Objective To explore the effects of the clonidine-displacing substance(CDS)on the function of isolated islets from normal rats. Methods The SD rat islets were isolated by digestive method with collagenase and DNase, and incubated in RPMI 1640 culture medium overnight. Multiscreen Assay System was used to observe the effects of CDS on the function of the isolated islets. Insulin and glucagon were measured by RIA. Results Glucagon secretion of the isolated islets in incubation medium was inhibited by glucose. CDS could significantly inhibit glucagon secretion in concentration-dependent manner. Insulin secretion of the isolated islets was dependent on the glucose concentration of incubation medium. CDS could stimulate insulin secretion significantly. Nifedipine (Ca 2+ channel blocker) and diazoxide (K + channel opener) could inhibit the insulin release of the isolated islets, the effect of which could be abolished by CDS. Conclusion As an endogenous ligand of imidazoline receptors, CDS could stimulate insulin secretion, and inhibit glucagon release in the isolated islets.

AIM:To investigate the inhibition of breviscapine on apoptosis of cultured myocardial cell of neonatal rat induced by hypoxia/reoxygenation. METHODS:Myocardial cell hypoxia/reoxygenation model was established by culturing primary myocardial cells of neonatal rats in vitro. Cultured myocardial cells were divided into 5 groups:control group,hypoxia/reoxygenation group and 3 groups pretreated with breviscapine of final concentration 25,50 and 100 mg/L,respectively. The cell viability was measured with MTT; apoptotic rates were determined by AnnexinV-FITC/PI; the expression of Bcl-2 was detected by immunohistochemical method. Expressions of Cytochrome C (CytC) and Caspase-3 were detected by Western blot. RESULTS:Compared with the control group,the viability of myocardial cell decreased and apoptosis rate elevated after hypoxia/reoxygenation. However after pretreatment with 25,50 and 100 mg/L breviscapine,respectively. Cell viabilities increased and apoptotic rates lowered,and the protective effect on myocardial cell had concentration-dependent. In addition,Expression of Bcl-2 decreased but Caspase-3 activity and CytC release increased in myocardial cells induced hypoxia/reoxygenation. Pretreated with breviscapine,expression of Bcl-2 elevated but Caspase-3 activity and CytC release reduced obviously. CONCLUSION:It is associated with the increase in Bcl-2 expression,inhibition of CytC release and Casepase-3 activity that breviscapine could significantly protect myocardial cell against apoptosis induced by hypoxia/reoxygenation.

Objective:To investigate the clinical and molecular genetic features of neonatal congenital Netherton syndrome (NS) caused by mutations in serine protease inhibitor of Kazal type 5 ( SPINK5) gene. Methods:This study retrospectively analyzed the clinical data of an NS neonate admitted to Shanghai Children's Hospital in November 2018. SPINK5 gene was analyzed using high-throughput sequencing and Sanger sequencing. Relevant articles were retrieved from various databases including China National Knowledge Infrastructure, Wanfang and PubMed, and the reported cases who were diagnosed as NS within two months after birth with SPINK5 gene sequencing results were reviewed. Clinical features, gene mutations, treatment and follow-up results of NS were summarized using descriptive statistical analysis. Results:The patient presented with diffuse erythema and desquamation, sparse hair and repeated infections shortly after birth. Laboratory tests revealed elevated IgE (111 IU/ml) and "invagination-like" change in the hair under optical microscope. SPINK5 gene analysis found that there were compound heterozygous mutations of c.2468dup (p.Lys824Glufs*4) and c.377_378del (p.Tys126*) in the child. The pedigree analysis found that the two mutations were respectively inherited from the father and the mother, which supported the diagnosis of NS caused by SPINK5 gene mutation. Though skin rash improved after comprehensive treatments including anti-infection therapy, gamma globulin injection and skincare, the patient suffered from recurrent infection and was discharged from the hospital after giving up treatment and died of infection at two months old. Eleven NS cases were retrieved from literature and altogether 12 cases were analyzed here. The most common clinical manifestations in the 12 patients were early skin diffuse erythema and desquamation (12/12), infection (8/12), dry hair (7/12), hypernatremia dehydration (7/12), high IgE (5/12), growth retardation (4/12), respiratory failure (3/12), atopic constitution (2/12), diarrhea (2/12), dysphagia (1/12), hypothermia (1/12), wheezing (1/12), hypertension (1/12), liver failure (1/12) and metabolic alkalosis (1/12). Conclusions:NS is caused by SPINK5 gene mutation with atypical manifestations in neonates. Neonates with diffuse erythema and desquamation of the skin, repeated infections, dry hair and especially with high blood IgE should be considered the possibility of NS. Genetic testing is conducive to early diagnosis, guiding treatment decisions and providing a basis for genetic counseling.

ObjectiveTo explore the molecular epidemiological characteristics of group A rotavirus diarrhea in children in Shanghai and to provide the background data for the implementation of rotavirus vaccination.MethodsA total of 910 stool samples were collected from the outpatient children with acute diarrhea from August 2008 to July 2009.Group A rotavirus was detected by usingcommercial colloidal gold device.Rotavirus strains were characterized for G and P genotypes using the nested reverse transcription polymerase chain reaction (RT-PCR).ResultsGroup A rotavirus was detected in 268(29.4％) out of 910 stool samples.Rotavirus infection was found year-round and the peak season was from October 2008 to January 2009,with the detection rates ranging from 38.3 ％ to 70.5％.Ninety-one percent of children (244 cases) with rotavirus-associated diarrhea occurred in children ＜3 years of age.The detection rate of rotavirus was highest (36.6％) in children aged 12-23 months.Among the 268 group A rotavirus-positive strains,G1 was the most common G genotype (65 strains),accounting for 24.3％,followed by G3 (40 strains,14.9％),G mixed genotypes (37strains,13.8 ％),G2 (27 strains,10.1％),G9 (14 strains,5.2％),G4(5 strains,1.9％),other G types (5 strains,1.9％),and unclassified G type (75 strains,28.0％).P[8] and P[4] were the most common P genotypes,accounting for 54.9％ (147 strains) and 11.9％ (32 strains),respectively,followed by P mixed genotypes (6 strains,2.2％) and other P genotypes (4 strains,1.5％),unclassified P type (79 strains,29.5％).The G/P genotype combinations were found as follows:G1P [8] (13.4％),G3P[8] (13.4％),GmixP[8] (10.1％),G1P[4] (8.2％),G9P[8] (2.2％),G2P [4] (1.9％),G1Pmix (1.9％).ConclusionsGroup A rotavirus is a major causative agent of diarrhea in infants and young children in Shanghai.The peak season of rotavirus infection appears in fall and winter.The currently licensed rotavirus vaccines cover the majority of genotypes of rotavirus strains prevailing in Shanghai.

Objective To investigate the expression of Toll like receptor 2 (TLR2) mRNA and TLR4 mRNA in renal tissues of rats with endotoxin-induced shock, and explore the possible effects and mechanism of vasoactive intestinal peptide (VIP) and methylprednisolone ( MP) on renal injury. Methods Endotoxin (E coli O55: B5 LPS, 10 mg/kg) was injected intravenously to the rats to establish the endotoxin-induced shock models ( LPS group, n = 16). The rats in LPS + VIP group (n = 16) was injected with VIP (5 nmol/kg) after model establishment, and those in LPS + VIP + MP group (n = 16) was injected with VIP (5 nmol/kg) and MP (3 mg/kg) after model establishment. Besides, normal saline control group (control group) was established( n = 8). Electron microscope was used to observe the histopathological changes in renal tissues 6 h and 24 h after model establishment, and RT-PCR was performed to evaluate the expression of TLR2 mRNA and TLR4 mRNA in renal tissues. Results Six h after model establishment, glomerular endothelial cell swelling and vacuolar degeneration of mitochondria were observed in LPS group under electronic microscope. Twenty-four h after model establishment, mitochondrial swelling, vacuolar degeneration and slight nucleus pyknosis of renal tubular cells were observed in LPS group. The renal pathological changes in LPS + VIP group and LPS + VIP + MP group were much less severe than those in LPS group at the same time. The expression of TLR2 mRNA and TLR4 mRNA in LPS, LPS + VIP and LPS + VIP + MP groups was significantly higher than that in control group 6 h after LPS injection (P <0.01), while the expression of TLR2 mRNA and TLR4 mRNA in LPS group was higher than that in the other groups 24 h after LPS injection (P < 0.01) . Conclusion The expression of TLR2 mRNA and TLR4 mRNA in renal tissues of rats with endotoxin-induced shock is markedly up-regulated. VIP and MP can obviously alleviate renal pathological changes, and the mechanism may contribute to impacting on the expression of TLR2 mRNA and TLR4 mRNA.

Objective To explore the efficacy and safety of bedside continuous blood purification (CBP) in the treatment of neonatal multiple organ failure (MOF).Methods Totally 6 newborn infants of MOF were hospitalized in department of neonatology in our hospital from June 2011 to June 2013.These 6 cases of clinical data were retrospectively analyzed,6 neonates were treated with CBP combined with conventional treatment.The model for CBP was continuous veno-venous hemodialysis filtration (CVVHDF),blood flow velocity was 3 to 5 ml/(kg· min),replacement fluid dose was 20 to 30 ml/(kg· h),dialysis fluid dose was 15 to 25 rnl/(min· m2).The clinical outcome measures included,blood pressure,blood pH,K+,Na+,blood urea nitrogen,creatinine,urine volume,PaO2/FiO2 and epinephrine intravenous dose,respectively before CBP treatment,6 h,12 h,24 h,48 h after CBP treatment and the end of CBP treatment.The efficacy of CBP treatment was evaluated in neonatal MOF.Results Gestational age of 6 neonates with MOF was 33 to 41 weeks,2 to 19 days old,2.25 to 3.36 kg birth weight.Primary disease was 4 cases of neonatal septicemia(1 case with congenital hereditary metabolic disease),2 cases of severe neonatal asphyxia.All 6 cases of venous catheter were smoothly done.CBP treatment persisted for 49 to 106 hours.Compared with before CVVHDF treatment,blood K+,blood urea nitrogen,creatinine significantly decreased at 12 h after CVVHDF treatment [(5.32 ± 1.84) mmol/L vs.(9.81 ±3.61) mmol/L,(9.0 ±3.4) mmol/L vs.(12.8 ±6.1) mmol/L,(99 ± 16) μmol/L vs.(176 ±25) μmol/L,P ＜0.05],and reached the normal range at 24 h after treatment,urine volume significantly increased at 24 h after treatment (P ＜ 0.05).PaO2/FiO2 reached 200 mmHg (1 mmHg =0.133 kPa) at 6 h after treatment and more than 300 mmHg at 24 h after treatment(P ＜0.05).Fifty percent of epinephrine intravenous dose were down-regulation at 12 h after treatment and stopped using epinephrine at 48 h after treatment.CBP treatment of 6 cases showed effective.Conclusion Application of bedside CBP treatment in neonatal MOF is safe,can effectively help neonates with MOF to skip over renal failure stage.

Objective To investigate the effects of permissive hypercapnia (PHC) strategy used in ventilated newborn swine with respiratory failure on cerebral blood flow (CBF) and tissue metabolism. Method Eighteen newborn swine were randomly (random number) divided into 3 groups (n = 6): PHC, normocapnia (NC) and normal control groups. In PHC (PaCO2 50～60 mmHg) and NC (PaCO2 35 ～ 45 mmHg) groups, newborn swine were ventilated for treating respiratory failure induced by meconium aspiration. CBF was measured by using colored microsphere tracking technique. Cerebral oxygen metabolism rate (CMRO2), cerebral glucose metabolism rate (CMRGlu), and cerebral lactate production (CLP) were measured. Results After ventilation for 6 hours and 12hours, CBF in NC group decreased more significantly than those did in control group, and CMRGlu and CLP increased more significantly than those did in control group. In NC group, CMRO2 decreased more prominently than it did in control group 12 hours later. The CBF/CMRGlu and CBF/CLP ratios in NC group were lower than those in control(P＜0.05). There was no significant difference in CBF/CMRO2 ratio between NC and control groups. After ventilation for 6 hours, CBF in PHC group was lower than those in control group. But after ventilation for 12hours, CBF in PHC group increased and there was no significant difference in CBF between PHC group and control group (P＞O.05). There were no significant differences in CMRGlu, CLP and CMRO2 between PHC group and control group. The CBF/CMRGlu and CBF/CLP ratios in PHC group were lower than those in control, but higher then those in NC group (P＜0.05). There was no significant difference in CBF/CMRO2 ratio between PHC and control groups. Conclusions The reduction of cerebral blood flow and the disturbance of CBF autoregulation disturbance occur in ventilated newborn swine with hypoxemia respiratory failure. PHC may attenuate the reduction in CBF and the disturbance of CBF autoregulation.

B→O blood conversion has very important meanings in the blood transfusion during the period of peace and war.In the leader of Professor Zhang Yangpei,from Academy of Military Medical Sciences,B→O blood conversion has been achieved by using gene engineering.They obtained the great breakthrough in the domain of blood type conversion.In order to realize standardization and automatization,it is necessary to develop a device applying to B→O blood conversion.This device automatically accomplishes the process of B→O blood conversion,shortens the time of B→O blood conversion to quarter than the time by manual operation,and ensures the standardization and obturation.In the condition of one-off pipeline,this device meets the clinical demands for blood,and lays a foundation for clinical expansion of B→O blood conversion.This paper presented the control system of B→O blood conversion device,including design of software and hardware.This device provides a safe and efficient tool for B→O blood conversion.

Objective To investigate the changes of N-terminal pro-brain natriuretic peptide (NT-proBNP) in neonates with hypoxic-ischemic encephalopathy (HIE) complicated by myocardial ischemic injury. Methods Forty neonates with HIE ( 16 cases with concurrent myocardial injury and 24 cases without) were enrolled. Twenty healthy neonates were used as control. Plasma NT-proBNP levels were measured using enzyme immunoassay. Cardiac function was measured by echocardiography. Results ( 1 ) The mean plasma NT-proBNP levels in patients with myocardial injury[(350 ± 56) pmol/L]were significantly higher than those in patients without myocardial injury[(135 ± 37 ) pmol/L]and in the control group [(117 ±23) pmol/L](P ＜0. 05). (2) The NT-proBNP levels in mild,moderate and severe HIE neonates were ( 132 ±34) pmol/L, (247 ±43) pmol/L and (343 ±53) pmol/L. Compared with the control group,the NT-proBNP levels in the neonates with moderate and severe HIE significantly increased. There were significant differences in the NT-proBNP levels among the mild, moderate and severe HIE neonates ( P ＜ 0. 05 ).(3) In patients with myocardial injury,the NT-proBNP levels significantly decreased in the convalescent phase [(250±78) pmol/L]compared with those in the acute phase[(350±56) pmol/L](P ＜0.05). (4) The NT-proBNP levels were significantly related with left ventricular ejection fraction. Conclusion Plasma NT-proBNP levels increase in neonates with HIE complicated with myocardial ischemic injury in the acute phase.Detection of NT-proBNP levels maybe useful in the diagnosis of myocardial ischemic injury and severe HIE.

In order to realize standardization and automatization,it necessary to develop a device for B→O blood conversion.This paper emphasizes on the control system of B→O blood conversion device,including software and hardware design,which provides a safe and rapid method for B→O blood conversion.