BACKGROUND:Transgenosis of basic fibroblast growth factor (bFGF) gene has been successfully performed into the muscle satellite cells of rat extraocular muscles in the previous study of the research group, proving that bFGF could express in the myoblasts of extraocular muscles, also promote cell proliferation and differentiation.OBJECTIVE: To further investigate the methods for regulating the expression of the bFGF in myoblasts following transfection. METHODS: Target gene bFGF was connected with inducing expression vector pcDNA4/T0/myc-His?A, followed by masculine clone sequencing identified by colony PCR and enzyme digestion, EcoR I and Hind III restriction enzyme digestion, as well as Xho I single enzyme verification. C2C12 myoblasts antibiotics sensitivity was screened and finally defined. By use of lipofection transfection technology, cell lines where C2C12 stably expressed pcDNA6/TR were estabolishd and then identified by Western blot. The pcDNA4/TO/myc-His?A-bFGF was transfected into pcDNA6/TR- C2C12 cells. The bFGF expression and secretion in C2C12 cells following tetracycline-induced pcDNA4/TO/myc-His?A-bFGF transfection were determined by immunofluorescence and Western blot, the controls were established.RESULTS AND CONCLUSION: ① The conjunction between the bFGF and inducing expression vector pcDNA4/TO/myc-His?A was proved successfully by sequencing comparison, double digestion and single digestion. ②The minimal lethal concentration of blasticidin to C2C12 cells was 10 mg/L, while that of zeocin was 750 mg/L. ③ The pcDNA6/TR-C2C12 cell lines were established correctly. ④ The myoblasts treated by tetracycline and transfected with pcDNA4/TO/myc-His?A-bFGF were positive for gene expression, those untreated exhibited a negativity; bFGF protein could be produced in myoblasts treated by tetracycline and transfected with pcDNA4/TO/myc-His?A-bFGF, the production reached a peak at 24 hours, while those untreated can not produce bFGF protein. Results suggest that the bFGF expression in the myoblasts can be controlled by tetracycline inhibition and regulatory systems.

Objective To study the repairing effects of nerve growth granule(NGG) on rat common peroneal nerve transection injury.Methods After 50 Sprague-Dawley rats were subjected to nerve suture after transaction,they were randomly divided into 5 groups for daily intragastric administration of drugs:NGG high-dose(5.2g/kg),medium-dose(2.6g/kg),low-dose(1.3g/kg) groups,mecobalamin group(positive control) at 625 ?g/kg,control group(control group control).The drug administration lasted for 4 weeks.Footprint test was performed 2-,3-and 4-weeks after surgery to evaluate toe spread function(TSF).Electrophysiology was performed 4 weeks after operation to determine the compound muscle action potential(CMAP) and nerve action potential(NAP).The number of regenerated myelinated nerve fibers,thickness of myelin sheath and cross sectional area of tibial muscle were measured by histomorphology.Results TSF,amplitude and recovery rate of CMAP and NAP,the number of regenerated myelinated nerve fibers,thickness of myelin sheath and section area of tibial muscle were all increased significantly in a dose-dependent manner compared with the control group.Conclusion NGG contributes to axon growth and myelination,and thus promotes peripheral nerve regeneration in rats with functional recovery.

Objective To assess the risk factors for the occurrence of hypokalemia in patients with hepatic cancer after transcatheter arterial chemoembolizaion (TACE) therapy,and to discuss the corresponding nursing countermeasures.Methods The clinical data of 214 patients with hepatic cancer,who received TACE during the period from August 2014 to February 2015,were retrospectively analyzed.The risk factors causing hypokalemia were analyzed.Results Among the 214 patients,post-TACE hypokalemia occurred in 23 (10.7％).The main risk factors that could cause hypokalemia included anorexia,hydration,vomiting,ascites drainage,sweating.After actively symptomatic treatment,the serum potassium level returned to normal in 22 patients.One patient developed hepatic encephalopathy coma complicated by hepatorenal syndrome,the patient's family members gave up treatment and,according to family members' will the patient left hospital.Conclusion anorexia,vomiting,hydration,ascites drainage,sweating are the risk factors that can cause hypokalemia in patients with hepatic cancer after TACE therapy.The use of low potassium risk scale is helpful for the formulation of nursing countermeasures.

Aim Toinvestigatetheeffectsoftriptonot-erpene methyl ether ( TME ) , a diterpene derived from the medicinal plant Triptergium wilfordii, on human gastric cancer AGS cell proliferation inhibition and ap-optosisinducedinvitro.Methods MTTassaywas used for screening tumor spectrum and detecting the vi-ability of AGS cells and normal human gastric epitheli-al cells GES-1 . Cell morphology was observed by light microscopy and AO / EB staining. Flow cytometry was used to detect cell apoptotic rate and cell cycle. JC-1 staining and fluorescence probe DCFH-DA were em-ployed to detect the changes of mitochondrial mem-brane potential and reactive oxygen species ( ROS ) . The effect of inhibiting AGS clonogenic survival was as-sayed by the method of plate clone formation. Western blot was used to analyse the expression of caspase-3 , caspase-8,Bcl-2andBax.Results MTTresults showed that TME exhibited significantly higher cytotox-icity to gastric cancer AGS cell line than to noncancer-ous cell line GES-1. IC50 for AGS of 48 h treatment was 23 . 85 μmol · L-1 . TME significantly inhibited colony formation and caused morphological changes in AGS cells. Annexin V-FITC / PI double staining showed the apoptotic rate increased. DCFH-DA stai-ning showed TME resulted in an increase in intracellu-lar ROS levels. Mitochondrial membrane potential de-creased after TME treatment. Western blot results showed that TME increased the proportion of Bax /Bcl-2 , with the activation of caspase-8 and caspase-3 . The broad-spectrum caspase inhibitor z-VAD-fmk pre-treatment reduced the expression of caspase-8 and caspase-3. TME enabled AGS cell cycle arrest in G0/G1phase.Conclusion TMEpossessespotenttumor selected toxicity and can induce apoptosis of AGS cells through cell cycle arrest, which is associated with Bcl-2 protein family.

Objective To prepare carcino-embryonic antigen (CEA) targeted and paclitaxel loaded phase-shifting PLGA nanoparticles (Ab-PTX-NPs),and investigate the targeting capability and inhibition to the ovarian cancer cell in vitro.Methods Single-emulsion/solvent evaporation (O/W) and carbodiimide method were used to prepare the Ab-PTX-NPs.The size of nanoparticles was determined by Malvern analyzer.The encapsulation and drug loaded efficiency of paclitaxel were detected by high performance liquid chromatography.And the drug release characteristics was measured by dialysis method in constant temperature shaker.The targeting ability of Ab-PTX-NPs to the ovarian cancer SKOV3 cell was evaluated by the laser scanning confocal microscope and flow cytometry.And the inhibition ability of Ab-NPs was investigated by the CCK-8 assays.Results The size of Ab-PTX-NPs was (397.70±99.95)nm.The encapsulation efficiency and drug loading capacity of PTX were (67.26±4.15) ％ and (6.31±0.39) ％,respectively.The conjugating rate of Anti-CEA antibody was (92.74 ± 5.75) ％.The targeting study in vitro showed that such a number of contrast agents landed around the SKOV3 cells in targeting group,and the mean fluorescence intensity of ovarian cells in targeting group was significantly higher than other groups (P＜0.05).After 24 h,the viability rate of ovarian cells in targeting group was lower than the non-target group (P＜0.05),only higher than that of the pure PTX group (P＜0.05).But there was no significant difference between the targeting group and the pure PTX group (P＞0.05) at 48 h.Conclusion The CEA targeted and paclitaxel loaded phase-shifting PLGA nanoparticles are successfully prepared.It can enhance ultrasound imaging well after activated by LIFU.With high drug-loading efficiency and fast drug release velocity,the Ab-PTX-NPs appeares great targeted ability.

Objective To explore the effects of early hospital-family comprehensive rehabilitation on the development of pretenn infants. Methods A total of 256 premature infants were chosen and divided into an early intervention group (n = 148) and a control group (n =108). Besides being given the conventional mothering instruction , the early intervention group was given early assessment, regular visits and hospital-family comprehensive rehabilitation treatment. The control group was only given the conventional mothering instruction. The fine and gross motor growth quotients, adaptation, verbalisation and social behavior and general growth quotient of all the premature infants were assessed periodically using the infant neuropsychology growth scale. Results The growth quotient assessment indicators of the infants receiving the intervention were obviously better at the 6th, 12th, 18th and 24th month after birth, and the differences between the two groups were statistically significant. The incidence of cerebral palsy in the early intervention group was 0.71% (1/148), with only one cerebral palsy infant in the early intervention group who was at level Ⅲ of the gross motor function classification system ( GMFCS) , while the incidence of cerebral palsy in the control group was 5.1% (5/98) , with 5 cerebral palsy infants, one of whom was at GMFCS level Ⅲ and 4 of whom were at level Ⅳ. Conclusions Early systematic hospital-family comprehensive rehabilitation can improve the general growth of premature infants, decrease the incidence of cerebral palsy, and neurobehavior deficits.

Objective To explore the role of clinical pharmacists in rational drug use through the pharmacy care of an elderly pneumonia patient with Chlamydia psittaci infection and drug-induced liver injury. Methods The clinical pharmacists participated in the treatment of one patient with Chlamydia psittaci pneumonia and drug-induced liver injury. Based on the results of second-generation gene sequencing, the characteristics of the pathogen were learned by literature search. The clinical pharmacists monitored the patient’s liver and kidney function, provided a new medication treatment plan to Doctors, and performed patient education during the treatment. Results The initial empirical anti-infective treatment with teicoplanin and imipenem-cilastatin was not effective. After the diagnosis of Chlamydia psittaci and Candida albicans infection, the combination of doxycycline with azithromycin and fluconazole was administered. Drug-induced liver injury was found with this treatment. The clinical pharmacist proposed to switch to doxycycline and clarithromycin with co-administration of magnesium isoglycyrrhizinate and polyene phosphatidylcholine to protect the liver. With this new regime, patient's liver function was improved and the infection was under control. Conclusion Individualized pharmaceutical cares provided by clinical pharmacists helped the safe, rational and effective use of medications.

[ ABSTRACT] AIM:To investigate the preventive effects of Clostridium butyricum ( C.butyricum) on the type of pylorus ligated gastric ulcer ( GU) in mice and the underlying mechanisms.METHODS:ICR mice were randomly divided into 4 groups:sham operation group, model group, C.butyricum pretreatment group and omeprazole pretreatment group. Gastric pyloric ligation was adopted to establish GU model in mice.The gastric juice was collected to measure the content of gastric free mucus, the pH of gastric juice and the activity of pepsin.The gastric tissues were collected for routine HE stai-ning to observe the pathological changes.The content of glycogen was detected by PAS staining.The protein expression of Bax and Bcl-2 in the gastric mucosa was also assessed by immunohistochemical staining.RESULTS: The HE and PAS staining showed that the C.butyricum pretreatment obviously attenuated the mucosa lesion induced by ligation.Compared with model group, the pH of gastric juice was significantly raised.The activity of pepsin fell off in C.butyricum group, which was lower than that in omeprazole group.In comparison with model group, the content of gastric free mucus was dra-matically increased and PAS staining showed a significant rise in C.butyricum group, but not in omeprazole group.The protein expression of Bax was decreased and the protein expression of Bcl-2 was upgraded in C.butyricum group than those in model group.CONCLUSION:C.butyricum protects gastric mucosa against the challenge of pylorus ligation in mice and its mechanism may be related to inhibiting gastric acid secretion and the activation of pepsin, increasing the production of gastric free mucus, strengthening the expression of bcl-2 gene and inhibiting the expression of bax gene.

Objective: : This study analyzed the influence of p120-catenin (catenin [cadherin-associated protein], delta 1 [CTNND1]) on the malignant characteristics of glioma and elucidated the potential underlying mechanism. Methods: : The p120 expression level was assessed in the brain tissues of 42 glioma patients and 10 patients with epilepsy by using the immunohistochemical method. Meanwhile, quantitative polymerase chain reaction (QT-PCR) technology was employed to assess the expression of p120 in the brain tissues of 71 glioma patients and 13 epilepsy patients. LN229, U251, and U87 glioma cells were used for in vitro analysis and categorized into four treatment groups : siRNA-blank control (BC) group (no RNA sequence was transfected), siRNA-negative control (NC) group (transfected control RNA sequences with no effect), and siRNA-1 and siRNA-2 groups (two p120-specific interfering RNA transfection). p120 expression in these treatment groups was quantified by western blotting assay. The migratory and invasive capabilities of glioma cells were studied by wound healing assay and Transwell invasion assay, respectively, under different treatment conditions. MTT (3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide) assay and cell cycle and apoptosis assay were used to determine glioma cell proliferation and apoptosis, respectively. Enzymelabeled assay was performed to measure intracellular calcium ion concentration. Immunofluorescence assay was performed for determining microtubule formation and glioma cell distribution. Results: : Brain tissues of the glioma group exhibited a remarkable increase in the p120 expression level as compared to brain tissues of the nontumor group (p<0.05). Furthermore, a strong positive correlation was noted between the malignancy degree in glioma brain tissues and p120 expression in Western blotting (r=0.906, p<0.0001) and QT-PCR (F=830.6, p<0.01). Compared to the BC and NC groups, the siRNA transfection groups showed a significant suppression in p120 expression in glioma cells (p<0.05), with a marked attenuation in the invasive, migratory, and proliferative capabilities of glioma cells as well as an increase in apoptotic potential (p<0.05). Enzyme-labeled assay showed a remarkable increase in calcium concentration in glioma cells after siRNA treatment. Immunofluorescence assay revealed that the microtubule formation ability of glioma cells reduced after siRNA treatment. Conclusion : p120 has a pivotal involvement in facilitating glioma cell invasion and proliferation by potentially modulating these processes through its involvement in microtubule formation and regulation of intracellular calcium ion levels.

OBJECTIVETo identify the mortality and epidemiological pattern of dementia and its various major subtypes among urban and rural senior residents in Beijing.

METHODSBased on Beijing's dementia prevalence survey among residents aged 55 years and above in 1997, respondents were selected by stratified multiple-stage cluster sampling and 12 urban communities and 17 rural village communities were randomly sampled then follow-up in 2001. COX regression was used to analyze relative risks controlling confounding factors on deaths of dementia cases.

RESULTSThe mortality of dement patients in the 55-64 age-group was 0.82/100 person-year. The age-standardized mortality of dement cases was 0.90/100 person-year. The mortality in the 65 and above age-group was 1.44/100 person-year, and the age-standardized mortality was 1.56/100 person-year. Among AD cases, the above two mortalities were 0.35/100 and 0.42/100 person-year respectively, and among VaD cases, 0.34/100 and 0.36/100 person-year respectively. For both AD and VaD cases, their mortality increased with age. Region, gender and age were more significant to survival of AD cases.

CONCLUSIONOne major subtype of dementia, AD, among elderly urban and rural residents in Beijing, has a different mortality and epidemiological pattern from VaD.

Aged ; Aged, 80 and over ; Cardiovascular Diseases ; mortality ; Cause of Death ; China ; epidemiology ; Dementia ; mortality ; Female ; Humans ; Male ; Middle Aged ; Neoplasms ; mortality ; Prevalence ; Respiratory Tract Diseases ; mortality ; Rural Population ; statistics & numerical data ; Urban Population ; statistics & numerical data