Objective To investigate the treatment effects and toxicities of extended-field intensity modulated radiation therapy (EF-IMRT) and intra-cavitary brachytherapy combined with chemotherapy for stage Ⅰ b1-Ⅳa cervical cancer with positive para-aortic lymph nodes.Methods A total of 46 stage Ⅰ b1-Ⅳa cervical cancer patients with positive para-aortic lymph nodes treated at Fudan University Shanghai Cancer Center between 2009 and 2011 were reviewed.Neoadjuvant,concomitant and adjuvant chemotherapy with paclitaxel and carboplatin were administrated for one cycle before radiation therapy,two cycles during radiation therapy or three cycles after radiation therapy.All patients received EF-IMRT and intra-cavitary brachytherapy.The positive lymph nodes received an additional boost dose.Results All patients received EF-IMRT to 50.4 Gy (1.8 Gy per fraction).Twenty-six patients was treated with boost dose of 6.0-8.0 Gy in 2.0 Gy per fraction to positive para-aortic lymph nodes.Thirty-seven patients received a positive paraaortic lymph nodes boost or(and) parametrial boost.All patient also received a high-dose-rate intra-cavitary brachytherapy at the point A dose of 20.0-30.0 Gy in 5.0 Gy per fraction.Total chemotherapy cycles were 189,and the average patient received 4.1 courses.Two cases (4％,2/46) experienced grade Ⅲ gastrointestinal toxicities,no patients suffered grade Ⅳ gastrointestinal toxicities.Fifteen cases (33％,15/46) experienced grade Ⅲ hematological toxicities,and 3 (7％,3/46) experienced grade Ⅳ hematological toxicities.Late grade Ⅲ-Ⅳ toxicity was seen in 3 cases (7％,3/46).The 3 year progressionfree survival rate was 46.2％,and the 3 years overall survival rate was 61.2％.Conclusion EF-IMRT and intra-cavitary brachytherapy combined with chemotherapy is safe and effective for stage Ⅰ b1-Ⅳa cervical cancer with positive para-aortic lymph nodes.

[Objective]To investigate the effects of mesenchymal stem cells (MSC) feeder layer, culture sere and freeze-thaw lysates on expansion and differentiation of cord blood CD34~+ cells in vitro. [Methods] MSC were isolated from human bone marrow and cultured until the third passage. Sera were obtained from the cultured MSC, and freeze-thaw lysates were obtained by repeated freeze-thaw procedures. Cord blood CD34~+ cells were isolated by magnetic cell separation system, and were co-cultured with the MSC feeder layer, culture sera, freeze-thaw lysates and hematopeietic growth factors (HGFs), respectively. The nucleated cells, CD34~+ cells, CD34~+CD38~- cells, CD41~+ cells and CD3~+ cells in the above culture system were detected by flow cytometry on day 6 and day 12. [Results] ①MSC feeder layer had a strong effect on nucleated cells, CD34~+,CD34~+CD38~- cells expansion. The MSC sera and freeze-thaw lysates had similar effect on cell expansion, but the effect was weaker than that of feeder layer (P<0.05). ② Both MSC sera and feeder layer inhibited cord blood CD34~+ cells differentiation toward CD3~+ cells or CD19~+ cells, and no significant differences were found between these two groups (P>0.05). ③ Both MSC sera and feeder layer promoted cord blood CD34~+ cells differentiation toward CD41~+ cells, and the effect was stronger in the feeder layer than that of the sera (P<0.05). ④ Freeze-thaw lysates had no effect on cell expansion and differentiation, and were similar with that of HGFs (P>0.05). [Conclusions] The MSC sera have positive effects on expansion of cord blood CD34~+ and CD34~+CD38~- cells, moreover they have the ability of promoting cord blood CD34~+ cells differentiation toward CD41~+ cells.

Objective:To observe autophagy induced by aminophylline in human peripheral blood T lymphocytes.Methods:Peripheral blood T lymphocytes from health adults were separated with Percoll(1.073 g/ml) and harvested by using nylon column.The cultured cells were divided into control group,aminophylline group(10-8～10-3mol/L,or 0.001 8～180 ?g/ml)and dexamethasone(DXM) group,and the rates of autophagy and apoptosis were analyzed by flow cytometry.Results:①The purities of T lymphocyte were 81.3%～94.5%。②In control group for 72 h cultivation,the autophagic rate of T lymphocytes was significantly lower than those of 10-5～10-3mol/L aminophylline group(P0.05);In 10-5mol/L aminophylline group,the autophagic rate of different T lymphocyte density after cultivation(0,24,48,72 h) had a tendency of increase along with time lasting and decrease of cell density,but the difference did not show significance.③There was significant statistic difference in incidence rate of apoptotic T lymphocytes between control and DXM group in 72 h cultivation(P=0.000);But there was no significant statistic difference in the rate of autophagic(P=0.481).Conclusion:10-3～10-5mol/L aminophylline can induce autophagy in peripheral blood T lymphocytes,10-3～10-8mol/L aminophylline could induce both autophagy and apoptosis,and there is no significant correlation between autophagy and apoptosis;DXM can induce apoptosis in peripheral blood T lymphocytes,which indicates different programmed cell death of T lymphocytes could be induced by different medicines.

Objective To explore the therapeutic effects of combined application of survivin antisense nucleic acid and taxol in subcutaneous xenograft mouse model of Balb/c and to preliminarily investigate the mechanism of the anticancer effects.Methods The model of subcutaneous tumor was established by hypodermic injection of C26 cells into Bal b/c mice.The mice were then randomly divided into five groups through the internal tumor injection:the blank group (C),lipo2000 group (L),paclitaxel group (T),survivin antisense nucleic acid group (A),and survivin antisense nucleic acid combined with paclitaxel group (A+T).We observed tumor growth,determined cell apoptosis by TUNEL method,and detected the expression of survivin by Western blot.Results ① All treatment groups had T/C<60%,which was significantly different from that of group L (P <0.05);the intervention was proved effective in vivo .The tumor inhibition rate of mice tumor weight showed that there were significantly curative effects in groups T,A and A+ T compared with that in group C (P < 0.05 ).The antitumor activity of paclitaxel (tumor inhibition rate of 21.82%±0.84%)could be improved by more than 59% through combination therapy (tumor inhibition rate of 54.1 6% ± 0.32%)concerning inhibition of tumor weight growth.② TUNEL method detected apoptotic cells:The tumor cells hardly had apoptosis in the blank group while T group and A group had a certain number of apoptotic cells.The experiment results suggested that PTX could promote tumor cell apoptosis,and that not only A+T strengthened the effect in killing tumor cells,but also the synergy of both could influence tumor resistance and ultimately make the effect in promoting tumor cell apoptosis conspicuous.③ The expression of survivin protein:The results showed that the expression of survivin protein in group A + T was obviously decreased without the expression of β-actin affected;it did not change significantly in group C compared with group L.The ratio of the A-value in groups T,A and A+T was 0.895 ±0.01 1,0.704 ±0.121 and 0.345 ± 0.01 9,respectively.Analysis of variance t-test showed that the expression level in group A+T obviously differed from that in groups C,L,A and T (P <0.05).Conclusion The combined therapy of survivin antisense nucleic acid and taxol can promote tumor cell apoptosis by downregulating the expression of survivin protein,reduce the body’s resistance to drugs and create synergetic effects.

Objective To evaluate the effect of the lactate clearance rate as a guide of the fluid resuscitation in multiple trauma patients with hemorrhage shock .Methods Sixty‐three multiple trauma patients with hemorrhage shock were divided into two groups ,which were group A (32 patients) and group B (31 patients) .In group A ,the central venous pressure (CVP) between8 and 12 mm Hg ,and the mean arterial pressure (MAP) (65 ± 5) mm Hg served as the goals of the fluid resuscitation .The B group ap‐plied the same goals of the CVP and MAP as group A .In addition ,the calculated lactate clearance rate (≥10% ) was used as the treatment target .The percent of patients reaching resuscitation goals and the ratio of the lactate clearance rate (≥10% ) at 2 hours after surgery ,the volume difference of infusion during the first 24 hours ,the incidences of organ dysfunction ,and the average days in ICU were compared between the two groups .Results The percent of patients reaching resuscitation goals at 2 hours after sur‐gery of group A was significantly higher than that of group B(P<0 .01) .The volume of the infusion of 24 hours in group A was less than that in group B(P< 0 .01) .There was no difference in the percent of patients reaching the lactate clearance rate at 24 hours after surgery(≥10% )and the infusin of red blood cells and plasma volume at 24 hours after surgery between the two groups (P>0 .05) .The incidence of organ dysfunction of group A was higher than that of group B (P<0 .05) .The average days in ICU of group A were longer than that of group B(P<0 .01) .Conclusion Based on monitoring CVP and MAP ,fluid resuscitation under the guidance of the lactate clearance rate(≥10% )may decrease the incidences of organ dysfunction and the days of patients in ICU .

Objective To investigate effect of mild hypothermia on changes of somatosensory evoked potential (SEP) and synaptophysin mRNA level after traumatic brain injury (TBI) and determine hypothermia-induced neuroprotection.Methods Forty-five SD rats were allocated into mild hypothermia group,TBI group and sham operation group with 15 rats per group according to the random number table.Left-side fluid percussion impact was performed to induce models of TBI.Rats were exposed to hypothermia environment (32-35℃) for 6 hours in mild hypothermia group after TBI.Rats in sham operation group were treated by only drilling on left side of the head,rather than hitting.To evaluate function outcome,modified neurological severity score (mNSS),SEP and synaptophysin mRNA level were measured at 6 hours,24 hours and 7 days postinjury.Results The mNSS in mild hypothermia group lowered compared with TBI group,especially at 24 hours and 7 days (P ＜ 0.05).SEP in mild hypothermia group was significantly shortened at 6 and 24 hours compared with TBI group (P ＜ 0.05),but SEP revealed no significant difference among the 3 groups at 7 days (P ＞ 0.05).Level of synaptophysin mRNA in mild hypothermia group increased at 6 hours postinjury compared with TBI group [(0.08 ± 0.02) vs (0.12 ±0.04)],with further increase at 7 days postinjury[(0.06 ± 0.01) vs (0.33 ± 0.10)] (P ＜0.05).Conclusion The shortage of nerve conduction time of the injured side and promotion of nerve regeneration suggest the neuroprotective role of mild hypothermia following TBI.

Objective To study the clinical outcomes of 28 pancreatic cancer patients who underwent total pancreaticoduodenectomy.Method The clinical data of 28 patients with pancreatic cancer who underwent total pancreaticoduodenectomy from January 2009 to March 2015 were retrospectively analyzed.Results Among the 28 patients,complications occurred in 11 (39.2％) after the operation.There were 7 patient having Grade Ⅱ,4 Grade] complications.No patient died within 30 days after the operation.Fol low-up of 25 patients showed a median survival of 13.5 months.There were 24 patients with pancreatic ductal adenocarcinoma,and the median survival was 13 months.Conclusions Total pancreaticoduodenectomy could not improve long-term survival but it decreased postoperative complications and improved postoperative quality of life.In selected patients,total pancreaticoduodenectomy could be a rational option.

Objective : To investigate the quality of life among people living with HIV/AIDS in Hangzhou City and analyze the influencing factors, so as to provide insights into the control of AIDS. Methods : From 1 January 2014 to 30 June 2018, the demographic characteristics, medical expenditures and disease status were collected from HIV/AIDS patients living in Hangzhou City, and the quality of life was assessed using the simplified Chinese version of Medical Outcomes Study-HIV Health Survey ( MOS-HIV ). Factors affecting the quality of life were identified among HIV/AIDS patients using multivariable linear regression analysis. Results : A total of 2 808 HIV/AIDS patients were surveyed, including 1 684 cases with HIV infections and 1 124 cases with AIDS. The participants included 2 510 men ( 89.39% ) and 298 women ( 10.61% ), and were predominantly at ages of 25 to 39 years ( 1 531 cases, 54.52% ). The physical and mental health scores were 53.87±6.96 and 46.03±9.09, respectively. Multivariable linear regression analysis identified age, average monthly income, self-paid medical expenses during the past year, and the latest CD4+T cell count as factors affecting physical and mental health ( P<0.05 ). Conclusions The quality of life is low among people living with HIV/AIDS in Hangzhou City, and is associated with age, income, medical expenditures and CD4+T cell count.

Objective To inquire into the location of the relevant gene mutations in the Chinese familial hypokalaemic periodic paralysis, and to specify the correlation between the genotype and the clinical features of this disease. Methods Target-exon PCR and DNA direct sequencing were used to research the mutations in the CACNA1S, SCN4A, and KCNE3 genes of 14 familial hypokalaemic periodic paralysis probands. If a positive member was found, the other members of his (her) family must be inspected with the sequencing method. Results The probands of 3 families showed the known correlating mutations of hypokalaemic periodic paralysis, which were R1239H mutations in the CACNA1S in 1 family and R672H mutations in the SCN4A in the other 2 families. In addition, the differences of the age of onset, the responsibility to the treatment with acetazolamide and penetrance were found between the CACNA1S R1239H and SCN4A R672H mutations. Conclusions SCN4A R672H and CACNA1S R1239H mutations exist in the Chinese familial hypokalaemic periodic paralysis. Differences of the clinical features exist, resulting from these 2 kinds of mutations.

Objective To investigate the phenotype and function of CD8+T cells cultured with SK-OV-3. Methods Transwell coculture experiments were conducted in 24-well plates with inner wells to separate CD8+ T cells and SK-OV-3. After 5 days of culture, CD8+ T cells were washed, and 1 × 106 cells were collected for Foxp3, CD25, CD28, CTLA-4 and GITR mRNA analysis and 2 × 106 cells were collected to detect expression of Foxp3, CD25, CD28, CTLA-4 and GITR in CD8+ T cells by flow cytometry. CD8+T cells that cultured alone or with SK-OV-3 were added at ratios of 1∶0 , 1∶1 , 1∶5 , 1∶10 , and 0∶1 to na?ve CD4+ T cells in 96-well plates. All wells were cultured with the presence of irradiated PBMCs and anti-CD3 antibody. After 72 h, [3H]-thymidine was added for 16 h prior to the determination of proliferation by scintillation counting. Results Compared with CD8+ T cells cultured without SK-OV-3 , the expression of Foxp3 and CTLA-4 was increased and CD28 expression was decreased in CD8+ T cells cultured with SK-OV-3 (both P < 0.001). We found that CD8+T cells cultured with SK-OV-3 significantly suppressed the na?ve CD4+ T cell proliferation induced by the anti-CD3 stimulation in a dose-dependent manner. In contrast, CD8+ T cells cultured without SK-OV-3 did not suppress na?ve CD4+ T cell proliferation. Conclusion Ovarian cancer cell can induce the suppressive CD8+Treg, which is an important link of the immunosuppressive microenvironment in ovarian cancer.