BACKGROUND:Previous studies have demonstrated that transplanted neural stem cells can survive and proliferate in the brain of Alzheimer disease(AD)rats,however,it is poorly understood whether it can rebuild the nerve tracts by substituting the injured or dead neurons and improve learning and memory abilities.Synaptophysin is one of the important markers of synaptic rebuilding.OBJECTIVE:To observe the effects of neural stem cell transplantation on synaptophysin expression in hippocampus and learning and memory abilities of AD rats.METHODS:Sprague Dawley rats were randomly divided into the normal control,AD model,2-week-transplantation and 4-week-transplantation groups.All rats were established AD models except that in the normal control group.Neural stem cells were isolated from the dentate gyrus of hippocampus of newborn rats,labeled with Hoechst33258,and then transplanted into CA1 region of hippocampus of rats in the 2-week-transplantation and 4-week-transplantation groups.The behavioral testing in the rats was performed using Y-maze trial.Nissl staining and synaptophysin immunohistochemistry were detected after the rats were sacrificed.The same volume of stroke-physiological saline solution was injected into rats in the AD models group using the identical methods.There was no treatment in the normal control group.RESULTS AND CONCLUSION:①The cells number in the hippocampal CA1 region of the 2-week-transplantation and 4-week-transplantation groups were increased than that of AD model group,but were still less than that of the normal control group(P ＜ 0.05).There was no significantly difference between the absorbance values of 2-or 4-week-transplantation group and control group(P ＞ 0.05).②The absorbance values of the 2-week-transplantation and 4-week-transplantation were significantly greater than that of the control and AD model groups(P ＜ 0.05).③The learning and memory abilities in 2-and 4-week-transplantation group enhanced obviously and their correct reaction rates improved evidently,which was found statistically significant difference from AD model group(P ＜ 0.05),while no statistically significant difference from control group(P ＞ 0.05).The transplanted neural stem cells may promote the synaptic rebuilding and improve learning and memory abilities in AD rats.

Objective To explore the long-term effect, prognosis and administration of corticosteroid treatment on autoimmune pancreatitis (AIP). Methods Clinical data were analyzed in 13 diagnosed and followed up AIP patients of Peking Union Medicine College Hospital during August 2004 to August 2008. Results Of 13 patients, 12 were males and 1 was female, with a mean age of 58.7 years old, and a mean follow-up of 30 months. Of 11 patients compliated with bile duct disease,biliary stents were placed in 9 patients and already taken out. Corticosteroid treatment was received by cured patients. The average corticosteroid therapeutic time was 9.2 months, 7.9 months in 6 biliary stent placed patients, 13.4 months in corticosteroid treated alone patients, the statistical difference was significant (P = 0. 023). Serum inflammatory parameters normalized range from 5. 3 to 8.8 weeks. After corticosteroid treatment, pancreas enlargement improved in all patients at the first imaging reexamination (1.0 to 11.3 weeks), pancreatic size normalized in 9 patients with an average of 16.6 weeks corticosteroid treatment. No relapsing sign was found with imaging examination during follow-up. Of 8 newly onset diabetes patients, glucose level normalized in 4 patients after corticosteroid treatment. Two patients complicated with autoimmune hepatitis developed early hepatic cirrhosis symptoms at the end of the follow-up. Swollen submandibular gland enlargement relapsed in one patient after corticosteroid withdrawn for six months. Conclsion AIP patients responsed well to corticosteroid treatment. Placement of biliary stent could shorten corticosteroid therapeutic time.Patients with bile duct complications and newly onset diabetes could partially relieve after the corticosteroid treatment, the prognosis of patients with autoimmune hepatitis complications was relatively poor.

The hepatocellular carcinoma (HCC) is one of the most common malignant tumors.Patients with hepatobiliary cancer suffer from multiple symptoms,such as physical symptoms and psychology symptoms.In recent years,along with the further research of symptoms,the concept of symptom clusters occurred.Those two issues have always been the hot topics in recent years.Through the efforts of experts at home and abroad,there has been a part of progress in this area.However,the results are not the same.There are lots of disputes in the numbers、sequence、incidences of the symptom clusters.The paper will discuss this issue from symptom experience and symptom clusters in order to provide the reference and basis for symptom management.

Objective To investigate the molecular mechanism of bone marrow mesenchymal stem cells (BM-MSCs) in repairing cisplatin-induced acute renal injury.Methods The rats were injected 6 mg/kg of cisplatin intraperitoneally,and bone marrow mesenchymal stem cells (BM-MSCs group) or PBS (PBS group) were injected respectively via tail vein after 24 hours.The rats without injecting cisplatin were selected as a normal control group.The repair effect of BM-MSCs on renal injury was observed by HE staining and immunohistochemistry.In addition,NRK-52E cells were cultured in vitro and treated with cisplatin for 6 hours.Then,NRK-52E cells were continued to culture for 48 hours or co-cultured with BM-MSCs for 48 hours,and NRK-52E cells untreated with cisplatin were used as a control.The expression levels of miR-92b and its target gene PTEN were detected by qRT-PCR,and the expression level of p-Akt by western blot.Results HE staining showed that the tubular protein casts in BM-MSCs group were significantly less than that in PBS group,and that the renal tubular structure was significantly improved in BM-MSCs group.Immunohistochemical staining indicated that the number of cells expressing proliferating cell nuclear antigen (PCNA) in BM-MSCs group (131.0 ± 14.4) was significantly higher than that in PBS group (42.2 ±6.1,t =11.28,P ＜0.01).qRT-PCR results showed that in the vivo experiment,compared with the expression level of miR-92b and PTEN in the normal control group (1.11 ± 0.78,1.01 ± 0.21),PBS group were (4.64 ± 1.06) and (0.61 ± 0.2),respectively (all P ＜ 0.05);BM-MSCs group were (2.27 ± 0.81) and (1.1 ± 0.1),respectively (all P ＜ 0.05).In vitro experiment,compared with the expression level of miR-92b and PTEN in the negative control group (1.12 ± 0.77,1.02 ± 0.13),cisplatin group were (7.64 ± 0.72) and (0.58 ± 0.2),respectively (all P ＜ 0.05),cell group were (4.38 ± 0.50) and (1.15 ± 0.23),respectively (all P ＜ 0.05).Western blot results showed that compared with the expression level of p-Akt in cisplatin group (0.96 ± 0.18),p-Akt expression in cell group was (2.11 ± 0.11,P ＜ 0.01).Conclusion BM-MSCs may repair the cisplatin-induced acute renal injury via down-regulating the expression level of miR-92b.

Objective To explore the changes of plasma brain natriuretic peptide(BNP) and N-terminal pro-B-type natriuretic peptide (NT-proBNP)of sepsis combined with myocardial injury in newborns.Methods According to neonatal sepsis treatment program,45 cases of sepsis newborns in NICU of the Second Hospital in Lanzhou University from Jul.2007 to Jun.2008 were collected.According to the myocardial injury diagnostic criteria,45 cases neonatal sepsis were divided into myocardial injury group(n=22) and non-myocardial injury group(n=23).Myocardial injury group was also divided into congenital heart disease group and non-congenital heart disease group accor-ding to echocardiography.At the same time,30 healthy newborns were collected as healthy control group.Every newborns were tested the level of plasma BNP,NT-proBNP,creatine kinase-MB(CK-MB) and cardiac-troponin I (cTnI).Results There were significant difference between myocardial injury group,non-myocardial injury group and healthy control group in the levels of plasma BNP,NT-proBNP,CK-MB and cTnI,those in congenital heart disease group were higher than those in non-myocardial injury group and the healthy control group(Pa0.05).Conclusions BNP and NT-proBNP can be early used to diagnose myocardial injury and heart failure of neonatal sepsis associated with CK-MB and cTnI.In NICU,infants with sepsis should normally test BNP and NT-proBNP in order to early diagnose myocardial injury of neonatal sepsis.

OBJECTIVE:To establish an HPLC method for the determination of the content of L-arginine in cefradine for injection.METHODS:The chromatographic analysis was carried on LiChrospher Diol column,and the mobile phase was composed of 0.01 mol?L-1 ammonium dihydrogen phosphate buffer solution(adjusted pH to 2.0?0.1 with phosphoric acid)-acetonitrile(25∶75)at a flow rate of 1.0 mL?min-1;the column temperature was 30 ℃;the detection wavelength was 206 nm,and the sample size was 20 ?L.RESULTS:The linear range of L-arginine was 51.2～307.2 ?g?mL-1(r=0.999 9).The average recovery was 100.7%(RSD=0.6%).CONCLUSION:The method is rapid,accurate and simple,and suitable for the determination of L-arginine in cefradine for injection.

Objective To evaluate the safety and clinical efficacy of renal artery stent treatment for severe atherosclerotic renal artery stenosis (ARAS) in the elderly. Methods In a prospective nonrandomized study in our hospital from January 2003 to April 2008, 147 consecutive elderly patients with ARAS (diameter reduction ≥ 65％) underwent percutaneous transluminal renal angioplasty and stenting (PTRAS) for resistant hypertension or reserving renal function. They were followed up for 6- 66 months and the effects of the procedure on renal function, blood pressure and cardiovascular events were observed. Results The success rate of PTRAS was 100％. During 6-66 months of follow up, both systolic and diastolic blood pressure were significantly decreased, and less antihypertensive medication was taken (P<0.01). Serum creatinine was significantly decreased during 6-48 months of follow up (P<0. 05) and did not change significantly during 54-66 months of follow up (P>0. 05). Blood urea nitrogen was significantly decreased during 6-24 months of follow up (P<0.05) and did not change significantly during 30-66 months of follow up (P>0.05). Complications related with the procedure occurred in 7 cases (4.8％). 17 patients failed to follow up (11.6％) and in 18 cases cardiovascular events occurred (12.2％), including 4 cases of renal events (2. 7％), 4 cases of myocardial infarction (2.7％), 2 cases of stroke (1.4％) and 8 cases of cerebral and cardiovascular events (5.4％) during 6-66 months of follow up. The survival rates of free-of-events at year 1, 2, 3, 4 and5 were 91.0%(121/133), 90.2%(83/92), 78.5%(51/65), 73.8%(31/42) and 54.8%(17/ 31), respectively. The survival rates at year 1, 2, 3, 4 and 5 were 94.0% (125/133), 90. 2% (83/ 92), 84.6%(55/65), 73. 8%(31/42) and 61.3%(19/31), respectively. Conclusions Renal artery stent as treatment for ARAS in the elderly has a beneficial effect on blood pressure control and on renal function during middle and long term follow up. The treatment may be helpful in reduction of cardiovascular events and mortality, which should be investigated further.

Objective To obtain highly purified astrocytes and identify the cells in each stage to support further studies.Methods The cerebral cortex of a neonatal SD rat was isolated and prepared into single cell suspension.The obtained cells were purified by differential adherence and shook at a constant temperature.By inverted phase contrast microscopy and HE staining,cell morphology was observed.The immunofluorescence staining with anti-mouse GFAP was used to identify the cells.Results The primary cortical cells developed rapidly at 3 d after culture and covered the flasks at 9-12 d.At this time,the cells showed stratification and the astrocytes lay at the lower layer.GFAP positive rate was only about (67.2 ±7.1)%.After the first passage,GFAP positive rate increased obviously (84.0±6.0)%. However, oligodendrocytes and microglias could not be removed completely,and the cells also showed stratification.Through 3 times of passages,we obtained many single species of astrocytes showing satellite shape with 2 or 3 processes,big cell body and round or oval-shaped nuclei leaned to one side.Immunofluorescence staining showed that nearly all of the cells were strong positive and the positive rate reached as high as (97.6 ± 2.4 )%.Conclusion Through differential adherence and shaking at a constant temperature,more astrocytes of high purity and in good state can be obtained.

Objective To explore the genetic etiologies in 2 siblings with different epileptic encephalopathies (EEs) diagnosed as Ohtahara syndrome(OS) and atypical benign partial epilepsy(ABPE) from one family.Methods The 2 brothers were diagnosed at the Pediatric Neurological Clinic of Peking University First Hospital in September 2013,whose clinical data were collected.The coding region of the syntaxin-binding protein 1 gene (STXBP1) and glutamate receptor subunit gene (GRIN2A) were detected by using Sanger sequencing in the 2 siblings.For the elder brother,targeted next-generation sequencing was further performed to detect the genes associated with epilepsy.Results The younger brother manifested focal motor seizures and tonic spasms in cluster at the age of 1 month.Interictal electroencephalogram (EEG) showed suppression-burst pattern.He was diagnosed as OS.The elder brother had seizure onset at age of 6 years old.Focal motor seizure during sleep was his seizure type.His EEG showed interictal discharges in Rolandic area primarily.Electrical status epilepticus during sleep,epileptic negative myoclonus and intellectual disabilities occurred during the course.He was diagnosed as ABPE.Brain magnetic resonance imagines for both of them were normal.Screening of STXBP1 mutations for the younger brother found a de novo heterozygous mutation:c.1672C ＞ T (p.Q558X).Gene detection for the elder brother and the parents showed negative results.Conclusions Coexistence of distinct EEs was reported in 2 brother siblings:the younger brother had OS associated with a novel nonsense mutation in STXBP1,and the elder brother had ABPE without genetic evidence.This study indicated that different pathological mechanisms might exist underlying the two different EEs in a family.