Objective To study the impact of different low concentrations of cerium oxide for hepatocellular carcinoma cell prolifera-tion.Methods Three different types of hepatoma cells (Huh7, HepG2,7721) were cultured,and added different concentrations of cerium oxide (0.005,0.01,0.05,0.1,1 μg/mL),of which the cell proliferation was detected by CCK8.The apoptosis-related genes was detected by qRT-PCR technology.The cell cycle was analyzed by flow cytometry.And the effect of low concentration cerium oxide on hepatocellular carci-noma cells tumorigenicity was confirmed by the nude mice experiments.Results CCK8 experiment showed that low concentrations of cerium oxide could promote proliferation of hepatocellular carcinoma cell, especially in concentration of 0.01μg/mL.The qRT-PCR showed that low concentration of cerium oxide could inhibit the apoptosis of hepatocellular carcinoma cell.The flow cytometry analysis had not found any effect of cerium oxide on cell cycle.The tumorigenicity experiments confirmed that low concentrations of cerium oxide could enhance the tumorigenic ability of hepatocellular carcinoma cell.Conclusion Low concentration of cerium oxide can significantly improve the proliferation of liver cancer cells.

Objective To amplify natural killer (NK) cells in vitro and explore its killing effect on ovarian cancer cells.Methods (1) The separation of NK cells and identification.A total of 20 ml peripheral blood of one healthy volunteer was collected in Nov.2015,Peking University People's Hospital.The peripheral blood mononuclear cells of normal volunteers were isolated,cultured in vitro and amplificated cultivation for 14 days with K562 cells transfected and expressing interleukin 21 (IL-21-K562) as nourish cells.The number and dynamic state of the growth cells were monitored during the cultured process.Cells were harvested and counted after 14 days cultured.The NK cells phenotypes were detected by flow cytometry.(2) The killing effect of NK cells on ovarian cancer cells:the ratio of effector cells (NK cells) and target cells (ovarian cancer cells and its control) was 50∶ 1,20∶ 1,10∶ 1,5∶1 or 1 ∶ 1,NK cells killing effect on ovarian cancer cells was detected by the lactate dehydrogenase (LDH) release experiments.Results (1) The results of NK cells establishment and phenotypic characterization:the cells were induced in vitro for 14 days by amplification culture.With the extension of incubation time,the number of NK cells increased constantly,from 2.0× 107 on day 0 to 5.1 × 109 on day 14.Obvious amplification of the total number of cells were detected for 255 times.Living cells unstained by trypan blue eventually reached 95％ above.Before and after the induction and amplification in vitro,the percentage of NK cells (CD3-CD56+cells) in CD3-cells were 2.33％ and 85.32％,respectively (P＜0.01),which covered the whole lymphocytes 1.06％ and 69.42％,respectively (P＜0.01),which showed that NK was the main cell type in the amplificated lymphocytes.(2) The killing rate of NK cells on ovarian cancer cells in vitro:the results detected by LDH release experiments showed that NK cells could performed strong nonspecific killing effect on ovarian cancer cell lines SKOV3,HOC1A,3AO and CAOV3,as well the normal ovarian cell line T29 and NK sensitive cell line K562,and the killing effect increased significantly along with the increase of effector cells and target cells ratio (P＜0.01).When the ratio was 1 ∶ 1,the killing rate was 37％ for K562,while the rate of killing of other cells was around 10％ (P＜0.05).When the effect-target ratio was 20∶1 and 50∶ 1,in addition to CAOV3 cells (more than 70％),NK cells had a kill rate of more than 80％ for other ovarian cancer cells lines and their control cell K562 and T29 cells (P＞0.05).Conclusion NK cells could be established in vitro and have a good non-specific killing effect on ovarian cancer cells.

This paper introduces the background, implication and construction of the county health services community (County medical alliance) model in Anhui Province under the background of new medical reform,and briefly introduces the relationship between medical insurance, enhancing the ability to upgrade and standardize services and medical treatment integration of the typical experience.It also analyses the challenges faced in the construction of medical syndicate, such as the mechanism of regional environmental restriction, compensation and assessment mechanisms which have not been established yet, and the sustainable development of information technology that has lagged behind, and put forward the policy suggestion to improve the construction of medical community, with a view to providing reference for the next work.

Objective To investigate the role of TRPV1 in exacerbation of gastric mucosal injury in a rat water immersion restraint stress (WIRS) model by acute postoperative pain. Methods Thirty Wistar rats were randomly divided into normal controlled group (N group, n=10), WIRS model group (WIRS group, n=10) and surgery after WIRS group (WS group, n = 10). The general extent of gastric mucosal injury was observed and assessed for gastric mucosal ulcer index (UI), intragastric pH and serum SOD/MDA ratio were measured and the expression of TRPV1 mRNA in gastric mocusal was accessed by Real-time Quantitative PCR. Immunohistochemistry was performed to detect the mean density of TRPV1. Results Compared with NC group, WIRS group showed obvious gastric mocusal injure with higher UI , lower values of intragastric pH serum SOD/MDA ratio and TRPV1 (P<0.05). The treatment with surgery after onset of WIRS significantly aggravated the gastric mucusal erosion and hemorrhage, with UI increased (P < 0.05), the value of intragastric pH, serum SOD/MDA ratio and TRPV1 further reduced (P < 0.05). Meanwhile, TRPV1 was inversely correlated with UI, and positively associated with intragastric pH and serum SOD/MDA ratio. Conclusion TRPV1 expression in gastric mocusal of AMGL model is inhibited by acute postoperative pain. TRPV1 may involve in the exacerbation of gastric mucosal injury in WIRS model by acute postoperative pain.

Objective To identify the T cell epitopes from ovarian cancer associated anti-idiotypic antibody 6B11in order to explore the molecular basis of 6B11 induced cellular immune responses against ovarian cancer.Methods Potential human leukocyte antigen(HLA)A0201 ligands were predicted by using SYFPEITHI algorithm and tested by the T2 binding assay for screening of HLA-A2 binding peptides from 6B11 complimentary determining region(CDR).Cytotoxic T lymphocytes(CTL)to 6B11 or peptides were generated by 3 rounds of in vitro stimulation with 6B11 or peptide-pulsod dendritic cells(DC),and then tested by 51Cr-release assay to ascertain the CTL epitope of 6B11.Cell proliferation assay was performed by using 6B11 specific CTL as responder cells and peptide-pulsed DC as stimulators to ascertain the helper T lymphocyte(Th)epitope of 6B11.Cytokine assay and interferon-γ ELISPOT assay were performed to verify the CTL and Th epitope of 6B11 further.Results Light chain CDR3 peptide(VL CDR3)of 6B11 induced specific CTL to kill HLA-A2 and target antigen positive ovarian cancer cells,which could be blocked by anti human major histocompatibility complex(MHC)Ⅰ antibody.Heavy chain CDR3 peptide(VH CDR3)of 6B11 stimulated the proliferation of 6B11-primed CTL,which could be blocked mainly by anti human MHC-Ⅱ antibody,and further experiments showed that part of the VH CDR3 peptide-primed CTL killed K562 cells.Peptides in VL CDR3 and VH CDR3 were the CTL and Th epitope mimicking the original antigen of 6B11 respectively.Collaboration of 6B11 CTL and Th epitope,or 6B11 CTL epitope and keyhole limpet hemocyanin(KLH),the former was more powerful in inducing specific cellular immune responses against ovarian cancer.6B11 or corresponding CTL and Th epitope specific CIL secreted high levels of interleukin-2 (1630,1503 ng/L)and interferon-γ(5620,5421 ng/L),while basal level of interleukin-4 was detected (253,274 ng/L).ELISPOT assay confirmed the existence of specific interferon-γ secreting cells in 6811 or epitopes specific CTL(196/1×106 T cell,184/1×106 T cell).Conclusions VL CDR3 and VH CDR3 peptides of 6B11 are the CTL and Th epitopes mimicking original antigen which could duplicate the activity of intact 6B11 to induce the cellular immune responses against ovarian cancer.The results have significant theoretical and practical value in application of anti-idiotypic antibody ag anti tumor vaccine.The acquired CTL and Th epitopes as constituents of future multiple peptides against ovarian cancer could be used in peptide vaccine based ovarian cancer therapy.

Objective To investigate the value of human epididymis secretory protein 4 ( HE4 ) combined with CA125 assay in differential diagnosis of endometriosis cyst and ovarian malignant tumor.Methods The level of HE4 and CA125 were measured by enzyme-linked immunosorbent assay (ELISA) in the serum specimens of 46 cases in endometriosis cyst group,36 cases in malignant ovarian tumor group,60 cases in benign ovarian diseases and 50 women in healthy women group.Those results were shown with median level.The normal range were 0-150 pmol/L in HE4 and 0-35 kU/L,which either one was more than the threshold value defined as positive index.The sensitivity of assay was evaluated by receiver operating characteristic (ROC) curve,the relation and value of HE4 or CA125 alone and combination assay in diagnosis of endometriosis was analyzed by Mann-Whitney U test and correlation analysis.Results (1) HE4:the median levels of HE4 were 52.4,51.0,50.0 pmol/L in group of endometriosis,normal control and benign ovarian tumor,which didn't show statistical difference.However,HE4 was 507.5 pmol/L inovarian cancer group,which was significantly higher than those of 3 groups (P < 0.05 ).(2 ) CA125:there were significant different in median level of CA125 was observed as 743.0 kU/L in ovarian cancer,84.9 kU/L in endoemtriosis,15.4 kU/L in benign ovarian disease,and 11.5 kU/L in healthy women (P < 0.05).( 3 ) The single assay:when compared with that in endometriosis group,receiver operating characteristic area under the curve( ROC-AUC) were 0.933 in HE4 alone and 0.821 in CA125 alone assay in ovarian cancer group.The specificity was 95% and the sensitivity was 79.6% and 49.0%.(4) The combination assay:when compared with those in endometriosis group,the ROC-AUC was 0.936,the specificity was 95% and the sensitivity was 81.0% in ovarian cancer.Conclusions Measurement of HE4 could be used in differential diagnosis of endometriosis cyst.And the combination of HE4 and CA125 assay could discriminate ovarian endometriosis cysts from ovarian malignant tumors effectively.

Objective To observe the effects of short hairpin RNA (shRNA) targeting Her2 on its gene expression when the shRNA was stably transfected into human ovarian cell lines, SKOV3 and SKOV3. ipl, which have different extent of malignancy and investigate the changes of the biological characters of the two cell lines after the stable transfection. Methods The plasmids expressing shRNA targeting Her2 gene were transfected into SKOV3 and SKOV3. ipl cells. The stably transfected cells were gained by antibiotic screening. The expression of Her2 before and after the transfection was detected by RT- PCR and western blot. The transwell experiment was used to observe the invasion ability of the cancer cells before and after the transfection, and the parent and the transfected cells were injected into nude mice to observe the tumor growth. Results After the stable transfection with Her2 shRNA, mRNA and protein levels of Her2 gene in SKOV3 and SKOV3. ipl cells were remarkably reduced. The expression of mRNA were (68.0±3. 1) %, (40. 8±2. 0) %, (99. 9±1.3) %, (42. 4±2. 5) %. The expression of protein were (72. 1±3.4) %, (36. 4±1.5) %, (98.2±1.7) %, (40. 7±2. 1) %. The invasion ability into basilar membrane of the transfected cells was greatly reduced compared with the parent cells. The invasion cell numbers were 7.6±1.1, 1.8±0. 8, 36. 2±9.7, 15.7±7. 2. The growth rate of the planted tumors was lower in transfected groups than that of the parent groups. Conclusions (1) The expression of Her2 gene in SKOV3 and SKOV3. ipl cells was remarkably reduced by RNA interference targeting Her2. (2) The biological characters of SKOV3 and SKOV3. ipl cells are changed when the expression of Her2 gene is reduced by RNA interference.

Objective:To compare the clinical and prognostic characteristics of ovarian endometrioid carcinoma (OEC) patients with synchronous endometrial lesions and patients with pure OEC.Methods:A retrospective review of the medical records of patients received initial treatment and a postoperative pathological diagnosis of OEC at Peking University People′s Hospital between August 1998 and December 2017 were performed. According to the inclusion criteria, a total of 56 patients with OEC were included in the study, including 13 patients concurrent with simultaneous endometrial lesions (Group A) and 43 patients with pure OEC (Group B).Results:Patients with synchronous endometrial lesions accounted for 23% (13/56). Mean age of Group A at diagnosis was (44.9±8.3) years old, 2/13 of patients were postmenopausal, and no one had a history of hypertension, the first symptom of 5/13 people was irregular vaginal bleeding. Mean age of Group B patients at diagnosis was (52.7±10.2) years old, 53% (23/43) of patients were postmenopausal, and 28% (12/43) patients had the history of hypertension, the first symptom of 4 (9%, 4/43) people was irregular vaginal bleeding. The differences of age, menopause status, history of hypertension and initial symptoms between the two groups were statistically significant (all P<0.05). There were no significant differences in fertility history, dysmenorrhea history, age of menarche, history of endometriosis, preoperative and postoperative CA 125 level, International Federation of Gynecology and Obstetrics (FIGO) stage, tumor grade, metastatic site and platinum-based chemotherapy drug resistance between the two groups (all P>0.05). The overall 5-year survival rate of OEC patients was 91.6%, and the overall 5-year progression-free survival rate was 76.6%. Among them, the 5-year survival rate of the OEC concurrent with simultaneous endometrial lesions group was 80.2%, and the pure OEC group was 93.4%; the 5-year progression-free survival rate of the OEC concurrent with simultaneous endometrial lesions group was 74.1%, and the 5-year progression-free survival rate of the pure OEC group was 77.3%. There were no significant differences between the two groups (all P>0.05). Multivariate analysis showed that the independent factors for the prognosis of OEC patients were FIGO stage ( P=0.006) and residual lesion size ( P=0.020). Conclusions:OEC patients have a high proportion of simultaneous endometrial lesions. OEC with simultaneous endometrial lesions are younger than patients with pure OEC. Synchronous endometrial lesions do not affect the prognosis of patients with OEC.

Objective To evaluate the value of human epididymis secretory protein 4(HE4)and CAl25 in the diagnosis of ovariall malignancy.Methods HF4 and CA125 in the serum specimens of malignant ovarian tumor group(30 cases),benign ovarian diseases(110 cases;45 benign ovarian tumor,57endometriotic diseases and 8 pelvic inflammation were included) and healthy women group( 137 cases)were assayed double blindly . The levels and the diagnosis efficiency of the HE4 and CA125 were analyzed.Results (1) The median levels of HE4 and CA125 were significantly higher in malignant ovarian tumor group (244 pmoi/L and 601 kU/L respectively) than those of the benign ovarian diseases group( 32 pmol/L and 22 kU/L respectively)and healthy women group (32 pmoi/L and 11 kU/L respectively) (P =0. 000-0. 029). The median levels of CA125 were also higher in endometriotic diseases and pelvic inflammation groups(53 and 41 kU/L respectively) than those of benign ovarian tumor group and healthy women group (12 and 11 kU/L respectively;P = 0. 000-0. 031 ). (2) The positive rate of HE4 was lower than that of CA12s in malignant ovarian tumor group ( P = 0. 036 ). HE4 was negative in benign diseases and healthy women groups. But the positive rates of CA125 were 56. 1% and 5/8 respectively in endometriotic diseases and pelvic inflammation groups and there were significant differences compared with HE4( P =0. 000). (3)The HE4 assay had advantage over the CA125 assay in receiver operating characteristic-area under the curve (ROC-AUC) and sensitivity with a specificity of 100% when ovarian malignancy was compared with controls having benign diseases and healthy women, benign tumor or benign diseases groups respectively. The CA125 assay had advantage over the HE4 assay in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having healthy women group. (4) Combined assay of HE4 and CA125was better than CA125 alone when ovarian malignancy was compared with controls having any group. (5)Combined assay was better than HE4 alone in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having benign diseases and healthy women or healthy women groups. And combined assay was lower in the ROC-AUC and the sensitivity with specificity of 100% than HE4 when ovarian cancers were compared with controls having benign tumors or benign diseases groups respectively. (6) The diagnosis efficiency of the HE4 assay at the level 86 pmol/L determined in ROC curve with controls having benign diseases and healthy women group and at the 95% reference level 50 pmol/L of healthy women or 150 pmol/L recommended by the kit respectively was compared. The sensitivity of 50 pmol/L was 73% higher than 150 pmol/L and 86 pmoi/L, while the specificity and positive predictive value were lower ( P = 0. 002, P = 0. 000 ). The specificity, accuracy and positive predictive value of HE4 assay at the set point of 150 pmol/L and 86 pmol/L were 100%, 96% and 96%. The set point of 86 pmol/L had advantage over 150 pmol/L at the sensitivity of diagnosis, 70% and 63% respectively. But the positive predictive value was 95% lower than 150 pmol/L, being 100%. There was no significant difference( P =0. 883, P = 0. 883 ). Conclusions The specificity of HF4 assay is higher than CA125 assay in the diagnosis of ovarian cancer and HE4 combined with CA125 assay can improve the diagnoses. The set point of 150 pmol/L is advantageous for the accurate diagnosis, while the set point of 86 pmol/L is advantageous for the screening of malignant ovarian cancer.

Objective:To investigate the association of time in range(TIR) with the severity of coronary artery disease and acute coronary syndrome in patients with type 2 diabetes mellitus.Methods:A total of 216 patients with type 2 diabetes mellitus and coronary heart disease were recruited and undergone anthropometric and biochemical measurements, continuous glucose monitoring, and calculation of SYNTAX score. TIR was defined as the percentage of time within the glucose range of 3.9-10.0 mmol/L during 24 h. Spearman correlation analysis and multivariate linear regression analysis were used to evaluate the correlation factors of SYNTAX score. Multivariate logistic regression analysis was used to analyze the association of TIR with the severity of coronary artery disease and acute coronary syndrome. Results:Compared with patients with mild coronary artery disease, TIR in patients with moderate to severe coronary artery disease was lower[(69.4±17.3)% vs (60.8±17.8)%, t=3.0, P=0.003], and HbA 1C of patients with moderate to severe coronary artery disease was higher [(9.6±1.7)% vs (8.8±1.6)%, t=3.3, P=0.001]. SYNTAX score was negatively correlated with TIR ( r=-0.251, P<0.01) and positively correlated with HbA 1C ( r=0.249, P<0.01). Moreover, compared with HbA 1C (standardized coefficients=0.181, P=0.007), TIR (standardized coefficients=-0.192, P=0.004) had a greater influence on SYNTAX score. Multivariate linear regression analysis showed that TIR, HbA 1C, duration of diabetes and smoking were independently correlated with SYNTAX score. Multivariate logistic regression analysis revealed that compared with TIR Q1, Q3 and Q4 were independent protective factors for moderate to severe coronary artery disease (respectively, OR=0.61 and 0.59, 95% CI 0.39-0.96 and 0.38-0.94, P=0.014 and 0.009) and acute coronary syndrome (respectively, OR=0.51 and 0.39, 95% CI 0.32-0.95 and 0.26-0.75, P=0.022 and 0.008). Conclusion:TIR was significantly and independently correlated with the severity of coronary artery disease and acute coronary syndrome in type 2 diabetes mellitus after controlling confounding factors. When TIR level was decreased, the severity of coronary artery disease was aggravated, and SYNTAX score and the risk of acute coronary syndrome was increased.