Objective To investigate the role of Ca~(2+) in the mechanism of ischemic preconditioning effects.Methods Fifty Sprague-Dawley male rats were divided into 5 groups randomly:IC group,ischemic control,IP group,ischemia followed by reperfusion;IP+Verapamil group,Bay k8644 group,Verapamil group.Results Siginificant heart functional recovery was observed in IP group as campared with that in IC group(P

Objective To determine the levels of blood pancreatic stone protein/ regenerating protein(PSP/reg)and to explore the value of PSP/ reg in assessing severity and predicting prognosis of sepsis with or without panc-reatic damage. Methods In this prospective study,the clinical data and blood samples to measure PSP/ reg levels in 1,3 and 7 days after admission in the Pediatric Intensive Care Unit,Hunan Children's Hospital,from the children with sepsis from September 2013 to September 2014 were collected. According to levels of pancreatic trypsin(amylase or li-pase),the children were divided into normal group,mildly elevated group(1 - 3 times),and severely elevated group (more than 3 times);based on the condition of sepsis,the children were divided into sepsis group and severe sepsis group. Chi - square test,nonparametric rank sum test,Spearman correlation and area under the receiver operating cha-racteristic curve(AUC),Kaplan - Meier survival curves were used for statistical analysis. Results (1)A total of 226 children with sepsis were included in this study,among them pancreatic trypsin was normal in 159 cases(70. 4% ), pancreatic trypsin mildly elevated in 44 cases( 19. 4% ),and pancreatic trypsin severely elevated in 23 cases (10. 2% );125 cases(55. 3% )were sepsis,and 101 cases(44. 7% ,of which 29 cases were sepsis shock)were severe sepsis.(2)The correlation between PSP/ reg and serum amylase was maximum,while PSP/ reg had some correlation with serum lipase,but there was no correlation with C - peptide,insulin,glucose.(3)When pancreatic trypsin was normal or mildly elevated,severe sepsis group had higher levels of PSP/ reg than that of sepsis group(Z = - 7. 627,- 2. 965,P =0. 000,0. 003);while pancreatic trypsin was severe elevated,there was no significant difference in PSP/ reg levels be-tween sepsis group and severe sepsis group(Z = - 0. 134,P = 0. 894).(4)The AUC of PSP/ reg and procalcitonin to assess severity of sepsis were greater than 0. 7 in the children whose pancreatic trypsin was normal or mildly elevated, and there was no significant difference between them(P ﹥ 0. 05);while the AUC of C - reactive protein and WBC were less than 0. 7.(5)PSP/ reg levels gradually increased in non - survival children in 3 and 7 days,while survival children gradually declined. The AUC of PSP/ reg to predict prognosis was 0. 775,and 60 μg/ L was viewed as the critical point, sensitivity 89% ,specificity 67% . Children with PSP/ reg levels ≤60 μg/ L had higher rate of survival than children with ﹥ 60 μg/ L(χ2 = 36. 874,P = 0. 000). Conclusions PSP/ reg is closely associated with pancreatic exocrine func-tion,but PSP/ reg still can assess severity and predict prognosis in children with sepsis and pancreatic damage.

Purpose To detect the HER2 gene amplification by fluorescence in-situ hybridization(FISH) and to explore its clinicopathologic relationship with the breast cancer.Methods A prospective study was conducted in 50 cases of breast invasive ductal carcinoma from Beijing Toren Hospital. Clinicopathologic data were summarized and FISH was performed in the paraffin-embedded sections for HER2 gene amplificayion using DNA probe, and immunohistochemical stain for ER, PR and HER2.Results The average age of the patients was 55.5 years. The pathologic grading showed that 11 cases were in grade Ⅰ, 30 cases in grade Ⅱ, and 9 cases in grade Ⅲ. The TNM staging showed that 13 cases were in stage Ⅰ,15 cases in ⅡA,13 cases in ⅡB,6 cases in ⅢA,2 cases in ⅢB,and 1 case in ⅢC.The median metastasis rate of lymph node was 6.91%.33 cases were positive for ER,and 32 cases positive for PR.For HER2 detection, 40 cases were positive by IHC and 33 positive by FISH. HER2 gene amplification by FISH was closely related with the expression of HER2 protein by immunohistochemistry, but not significantly related with pathologic grading, The TNM staging, median lymph node metastasis rate, ER and PR status (P>0.05). FISH test was positive in 3 cases of tumor embolus and 3 cases of multiple primary tumors. Conclusions FISH and immunohistochemistry for detecting HER2 have a good conformance, HER2 gene amplification may be related with tumor embolus and multiple primary tumors, but it can not be used as an indirect marker to predict the prognosis.

Objective To explore the potential mechanism of abnormal behavior resulted from maternal separation in neo-natal period in rat. Methods Neonatal rats were equally and randomly divided into maternal separation group and control group. The rats in maternal separation group were separated from the dam for 3h per day on postnatal days (PND) 2 to 21, nothing was done to the rats in the control group. The brain tissues were taken out after being killed on PND 7, 14, and 21. The expressions of Caveolin-l, BDNF and GFAP in hippocampal formation were detected by immunohistochemistry. Semiquantitative assessment of immunohistochemical images was performed by Image-Pro Plus software. Results Compared with control groups, the expres-sion of Caveolin-l on PND 7 had no signiifcant change, while BDNF and GFAP were signiifcantly increased in maternal separa-tion group (P<0.05). On PND 14 and 21, the expressions of Caveolin-l, BDNF and GFAP were signiifcantly decreased in maternal separation group (P<0.05). Conclusions Decreased expressions of Caveolin-l, BDNF and GFAP caused by maternal separation in neonatal period may be associated with abnormal behaviors in adulthood in rat.

ObjectiveTo analyze the expression of osteopontin (OPN) and interleukin-17 (IL-17)and study the roles in the pathogenesis of chronic severe hepatitis B.MethodsTwenty patients of acute on chronic liver failure were collected from the infection disease in-patients department of the First Affiliated Hospital of Medical College,Zhejiang University from 2009 to 2010,and 40 health controls were from medical examination center during the same period.Meanwhile,Balb/C mice were used for ConA injection to induce fulminant hepatitis and the plasma,serum and liver tissue of mice were collected.OPN and IL-17concentrations were measured using ELISA kits.PBMC were separated and cultured with anti-TNF-α or TNF-α.Supernatants were collected for analysis of OPN and IL-17.Differences between groups were evaluated by using a Student's t test and the relationship between IL-17 and OPN were evaluated by Pearson correlation.ResultsIn chronic severe hepatitis B group,levels of OPN and IL-17 were markedly higher than those of healthy control,respectively.(42.4 ± 12.9 vs 10.6 ±4.8 pg/ml; 1460.1 ±523.1 vs 472.8 ±67.5 ng/ml) ( t =2.387,3.570,P ＜ 0.05).The level of OPN in blood and liver reached peak at 6 hours at 12 hours after ConA injection,respectively.The level of IL-17 in blood and liver was significantly increased after ConA injection.IL-17 were positive correlated with OPN levels (R2 =0.582,P =0.026).TNF-α can increase the level of OPN secreted by lymphocytes.ConclusionsOPN and IL-17 levels in peripheral blood of hepatitis B patients are closely related to the hepatitis B degree.TNF-α can increase the level of OPN secreted by lymphocytes.

Objective To study the effects of modified Danggui Beimu Kushen Pills on the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissues on MFC gastric cancer bearing mice; To discuss relevant mechanism of action. Methods MFC gastric cancer bearing mice were employed to perform anti-tumor experiment in vivo in this study. A total of eligible 48 mice were randomly divided into model group, DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups, modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. The treatment was conducted once a day, and lasted for 14 continuous days. After the last administration of gavage orally treatment, all mice were anaesthetized and killed by cervical dislocation method to obtain tumor tissue completely for further HE staining measure and detection of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissue with the method of RT-qPCR and immunohistochemistry. Meanwhile, the tumor growth was observed and the general conditions of mice were recorded. Results The model group was rich in tumor cells; the sizes of cells were different; the volume was large; the nucleus was deeply stained and the heterotypic shape was obvious, and the small focal necrosis was seen. The number of tumor cells in each administration group was reduced; the arrangement was loose; the cell volume was significantly reduced, and the nuclear pyknosis was reduced. Cell necrosis significantly increased; the number of interstitial blood vessels decreased; collagen fibers increased, especially in modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. Compared with the model group, the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 mRNA and protein decreased in each administration group. TLR2, TLR4, TLR6, TRAF6 and MyD88 were lighter and weakly positive expressed in modified Danggui Beimu Kushen Pillshigh-dose and low-dose combined with DDP groups, the protein changes were more obvious Compared with DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups. Conclusion Modified Danggui Beimu Kushen Pills can down-regulate TLR2, TLR4, TLR6, TRAF6 and MyD88 expression of tumor tissue in MFC gastric cancer bearing mice at both mRNA and protein levels to play anti-tumor pharmacology action.

This study was aimed to look for the potential application of human umbilical plasma (HUP) in the culture of human umbilical vein endothelial cells (HUVECs). The effect of HUP concentration in cell culture medium on cell proliferation activity and cell cycle was studied. HUVECs were obtained by digesting the umbilical cord with 0.25% trypsin mixed with the equal volume of 0.1% collagenase II, then were identified by morphology and factor VIII immunohistochemistry under phase contrast microscopy. The cells cultured after 7 days showed the typical cobblestone morphology with factor VIII immunohistochemical staining positive. The study showed that the groups of 20% (HUP), 15% HUP+20% Fetal Bovine Serum (FBS) and 20% HUP+20%FBS enhanced cell proliferation activity significantly when compared with the control group (20% FBS without HUP). On the contrary, 30% HUP+20% FBS caused cell cycle arrest, which significantly hindered the proliferation of HUVECs. The study proved that although HUP might not be able to completely replace the role of vascular endothelial growth factor in cell culture, as a supplement ingredient, it was an ideal candidate to replace FBS in culture medium.
Cell Culture Techniques ; methods ; Cell Proliferation ; Cells, Cultured ; Culture Media ; chemistry ; Fetal Blood ; chemistry ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Plasma ; physiology

Thyroid dysfunction refers to the abnormality of thyroid hormone in the blood circulation .Hyper-thyroidism and hypothyroidism are the main clinical factors .Thyroid hormone can accelerate degradation of insulin , and can reduce the sensitivity of the organism (including the liver and peripheral tissues ) to insulin,moreover,thyroid hormone still can adjust the function of islet beta cells .Some studies have pointed out that the disorder of thyroid hormone has a certain influence on the incidence of insulin resistance .The purpose of this paper is to review the relationship between thyroid dysfunction and insulin resistance .

Objective:To analyze the reflux parameters of patients with gastroesophageal reflux disease (GERD) in upright position, supine position and at 2 h after meals, and to explore the cut-off value, sensitivity and specificity of the reflux parameters in different positions and at 2 h after meals in GERD diagnosis.Methods:From January 2016 to July 2020, 200 GERD patients (GERD group) and 61 non-GERD patients (control group) who visited Huazhong University of Science and Technology Union Shenzhen Hospital (Former Nanshan District People′s Hospital), were selected. All the patients of the two groups received gastroesophageal reflux disease questionnaire (GERDQ), upper gastrointestinal endoscopy, esophageal high resolution manometry and 24 h esophageal pH combined impedance monitoring. T test, non-parametric test and chi-square test were used to compare the related parameters in upright position, supine position and at 2 h after meals between two groups and within each group. Receiver oparative characteristic (ROC) curves of reflux parameters in upright position, supine position and 2 h after meals were drawn to determine the cut-off value, sensitivity and specificity in GERD diagnosis. Results:The proportion of patients with acid reflux in supine position of the control group was higher than that of the GERD group (41.0%, 25/61 vs. 8.50%, 17/200), and the difference was statistically significant ( χ2=36.53, P<0.01). In the control group, the acid reflux time in upright position, number of acid reflux, acid exposure time (AET), longest reflux time and number of weak acid reflux were more than those of in supine position in the same group (6.00 min(2.00 min, 13.50 min) vs. 0.00 min(0.00 min, 1.50 min), 16.00(8.00, 27.00) vs. 1.00(0.00, 3.00), 0.90%(0.33%, 1.88%) vs. 0.00%(0.00%, 0.30%), 2.00 min(1.00 min, 4.00 min) vs. 0.00 min(0.00 min, 1.00 min), 7.00(3.00, 11.00) vs. 1.00(0.00, 2.00), respectively) and the differences were statistically significant ( Z=5.43, 6.61, 5.06, 3.58 and 6.24, all P<0.01). In the GERD group, the acid reflux time, number of acid reflux, AET, longest reflux time and number of weak acid reflux in upright position were higher than those in supine position (51.00 min, (31.00 min, 86.75 min) vs. 8.00 min(1.00 min, 42.00 min), 60.00(48.00, 83.75) vs.6.00(2.00, 19.50), 7.30%(3.90%, 12.10%) vs. 1.50%(0.20%, 6.50%), 7.00 min(4.00, 12.00 min) vs. 4.00 min(1.00 min, 17.00 min), 1.00(0.00, 3.00) vs. 0.00(0.00, 2.00), 7.00(3.00, 12.00) vs. 0.00(0.00, 1.00), respectively) and the differences were statistically significant ( Z=7.92, 11.22, 6.90, 2.56, 5.11 and 11.76, all P<0.05). The acid reflux time, number of acid reflux, AET, longest reflux time and number of weak acid reflux at 2 h postprandial were 3.00 min(2.00 min, 9.00 min), 10.00(5.00, 18.00), 0.90%(0.40%, 1.98%), 1.00 min(0.00 min, 3.00 min), 4.00(1.50, 8.50)and 28.50 min(15.00 min, 54.75 min), 35.00(24.00, 52.00), 8.30%(4.32%, 15.83%), 6.00 min(3.00 min, 11.00 min), 4.00(2.00, 7.25), in the control and GERD groups, respectively, which were significantly higher than those in supine position in the same group ( Z=4.30, 6.33, 5.50, 3.40, 5.71 and 3.76, 9.21, 5.76, 1.97, 10.46, all P<0.05). Among 200 GERD patients, 125 patients had symptoms recorded during the 24 h esophageal pH combined impedance monitoring, the incidence of reflux symptoms in upright position was higher than that in supine position (89.6%, 112/125 vs. 65.6%, 82/125), and the difference was statistically significant ( χ2=20.71, P<0.01). The results of ROC curve analysis showed that the accuracy of acid reflux time in upright position in GERD prediction was the highest, with AUC value of 0.94 and cut-off value of 24.5 min, and the sensitivity and specificity in GERD diagnosis were 81.50% and 95.08%, respectively. The prediction accuracy of acid reflux times in upright position and AET in upright position for GERD was secondary, AUC value both were 0.93 and the cut-off value of the acid reflux number in upright position was 39.5, and the sensitivity and specificity in GERD diagnosis were 84.00% and 95.08%, respectively. The cut-off value of AET in upright position was 2.75%, the sensitivity and specificity in GERD diagnosis were 85.00% and 93.33%, respectively. The AUC value, cut-off value, sensitivity and specificity of AET at 2 h postprandial were 0.91, 4.60%, and 73.49% and 95.00%, respectively. Conclusions:Both GERD patients and non-GERD patients have more reflux in upright position, especially within 2 h after meals. The diagnostic values of acid reflux time in upright position, number of acid reflux, AET and AET 2 h after meals for GERD is high, and the AUC values are all >0.90, which can be used as a more comprehensive basis for the analysis and diagnosis of GERD.

Objective:To construct specific small interfering RNA(siRNA) expressing vectors of intracellular Ca2+ transport protein(CaT1)gene and detect its silencing effects.Methods:The hairpin sequences of siRNAs targeting CaT1 gene were designed,synthesized and cloned into pSlincer 3.1-H1 plasmids after annealing.The vectors were then enriched in E.coli.The recombinant pSlincer 3.1-H1 plasmids were identified by restriction endonuclease cutting and DNA sequencing and then transfected into Human Gastric Carcinoma Cell Line BGC-823.The expression of CaT1 mRNA was examined by RT-PCR.Results:The siRNA oligonucleotides of CaT1 were correctly cloned into the pSlincer 3.1-H1 plasmids and confirmed by restriction endonuclease cutting and DNA sequencing.RT-PCR analysis revealed that the expression of CaT1 mRNA in BGC-823 cells transfected with the pSlincer 3.1-H1 constructs of siRNA was significantly decreased compared with that of the negative control and untransfected group.Conclusion:siRNA expression plasmids for silencing Ca2+ transport protein gene are successfully constructed,and they effectively inhibit the CaT1 gene expression.