MLN and NLN groups.The short-axis diameter,short-to long-axis diameter ratio of lymph nodes were manually measured and calculated,and the texture features,including skewness,kurtosis,variance,entropy and inverse difference moment,were analyzed.The above parameters between MLN and NLN groups were compared using independent sample t test or Mann-Whitney U test.ROC curve analysis was performed regarding the statistically significant parameters and the areas under curve (AUC) were calculated.Multivariate Logistic regression analysis was accomplished to obtain the independent predictive factor of diagnosing regional lymph nodes.Results A total of 68 regional lymph nodes were obtained and consisted of 31 MLNs and 37 NLNs.The short-axis diameter,kurtosis,and entropy of the MLN group were significantly higher than those of the NLN group (all P＜0.05).Whereas,the short-to long-axis diameter ratio,skewness,variance,and inverse difference moment did not differ significantly between the two groups (all P＞0.05).The AUC for distinguishing MLN from NLN of the short-axis diameter,kurtosis and entropy were 0.79,0.67,and 0.85,respectively.Multivariate logistic regression analysis showed that only entropy (odds ratio=8.48,95％ confidence interval was 3.01 to 23.92,P＜0.01) was screened out as the independent variable,which suggested that the entropy was the unique predictor for characterizing regional lymph nodes of rectal cancer.Conclusion Multi-slice spiral CT images texture analysis can facilitate the accurate differentiation between MLN and NLN in patients with rectal cancer,and especially the entropy has the optimal reference significance.

Objective To evaluate the effect of alveolar recruitment maneuver on the perioperative pulmonary function in the morbidly obese patients undergoing laparoscopic sleeve gastrectomy.Methods Forty morbidly obese patients of both sexes,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 18-64 yr,with body mass index ≥ 40 kg/m2,scheduled for elective laparoscopic sleeve gastrectomy,were randomly divided into either control group (group C) or alveolar recruitment maneuver group (group R) using a random number table,with 20 patients in each group.Patients in group C were treated with volume-or pressure-controlled ventilation after creation of pneumoperitoneum,maintaining the peak inspiratory pressure (Ppeak) ≤ 30 cmH2O and partial pressure of end-tidal CO2 35-40 mmHg.Patients in group R received alveolar recruitment maneuver once every 30 min starting from creation of pneumoperitoneum until the end of surgery.Patients were transfered to post-anesthesia care unit (PACU) with endotracheal tube which was extubated when the unified extubation standard was achieved in PACU.The patients who stayed in PACU for 2 h showing no indications for extubation were transfered to intensive care unit for continuous ventilation support.Immediately after intubation,immediately after creation of pneumoperitoneum,at 30,60 and 90 min of pneumoperitoneum,and at the end of pneumoperitoneum,blood samples were collected from the radial artery for blood gas analysis.Immediately after intubation,immediately after creation of pneumoperitoneum,at 30,60 and 90 min of pneumoperitoneum,at the end of surgery,and immediately before discharge from PACU,Ppeak,plateau pressure (Peat),and dynamic lung compliance were recorded.The time for achieving extubation standard and time for achieving the standard for discharge from PACU were recorded.Patients were followed up until discharge,and the feeding time and duration of hospital stay were recorded.Results Compared with group C,PaO2 and oxygenation index were significantly increased at 90 min of pneumoperitoneum,at the end of surgery,and immediately before discharge from PACU,Ppeak was decreased at 60 and 90 min of pneumoperitoneum and immediately after the end of pneumoperitoneum,Pplat was decreased at 60 and 90 min of pneumoperitoneum,the dynamic lung compliance was increased at 30,60 and 90 min of pneumoperitoneum and immediately after the end of pneumoperitoneum,and the time for achieving extubation standard,time for achieving the standard for discharge from PACU,feeding time,and duration of hospital stay were shortened in group R (P＜0.05 or 0.01).In group C,one patient did not present with indications for extubation and were transfered to intensive care unit for continuous ventilation support.Conclusion Intraoperative alveolar recruitment maneuver can effectively improve the intraoperative pulmonary function and promote the recovery of postoperative pulmonary function in the morbidly obese patients undergoing laparoscopic sleeve gastrectomy.

Objective To analyze the methylation status and protein expression of Ras association domain family- 1A (RASSF1A) in endometriosis (EMS). Methods The ectopic and corresponding eutopic endometrium tissues were collected from 45 women with EMS and normal endometrium tissues of 20 women without EMS. The methylation status of RASSF1A was examined by methylation specific PCR (MSP). Immunohistochemistry was performed to measure the level of RASSF1A in endometrium tissues. Results The RASSF1A protein expression rate in ectopic endometrium, eutopic endometrium, and normal endometrium was 37.78%(17/45), 60.00%(27/45) and 85.00%(17/20), and there was significant difference (χ2 = 13.136, P = 0.001). The frequency of aberrant methylation of RASSF1A was 55.56%(25/45), 33.33%(15/45) and 0 in ectopic endometrium , eutopic endometrium, and normal endometrium, and there was significant difference (χ2 =18.770, P = 0.000). The frequency of aberrant methylation of RASSF1A had no significant differnce throughout the menstrual cycle in ectopic endometrium and eutopic endometrium: 66.67%(14/21) vs. 45.83%(11/24), 38.10%(8/21) vs. 29.17%(7/24), P>0.05. In ectopic endometrium, the frequency of aberrant methylation of RASSF1A inⅢ-Ⅲstage was significantly higher than that in Ⅰ-Ⅱstage (χ2=5.940, P=0.015). In ectopic endometrium and eutopic endometrium, the RASSF1A protein expression had negative correlation with aberrant methylation of RASSF1A (r =- 0.594、- 0.577, P<0.01). Conclusions Epigenetic inactivation of RASSF1A through aberrant promoter methylation may be strongly correlated with the formation and progression of EMS, and assessment of RASSF1A methylation status in eutopic endometrium may be a potentially useful biomarker to enhance the early detection of EMS.

ObjectiveTo investigate the effect of recombinant human growth hormone (rhGH) on JAK2-STAT3 signaling pathway and on the growth of human colon cancer cells at different growth hormone receptor (GHR) expression status.MethodsLOVO and HCT-8 cell lines were selected after the colon cancer cells were screened using flow cytometry according to the GHR expression status.The growth of human colon cancer cells after intervention with rhGH was detected by MTT assay.The proliferation index ( PI),cell cycle,and apoptosis were detected by flow cytometry.The expression of JAK2-STAT3 signaling pathway proteins was detected by Western blot.ResultsHCT-8 cell line showed positive GHR expression (59.6％),and LOVO cell showed negative GHR expression (3.5％).The growth rate of HCT-8 cells increased after rhGH treatment.Compared with the untreated HCT-8 cells,the rhGH-treated HCT-8 cells showed reduced apoptosis,elevated PI,and increased G2/Mphase cells ( P =0.0073).Western blot revealed that rhGH up-regulated the proteins of pJAK2,pSTAT3,VEGF,Cyclin D1,and Bcl-xL in HCT-8 cells.In contrast,no such changes were observed in LOVO cells treated with rhGH.ConclusionsrhGH may promote the growth of HCT-8,the cell line of high GHR expression,and up-regulate the expression of a number of key nodes in JAK2-STAT3 signaling pathway.However,rhGH may not affect LOVO,the cell line of low GHR expression.

Objective To construct tissue engineering nucleus pulposus by culture of rabbit bone marrow mesenchymal stem cells (rBMSCs)-nucleus pulposus acellular matrix scaffold (NPAMS)complexes (rBMSCs-NPAMS).Methods Several NPAMS were prepared,and rBMSCs was seeded into NPAMS.The scaffolds and complex were detected by general observation,HE stai-ning,immunohistochemical,qRT-PCR,scanning electron microscopy.Results The scanning electron microscopy showed the seed cell in nucleus pulposus ECM-derived scaffold could adhesion and growth.The cell attachment and proliferation were observed by HE staining.Immunohistochemical examination with typeⅡ collagen showed positive results.qRT-PCR revealed the time-depend-ent of the mRNA expression of collagen Ⅱ,and which was smaller than positive control(P<0.01).The relative expression of Ag-grecan mRNA grew in a time dependent fashion,which was still lower than positive control(P<0.01).Scaffolds and normal nucle-us pulposus had no statistical significance of the compression load under the same displacement of the compression(P>0.05).Con-clusion Natural nucleus pulposus acellular matrix scaffold composite allogeneic bone marrow mesenchymal stem cells can be suc-cessfully built into tissue engineering nucleus pulposus.

Objective To observe the clinical effects and safety of radiofrequency thermocoagulation in treating the lumbar intervertebral disc protrusion.Methods Thirty patients with lumbar intervertebral disc protrusion,whose diagnoses were confirmed by clinical manifestations and CT findings,were involved in this study.The needle was punctured to the target point of the diseased intervertebral space under C-arm fluoroscopic guidance.After the testing of sensory nerve,motor nerve and temperature was made,the target needles were heated until the nerve radiofrequency temperature meter reached the point of 92℃.This point of temperature was held for 100 seconds and the procedure was repeated for four cycles.Results Six months after the treatment,all patients showed an obvious improvement in VAS,which decreased from 7.83±0.33before operation to 2.37±0.48 after treatment(P＜0.05).According to the modified Macnab therapeutic evaluation criteria,excellent result was seen in 9,good result in 12,fair result in 5 and poor result in 4cases,with a total effective rate of 86.7%.No serious complications occurred in all patients.Conclusion Radiofrequency thermocoagulation is an effective and safe method for the treatment of lumbar intervertebral disc protrusion.