Objective:To observe the effect of clopidogrel bisulfate combined with isosorbide mononitrate on levels of extracellular matrix metalloproteinase inducer(EMMPRIN),free fatty acid(FFA)and salusin-β in patients with coronary heart disease(CHD).Methods:This randomized controlled study enrolled 120 CHD patients admitted Hai'an People's Hospital between September 2019 and March 2021.The patients were divided into control group(n=60,isosorbide mononitrate therapy)and combined treatment group(n=60,clopidogrel bisulfate combined with isosor-bide mononitrate).Therapeutic effect,cardiac function,hemorheology,levels of EMMPRIN,FFA and salusin-β,and incidence of adverse reactions were compared between two groups.Results:Total effective rate in combined treatment group was significantly higher than that of control group(90％vs.70％,P=0.006).After 3-month treatment,compared with patients in control group,those in combined treatment group had significant higher cardi-ac output(CO)[(6.37±0.42)L/min vs.(8.97±0.52)L/min],left ventricular ejection fraction(LVEF)[(56.85±6.28)％vs.(61.16±7.14)％]and stroke volume(SV)[(64.55±4.19)ml vs.(69.74±5.32)ml],and significant lower plasma viscosity[(1.58±0.36)mPa·s vs.(1.09±0.25)mPa·s],whole blood viscosity[(6.91±0.79)mPa·s vs.(5.27±0.84)mPa·s],hematocrit[(48.17±5.73)％vs.(42.28±5.88)％],levels of EMMPRIN[(1.31±0.19)vs.(1.08±0.18)],FFA[(0.73±0.16)mmol/L vs.(0.54±0.15)mmol/L]and salusin-β[(3.59±0.49)nmol/L vs.(2.87±0.42)nmol/L](P＜0.001 all).We detected no significant differ-ence in incidence of adverse reactions between two groups(P=0.591).Conclusion:Clopidogrel bisulfate combined with isosorbide mononitrate can effectively improve cardiac function and hemorheology,and reduce EMMPRIN,FFA and salusin-β levels in CHD patients,and the safety is good.

Objective:To observe the effect of clopidogrel bisulfate combined with isosorbide mononitrate on levels of extracellular matrix metalloproteinase inducer(EMMPRIN),free fatty acid(FFA)and salusin-β in patients with coronary heart disease(CHD).Methods:This randomized controlled study enrolled 120 CHD patients admitted Hai'an People's Hospital between September 2019 and March 2021.The patients were divided into control group(n=60,isosorbide mononitrate therapy)and combined treatment group(n=60,clopidogrel bisulfate combined with isosor-bide mononitrate).Therapeutic effect,cardiac function,hemorheology,levels of EMMPRIN,FFA and salusin-β,and incidence of adverse reactions were compared between two groups.Results:Total effective rate in combined treatment group was significantly higher than that of control group(90％vs.70％,P=0.006).After 3-month treatment,compared with patients in control group,those in combined treatment group had significant higher cardi-ac output(CO)[(6.37±0.42)L/min vs.(8.97±0.52)L/min],left ventricular ejection fraction(LVEF)[(56.85±6.28)％vs.(61.16±7.14)％]and stroke volume(SV)[(64.55±4.19)ml vs.(69.74±5.32)ml],and significant lower plasma viscosity[(1.58±0.36)mPa·s vs.(1.09±0.25)mPa·s],whole blood viscosity[(6.91±0.79)mPa·s vs.(5.27±0.84)mPa·s],hematocrit[(48.17±5.73)％vs.(42.28±5.88)％],levels of EMMPRIN[(1.31±0.19)vs.(1.08±0.18)],FFA[(0.73±0.16)mmol/L vs.(0.54±0.15)mmol/L]and salusin-β[(3.59±0.49)nmol/L vs.(2.87±0.42)nmol/L](P＜0.001 all).We detected no significant differ-ence in incidence of adverse reactions between two groups(P=0.591).Conclusion:Clopidogrel bisulfate combined with isosorbide mononitrate can effectively improve cardiac function and hemorheology,and reduce EMMPRIN,FFA and salusin-β levels in CHD patients,and the safety is good.

Objective:To assess the severity of hypoxic-ischemic brain damage (HIBD) in neonatal rats and predict the occurrence of subsequent neurobehavioral abnormalities after brain injury by scoring and magnetic resonance imaging (MRI).Methods:7-day-old of 60 Sprague-Dawley (SD) rats were randomly divided into control group (14 rats), sham operation group (14 rats) and HIBD model group (32 rats). HIBD model was established by right common carotid artery dissection with Rice-Vannucci method and hypoxia. Within 24 h after modeling, the rats in the model group were evaluated by general condition score and Longa score, and the surviving rats with moderate and severe HIBD were selected for the experiment. 24 h after modeling, 5 rats of the model group were randomly selected for 2,3,5-triphenyltetrazole chloride staining to verify cerebral infarction. 1 week after modeling, 6 rats from each group were randomly selected for hematoxylin-eosin staining to observe HIBD brain injury. 4 weeks after modeling, 4 rats were randomly selected from the control group and the sham operation group, and 8 rats from the remaining model group were used to evaluate the volume of brain damage by MRI. 5-6 weeks after modeling, the remaining 8 rats from each group were subjected to the Cylinder test, and at 13 weeks, they underwent the Morris water maze test to evaluate their neurobehavior.Results:In HIBD model group, 19 rats with moderate to severe HIBD were selected from 32 rats. 24 h after modeling, cerebral infarction was verified in all rats, indicating moderate to severe HIBD. Brain tissue pathology observed 1 week after modeling revealed predominantly gray matter brain damage. MRI showed that 7 out of 8 rats had moderate to severe HIBD. Compared to the control and sham operation groups, the model group exhibited a significant decrease in the usage rate of the left forelimb in the Cylinder test at 5-6 weeks after modeling ( P<0.05), and the latency period in Morris water maze test was significantly prolonged at 13 weeks after modeling ( P<0.05), and the times of crossing platform quadrant were significantly reduced ( P<0.05). There was no significant difference in the right brain injury volume between 24 h and 4 weeks model group ( P>0.05). The brain injury volume in model group was negatively correlated with the usage rate of left forelimb in cylinder test at 5-6 weeks and the times of crossing platform quadrant in Morris water maze test at 13 weeks ( P<0.05), and positively correlated with latency period in Morris water maze test at 13 weeks ( P<0.05). Conclusions:Within 24 h of HIBD modeling, the severity of brain injury can be preliminarily predicted by general condition score and Longa score. 4 weeks after modeling, in the chronic phase of brain injury, MRI was proved to be an excellent predictor for mid-term and long-term neurobehavioral abnormalities in HIBD rats.

Myocardial infarction (MI) is one of the leading causes of death in the world. With the improvement of clinical therapy, the mortality of acute MI has been significantly reduced. However, as for the long-term impact of MI on cardiac remodeling and cardiac function, there is no effective prevention and treatment measures. Erythropoietin (EPO), a glycoprotein cytokine essential to hematopoiesis, has anti-apoptotic and pro-angiogenetic effects. Studies have shown that EPO plays a protective role in cardiomyocytes in cardiovascular diseases, such as cardiac ischemia injury and heart failure. EPO has been demonstrated to protect ischemic myocardium and improve MI repair by promoting the activation of cardiac progenitor cells (CPCs). This study aimed to investigate whether EPO can promote MI repair by enhancing the activity of stem cell antigen 1 positive stem cells (Sca-1⁺ SCs). Darbepoetin alpha (a long-acting EPO analog, EPO_anlg) was injected into the border zone of MI in adult mice. Infarct size, cardiac remodeling and performance, cardiomyocyte apoptosis and microvessel density were measured. Lin^- Sca-1⁺ SCs were isolated from neonatal and adult mouse hearts by magnetic sorting technology, and were used to identify the colony forming ability and the effect of EPO, respectively. The results showed that, compared to MI alone, EPO_anlg reduced the infarct percentage, cardiomyocyte apoptosis ratio and left ventricular (LV) chamber dilatation, improved cardiac performance, and increased the numbers of coronary microvessels in vivo. In vitro, EPO increased the proliferation, migration and clone formation of Lin^- Sca-1⁺ SCs likely via the EPO receptor and downstream STAT-5/p38 MAPK signaling pathways. These results suggest that EPO participates in the repair process of MI by activating Sca-1⁺ SCs.
Animals ; Mice ; Ventricular Remodeling ; Erythropoietin ; Myocardial Infarction ; Heart ; Stem Cells

Aim To investigate the protective effect of mGluR5 activated by VU0360172 on germinal matrix hemorrhage in neonatal rats.Methods Seven day- old SD rats were randomly divided into Sham, GMH, and low-, medium-, and high-dose groups.The model was established by intracerebral injection of collagenase W-S.Then three doses of VU0360172 were injected intraperitoneal^ 3 h after surgery.Sham and GMH group were given the same amount of solvent.Neurobe- havioral tests were performed 24 h after surgery.Then the brain tissues were collected for evaluation of brain water content, brain hemoglobin content and HE stai¬ning.The expressions of Bcl-2 and cleaved-caspase-3 were determined by Western blot.Results Compared with Sham, GMH group had pooler behaviors in neuro- functional tests with increased brain water content and brain hemoglobin content (P < 0.01 ).And brain tis¬sues were destroyed significantly.WB results showed the expression level of Bcl-2 decreased ( P < 0.05 ) , while cleaved-caspase-3 being up-regulated ( P < 0.01).However, the administration of VU0360172 improved neurological function and ameliorated brain edema and hemorrhage ( P <0.01 ).Brain pathologi¬cal damage was reduced.Moreover, the stimulation of mGluR5 up-regulated Bcl-2 protein expression ( P < 0.05 ) and decreased the level of cleaved-caspase-3 ( P <0.01 ).Conclusion Activation of mGluR5 by VIJ0360172 protects against germinal matrix hemor¬rhage in neonatal rats.

Ischemic dysfunction is an important global health problem. Vascular endothelial cells(VECs) play a key role in angiogenesis, and insufficient vascular remodeling may lead to chronic nonhealing wounds. Therefore, effective VEC generation strategies of exploration help improve angiogenesisin damaged tissues. Embryonic stem cells (ESCs) are widely used in the study of tissueendothelialization, and endothelial progenitor cells (EPCs) are indispensable parts of the development ofVECs. The aims of this study were to find a rapid, easily screened and reproducible method for thederivation of EPCs from mouse embryonic stem cells (mESCs), and obtain VECs with high survival ratesand strong functions from the directed differentiation of the EPCs. The results showed that mESCs weredifferentiated into " stepping stone" -like progenitor cells with active proliferative ability by 10 ng / mLVEGF and 5 ng / mL bFGF. At the same time, the method of differential adherence was helpful for theselection of EPCs, and EPCs induced high expression of CD133 and CD34 (The relative expressionlevels were 0. 88 ± 0. 04 and 2. 12 ± 0. 02, respectively) for 3 days. Then EPCs were digested withacctuse enzymes, and induced to differentiate into vascular endothelial-like cells by 50 ng / mL VEGF and25 ng / mL bFGF for 7 days. The endothelial cells not only expressed endothelial marker genes (CD31, CD144, LAMA5, Tek, KDR and vWF),and marker proteins CD31, CD144 and LAMA5 (The relativeexpression levels were 1. 07 ± 0. 03, 0. 60 ± 0. 02 and 0. 70 ± 0. 02, respectively), but also had thegood ability of migration, tubulogenesis and formation of W-P bodies. Moreover, PBS, EPC and VECwere used to treat wounds of the same size. Both EPC and VEC could accelerate the degree of tissuehealing (The relative healing rates were 78. 93 ± 75. 35%, 95. 57 ± 83. 73% and 100. 00 ± 0. 00%, respectively), and VEC significantly enhanced the ability of wound angiogenesis and inflammatoryresponses. In consequence, this study preliminarily confirmed that mESC-derived EPCs coulddifferentiate into VECs after directional induction for 7 days, which had good function of tissue repair. The physiological pathway on stem cells by stimulating angiogenesis is expected to become a new target fortissue remodeling.

Objective To research the monoclonal antibody KMP1 inhibited bladder cancer EJ cell lines growth and metastasis in vivo by bioluminescence imaging. Methods Immunohistochemistry was used to determine the KMP1 binding to EJ and EJ-GFP cell lines. The xenograft tumor cell growth and distribution were measured by vernier calipers and dynamic in vivo fluorescence imaging. Immunohistochemistry and H&E counterstaining researched the feature of the xenograft tumor. Results Cell growth curves of EJ and EJ-GFP cells were similar. EJ-GFP had a green fluorescence. In EJ-GFP nude mouse tumor model, the addition of KMP1 significantly inhibited tumor growth and extended the average life span of nude mice. Both EJ and EJ-GFP cells can bind to KMP1,and the weight of transplanted tumors in the KMP1 treatment group was significantly lower than that of the mIgG control group (P＜0.001).Conclusion KMP1 has a promising antitumor effect in vivo. It might be valuable for development as a promising targeted agent for bladder cancer.

Objective By analysis of the key performance indexes of the clinical laboratory automation system, to clarify the advantage and optimize the comprehensive performance of the laboratory automation system.Methods Key performance indexes were Collected from January 2017 to April 2017 in biochemistry and immunoassay group of Clinical Laboratory of Shanghai Tong Ren Hospital.(1)The data were collected and compared by the before-and-after method,the starting time of the automation system and initial sample test were analyzed.(2)Key performance indexes were analyzed for the time of specimen registration,inspection,and reporting.(3)The specimen turnaround time(TAT)was analyzed based on two months operation of the laboratory automation system.In view of disadvantage of infectious assays, setting up priority sample absorption, then TAT performance was re-evaluated.(4)By the assessment of total serum dosage required in the automation system, the number of blood vacuum tubes were reduced reasonably.The pros and cons of laboratory automation system were analyzed and the potential improvement were proposed.Results (1)According to the sample peak shift forward,the system start time could move forward 30 minutes earlier.(2)With the adopting of railway logistics,the specimens were sent to the lab and the registration time was at 7:25 am,and the time required for specimen delivery was greatly reduced which made specimen test,report and audit time all moved forward accordingly.(3)Data has shown that specimen TAT declined dramatically based on the performance of the first two month operation of the automation system,biochemical items were shortened 2 h,and the immunoassay shortened 4 h,respectively.Moreover the trend keeps better gradually.With setting up priority absorption infectious tests,the TAT was improved greatly,TAT reduced the average by 40 min.(4)500 μl(including the sample in dead space of vacuum tube)were needed for all the 65 biochemical items included in the system, and 1 495 μl serum were used for the 28 immunoassay.As a result, a total of 2 000 μl serum will be enough for sample analysis by the system, which provided the feasibility to reduce 3 vacuum tubes averagely.Considering the current automation system does not include all the analysis items in our lab directory, a few tests remain to be performed on offline instruments respectively.The methodology for some infectious agents are different from previous method, therefore some test results may need a period of time for comprehensive clinical appreciation.Furthermore,due to the parallel connection of multiple instruments included in the system, more rigorous and frequent quality control becomes a necessity,which may rely on more strict quality control procedure to guarantee the quality.Conclusions The application of the automation system significantly enhanced the efficiency of clinical laboratory all round.In addition, by the quantitative indicators, it is possible to monitor the system operation performance real time, which may feedback and facilitate the improvement constantly,and result in auto confirmation the majority results,eventually.

Objective To compare the clinical effect of 2D and 3D laparoscopic radical prostectomy and summarize surgical experience of laparoscopic radical prostectomy of early prostate cancer.MethodsThe clinical data of 34 cases of prostate cancer treated in our institute from November 2015 to April 2016 were collected and analyzed retrospectively. The patients in observation group (11 cases) were treated by 3D laparoscopic radical prostectomy, while those in control group (23 cases) were given 2D laparoscopic radical prostectomy. The operation time, intraoperative bleeding volume, postoperative drainage time, quantity of drainage fluid within 24 hours postoperatively, indwelling catheter time, hospital time, positive surgical margin rate, potence rate, 30d-urinary continence rate and complications were compared between the two groups.Results All operations were successfully performed. There were no signiifcant differences in operation time, intraoperative bleeding volume, postoperative drainage time, quantity of drainage lfuid within 24 hours postoperatively, indwelling catheter time, hospital time, positive surgical margin rate, potence rate, 30d-urinary continence rate and complications between the two groups (P > 0.05). In observation group, the operation time was (153.52 ± 30.47) min and the potence rate was 50.0 %, 4 cases with uroclepsia (36.4 %), 1 case with urine leakage (9.1 %), no patient had urethral stricture or positive surgical margin, the 30d-urinary continence rate was 72.7 %. In control group, the operation time was (164.73 ± 28.65) min and the potence rate was 38.9 %, 13 cases with uroclepsia (56.5 %), 4 cases with urine leakage (17.4 %), 1 case with urethral stricture (4.3 %), 2 cases with positive surgical margin (8.7 %), 30d-urinary continence rate was 60.9 %.ConclusionLaparoscopic radical prostectomy is a safe, effective and less invasive method for treating early prostate cancer patients. Also 3D laparoscopic radical prostectomy play the similar functional results compared with 2D laparoscopic radical prostectomy, but 3D laparoscopic has the advantage in three dimensions space sense and accurate operation, it is worthy of promoting clinical application.