Objective:To find out markers of endometriosis. Methods: The two dimensional gel images of proteins extracted from eutopic endometrium from endometriosis patients and controls were analyzed by software Phoretix 2D,and the proteins expressed differently were identified primarily by query of data base.The proteins extracted from ectopic endometrium of ovarian endometriosis were transferred from two dimensional gel onto nitrocellulose membranes,followed by incubation with sera from women with and without endometriosis. Analyzed by MALDI-TOF-MS,the proteins hybridized differently were identified through their Peptide Mass Footprints. Results: Having compared the reproducible two dimensional gel images of proteins from eutopic endometrium of women with and without endometriosis,we obtained 11 proteins expressed differently.Through Western Blot technique,we found three proteins hybridized differently which were identified as vimentin,?-actin and ATP synthase ? subunit respectively. Conclusion: The protein expression spectra of eutopic endometrium from patients with endometriosis are significantly different from those of the controls,and the anti-endometrial autoantibodies against vimentin, ?-actin and ATP synthase ? subunit may be induced.

Objective To explore the infections of human cytomegalovirus (HCMV) in chronic hepatitis C patients and the hepatic impairment in chronic hepatitis C patients co-infected with HCMV.Methods HCMV-DNA was determined by fluorescence quantitative-PCR (FQ-PCR) in 95 patients with chronic hepatitis C (observation group) and 95 healthy controls(control group) and HCMV active infections were analyzed.HCV-RNA was determined by FQ-PCR in observation group,and the difference of HCMV-DNA positive rate between high HCV-RNA(＞ 104 copies/ml) and low HCV-RNA(≤ 104 copies/ml) was analyzed.Alanine aminotransferase (ALT),aspartate aminotransferase (AST) were determined by rate method in two groups and the hepatic impairment was analyzed.Results Twenty-five cases with positive HCMV-DNA in observation group,the positive rate was 26.3％(25/95).Five cases with positive HCMV-DNA in control group,the positive rate was 5.3％(5/95).There was significant difference between two groups for HCMV-DNA (x2 =14.29,P ＜0.01).Twenty-one cases with positive HCMV-DNA in 43 cases of high HCV-RNA patients,the positive rate was 48.8％(21/43).Four cases with positive HCMV-DNA in 52 cases of low HCV-RNA patients,the positive rate was 7.7％(4/52).There was significant difference between the two (x2 =19.90,P ＜ 0.01).ALT,AST in observation group was higher than that in control group (P ＜ 0.01).ALT,AST in chronic hepatitis C patients positive for HCMV-DNA was higher than that in chronic hepatitis C patients negative for HCMV-DNA significantly (P ＜ 0.01).Conclusions HCMV in chronic hepatitis C patients becomes active again and co-infects easily.When chronic hepatitis C patients co-infect HCMV actively,hepatic is further injured.

Objective To investigate the mechanisms modulating the functions of dendritic cells (DCs) and suppressing the activation and proliferation of T cells by transforming growth factor-β (TGF-β).Methods Mouse splenic DCs were purified with CD11c+ immunomagnetic beads and the purity of isolated DCs were detected by flow cytometry.Gene chip was used to detect gene expression in DCs after stimulation with TGF-β, and then real-time PCR was performed to analyze the differentially expressed genes in microarray at mRNA level.The activation and proliferation of CD4+ T cells which were co-cultured with DCs after stimulation with TGF-β were detected by flow cytometry.Results The purity of DCs reached over 95% after isolation.TGF-β down-regulated the expression of cell surface markers CD53, CD69, CD33, CD74 and CD93 on DCs;decreased the expression of chemokines Ccl3, Ccl5, Ccl9, Ccl6, Ccl17, Cxcl10, Ccl22, Ccl4, Ccr7, Ccl2, Cxcl9 and Ccl7;inhibited the expression of inflammatory cytokines IL-2ra, IL-12rb2, IL-15ra, IL-1b and IL-15.Moreover, the DCs-mediated activation and proliferation of CD4+ T cells were suppressed by TGF-β.Conclusion TGF-β inhibits the DCs-mediated activation and proliferation of CD4+ T cells by suppressing the expression of surface markers on DCs and down-regulating the expression of chemokines and inflammatory cytokines.

Objective To explore the regulation function of Icariine on the expression of bone morphogenetic protein signaling pathway Smad1,Smad5,Smad4 and to explore the mechanism of promoting MC3T3-E1 cell proliferation and differentiation.Methods MC3T3-E1 cells were treated by 0,10-9,10-8 and 10-7 mol/L Icariine respectively.After stimulated by Icariine 1 d,2 d and 3 d,MTT method and population diploid time were used to observe the cell proliferation,and the cell alkaline phosphatase (ALP) level was assayed.At 21 days later,the alizarin red staining was proceeded.At 1,2 and 3 days later,the RT-PCR was used to detect the mRNA expression level about Smad1,Smad5 and Smad4,and the Western blot was to detect the Smad1,5 and Smad4 protein.At 2 days later,the RT-PCR was used to detect the mRNA expression level about Runx2,BMP-2 and osteoprotegerin (OPG),and the Western blot was used to analyze osteocalcin (OCN) protein level.Results After simulated by Icariine,the proliferation (MTT test),the ALP activity and mineralization of osteoblasts were increased,the cell population diploid was reduced (P＜0.05).At 1,2 and 3days later,the results of RT-PCR showed that Icariine continued increasing the mRNA level of Smad1,5 and Smad4 in 10-8 mol/L.At 2 days later,Smad1,Smad5 and Smad4 mRNA expression were obviously reduced in 0 mol/L group,and At 3 days later,Smad1,Smad5 and Smad4 mRNA expression were obviously reduced in 10-9 and 10-7 mol/L groups.At 2 days later,BMP-2,Runx2 and OPG mRNA were obviously increased in 10-8 mol/L group.The results of Western blot showed that OCN,Smad1,Smad5 and Smad4 protein were obviously up-regulated in 10-8 mol/L group.Conclusion Icariine occurs the expressions of BMP-2,Runx2,Smad1,Smad5 and Smad4 by stimulating the bone morphology of MC3T3-E1 cells directly,and promotes the osteogenic differentiation in the manner of expression level synchronous rising.

Objective To investigate clinical features of lower motor neuron lesion (LMNL) caused by the single level lower thoracic disc protrusion (LTDP),and to observe clinical outcomes of surgical treatment.Methods Between January 1997 and December 2009,17 patients with LMNL caused by single level LTDP underwent en bloc resection of the superior articular process,Cave-in 360° circumferential decompression and internal fixation in our hospital.MRI and CT scans were taken to confirm lesion levels:T10-11 in 4 patients of whom 3 had patellar clonus and ankle clonus,T11-12 in 5 patients of whom 4 had ankle clonus,and T12L1 in 8 patients who only had positive Babinski sign.The neurologic status was assessed using the Japanese Orthopaedic Association (JOA) scoring system.The muscle strength of the tibialis anterior was assessed using the Manual Muscle Test (MMT).Sagittal Cobb angle and cross-sectional area of the dural sac at the level of maximal compression in MRI were also observed.Results All patients were followed up for 22 to 76 months (average,48.6 months).The mean JOA score increased from preoperative 5.88±1.11 to 9.53±0.94 at final follow-up (t=16.143,P＜0.05).The muscle strength of the tibialis anterior recovered to more than grade 4 in all patients.Postoperative Cobb angle was unchanged compared with that before operation.MRI indicated that the cross-sectional area of the dural sac at the level of maximum compression increased from preoperative 35.8±7.3 mm2 to postoperative 132.9±6.5 mm2 (t=70.78,P＜0.05).Conclusion LMNL can be caused by LTDP.The eu bloc resection of the superior articular process,Cave-in 360° circumferential decompression and internal fixation can provide a satisfactory decompression effect and marked recovery of neurological function.

Objective To explore the roles of serum TLR-4, TNF-αand IL-6 in neonates with preterm birth. Methods A total of 120 neonates from neonatology department in the Xingtai People's Hospital were selected and divided into full-term group (n=40), premature rupture of fetal membranes (n=40) and idiopathic preterm group (n=40) based on the gestational age. The peripheral venous blood was collected within 30 minutes when the infants were born, and the supernatant was reserved after centrifuged. The levels of serum TLR-4, TNF-αand IL-6 were detected by enzyme-linked immunosorbent assay. Results The levels of TLR-4, TNF-αand IL-6 in idiopathic preterm and premature rupture of fetal membranes were signiifcantly higher than that in full-term group and showed positive correlation. Conclusion Cytokines TLR-4, TNF-αand IL-6 maybe closely related to the preterm birth.

Objective To analyze the incidence and survival trend of malignant tumors in urban residents of Shenyang from 2011 to 2018. Methods The Shenyang tumor registration report system was used to collect the onset data and survival data of patients with malignant tumor from 2011 to 2018.The crude incidence, age-standardized rate, cumulative rate (0-74 years old), truncated rate (35-64 years old), survival rate, and incidence and survival rank were calculated.The observed survival rate was calculated by the life table method.The expected survival rate and relative survival rate were calculated by EdererⅡmethod.Using Joinpoint 3.5.3 software, the annual percentage change in incidence rate and survival rate (APC%) were calculated.SPSS23.0 software was used for the chi square tests of males and females. Results The crude incidence of malignant tumors in Shenyang, age-standardized incidence rates by Chinese standard population and world standard population were 364.70/10 million, 190.00/10 million and 185.63/10 million, respectively.The cumulative rate (0-74 years old) was 21.17%, and the truncated rate (35-64 years old) was 311.66/10 million in the years 2011-2018.The top five incidence rates of males are lung, colorectal, liver, stomach, and bladder cancer, whereas those of females were breast, lung, colorectal, uterine, and thyroid cancer.The incidence rate of malignant tumors increased in 8 years (P=0.00, P=0.67), and the incidence rate was higher in males than that in females (χ²=201.63, P < 0.05).The 5-year survival rate of malignant tumors was 40.49%, and the relative survival rate was 47.84% from 2011 to 2015.The five survival rates of males were in the order of thyroid, kidney, bladder, colon-rectum, and prostate cancer.The five survival rates of females were in the order of thyroid, breast, uterus, cervix, and colon-rectum cancer.The 5-year survival rate showed an upward trend (APC%=7.41, P=0.04).The survival rate of females was higher than that of males (χ²=187.62, P < 0.05). Conclusion The incidence rate and survival rate of malignant tumor in Shenyang urban residents increase yearly from 2011 to 2018.The incidence rate of males is higher than that of females, and the survival rate of males is lower than that of females.The incidence rate and survival rate of tumors differ much in sequence.

Objective: To analyze the results obtained from the proficiency testing program for determination of bovine blood lead and urinary cadmium, so as to evaluate the proficiency of the laboratories, and improve the detection and quality control capability of occapational health laboratory. Methods: Prepared 3 levels of lead samples with bovine blood as base material, taking Technical Norm of Primary Reference Material as Operating standard, 60, 80, 180 μg/L were used as the standard concentration. All samples were made for 200 bottles. Normal urine were used to prepared 3 levels of cadmium samples with 5.0, 7.0, 10.0 μg/L as standard concentration, then 3 level of samples were made for 200 bottles respectively. Blood lead and urinary cadmium samples were sent to each laboratory in the form of sample pairs. One-way ANOVA was used to investigate the homogeneity. The results of each laboratory were statistically analyzed by robust statistical method, and the detection ability of the laboratory was evaluated by the Z ratio score method. By consulting the test reports and original records of each participating laboratory, the reasons for the unsatisfactory results were analyzed. Results: The statistical F of the uniformity of blood lead 1.42, 1.37 and 2.85, F<3.13, P were 0.294, 0.314 and 0.059, respectively in 31 labs. There is no statistical significance in the difference of blood lead (P>0.05). And urine cadmium was less than the critical value (P>0.05). In all, 31 laboratories participated in the test of blood lead, and 28 loboratories paticipated in urine cadmium verification. The statistical F of the uniformity of urinary cadmium 1.90, 1.35 and 1.19, F<3.13. The homogeneity of the samples were in accordance with the requirements of proficiency testing program. The satisfactory rate of the results for blood lead was 84% (26/31) , and the satisfactory rate of urinary cadmium was 93% (26/28) . In the test of blood lead, 21 of the 31 laboratories were controlled by the standard material method. Totally, 20 of them get an satisfactory level, the rate was 95%. Conclusion Most of the laboratory test results were satisfactory, which showed that the detection capability for blood lead and urine cadmium was good. The main causes of unsatisfactory results were detection methods, tools of quality control ortraceability standards.

Objective: To develop a method for determination of metabolites of diphenylmethane diisocyanate (MDI) in urine, i.e. methylenedianiline (MDA) by high performance liquid chromatography-tandem mass (LC-MS-MS) . Methods: Urine samples were prepared by hydrolyzation with sulfuric acid and extraction by acetonitrile, and then separated on a Shim-pack XR-ODS column, analyzed with high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) . The external solvent standard calibration were tested. Results: The linearity ranges were 0.05~20.00 μg/L, The related coefficients were 0.999 5. The limit of detection was 0.02 μg/L. The rats of recovery were 91.0%~103.4%. The relative standard deviations were between 2.7%~7.3%. Conclusion The method was sensitive, accurate and suitable for the MDA determination in urine of MDI exposed population.

Objective To summarize the computed tomography (CT) and magnetic resonance imaging (MRI) features of IgG4-related pancreatitis.Methods The retrospective and descriptive study was conducted.The clinical data of 23 patients with IgG4-associated pancreatitis who were admitted to Sir Run Run Shaw Hospital of Zhejiang University School of Medicine from November 2012 to May 2018 were collected.There were 21 males and 2 females,aged from 45 to 83 years,with an average age of 63 years.Patients underwent upper abdominal enhanced CT and enhanced MRI examinations.Observation indicators:(1) imaging examination of the patients;(2) imaging characteristics on CT and MRI examinations;(3) follow-up.Follow-up using outpatient examination including laboratory and imaging examination was performed to detect clinical symptoms and signs once a month within 3 months postoperatively,once every 3 months within 3 months to 1 year postoperatively,once every 6 months with 1-2 years postoperatively,and once a year after 2 years postoperatively up to August 2018.Measurement data with skewed distribution were represented as M (range).Count data were described as absolute numbers.Results (1) Imaging examination of the patients:19 of 23 patients underwent upper abdominal enhanced CT combined with upper abdominal enhanced MRI scanning,3 underwent upper abdominal enhanced CT combined with upper abdominal MRI plain scanning,and 1 underwent only upper abdominal enhanced MRI scanning.(2) Imaging characteristics on CT and MRI examinations:pancreatic manifestations included 7 aspects.① Location of lesion and morphological classification:of 23 patients,17 were diffuse type,showing sausage-like appearance;4 were focal type,including 2 with mass in the pancreatic head and 2 with mass in the pancreatic body and tail;2 were multiple type,including 1 with mass in the pancreatic head,body and tail,and 1 with mass in the pancreatic body and tail.② Density of lesions on CT examination:of 23 patients,22 received CT plain scanning,including 10 with equal density and 12 with slightly low density;1 didn't receive CT plain scanning.③ Signal of lesions on MRI examination:on T1 weighted imaging,16 of 23 patients showed homogeneous slightly low signal,4 showed isointensity signal,2 showed mixed slightly low signal,and 1 showed slightly high signal.On T2 weighted imaging,21 of 23 patients showed homogeneous slightly high signal,1 showed isointensity signal,and 1 showed mixed slightly high signal.Of 23 patients,19 underwent diffuse weighted imaging (DWI) and 4 didn't undergo DWI.There were 17 patients with slightly high signal and 2 with high signal on DWI.On apparent diffusion coefficient imaging,10 patients showed slightly low signal,and 9 showed low signal intensity.④ Calcification:of 23 patients,2 had multiple calcifications including 1 of diffuse type with calcification located at pancreatic head,and 1 of focal type with calcification located at pancreatic body and tail;20 had no calcification;1 without CT plain scanning cannot be judged calcification.⑤ Enhancement pattern:23 patients showed progressive delayed enhancement of pancreatic lesions on enhancement scanning,homogeneous or heterogeneous enhancement in the arterial phase,and further enhancement in the portal venous phase and delayed phase (enhancement degree of focal type and multiple type was basically consistent with that of normal pancreas).⑥ Halo sign surrounding lesions:of 23 patients,7 had no halo sign,including 2 of diffuse type,3 of focal type,and 2 of multiple type;16 had halo sign (15 of diffuse type and 1 of focal type),including 11 with halo sign surrounding pancreatic body and tail,3 with halo sign surrounding whole pancreas,and 1 with halo sign posterior pancreatic body and tail,and 1 with halo sign anterior pancreatic head.⑦ The main pancreatic duct:14 of 23 patients had irregular stenosis of the main pancreatic duct,7 had mild dilatation of the main pancreatic duct,and 2 had no dilatation of the main pancreatic duct.Extra-pancreatic manifestations included 3 aspects.① Bile duct dilatation:10 of 23 patients had simple intra-hepatic and extra-hepatic bile duct dilatation,8 had no intrahepatic or extra-hepatic bile duct dilatation,and 5 had intra-pancreatic common duct stenosis with upstream bile duct dilatation.② Extra-pancreatic organ involvement:18 of 23 patients had extra-pancreatic organ involvement (5 with gall bladder involvement,4 with intra-hepatic and extra-hepatic bile duct and gall bladder involvement,3 with intra-hepatic and extra-hepatic bile duct involvement,1 with lung involvement,1 with spleen involvement,1 with common duct involvement,1 with kidney involvement,1 with lung and intra-hepatic and extra-hepatic bile duct involvement,1 with fibrosis of mesenteric root),and 5 had no extra-pancreatic organ involvement.③ Retroperitoneal lymph nodes:2 of 23 patients had enlarged retroperitoneal lymph nodes,21 had no enlarged retroperitoneal lymph nodes.(3) Follow-up:of 23 patients,19 were followed up for 3-74 months,with a median time of 22 months.Sixteen of 19 patients had various degrees of improvement in clinical symptoms,laboratory and imaging manifestations after standardized hormonotherapy,and 3 had no response to hormonotherapy.Conclusions CT and MRI examinations have important diagnostic value for IgG4-related pancreatitis.The pancreas mainly show diffuse swelling and halo sign surrounding lesions is a typical imaging characteristic.