BACKGROUND: Puerarin possesses various biological efficacies, such as the protective efficacy on hypertension, cardiac disease, diabetes mellitus and blood disease, and the extracts of puerarin can inhibit the proliferation of S180 sarcoma and Lewis lung cancer to some extent.OBJECTIVE: To investigate the modulation effects of total flavone of puerarin (TFP) on the growth of pheochromocytoma PC12 cells and the protective efficacy on the H2O2-induced cellular oxidative damage.DESIGN: Complete randomization design and control experiment.SETTING: Department of Biochemistry and Institute of Geriatrics, Chinese PLA General Hospital.MATERIALS: Puerarin was bought from Tongrentang drug store, and TFP was extracted and purified routinely by ethanol and ether acetate, then was evaluated with thin-layer chromatography. The content of puerarin in extracts was 31.79% in quantitative assay. Methyl thiazolyl tetrazolium (MTT), luminal and anti-oxidative activity reagent xanthine oxidase were all from Sigma Company. PC12 cells were given by Institute of Geriatrics,Chinese PLA General Hospital as a present.METHODS: The experiment was conducted at the Institute of Geriatrics, Chinese PLA General Hospital from January to July in·2001.① The cells were cultured in 20 g/L DMEM (pH 7.1-7.2), and randomized into two groups: TFP group and H2O2-injury+TFP group, and each group was divided into 5 mass concentrations (0, 1.0, 10, 100 mg/L and 1.0 g/L). There were 8 holes for parallel culture with 100 mL culture medium in each hole (containing 1 ×l09 L-1 cells). TFP group:TFP was added for 72-hour culture at 37 ℃; H2O2 injury+TFP group:TFP was firstly cultured for 48 hours at 37 ℃, then 500 mmol/L H2O2 was added and co-cultured for other 24 hours.②The activity of cultured PC12 cells was monitored by MTF assays, the content of nitrite was measured by Griess reagents, and the antioxidant activity of superoxidedismutase (SOD) was monitored by hypoxanthine/xanthine oxidase.H2O2-initiated PC12 cellular oxidative damage had been used as experimental model to study the protective efficacy of TFP, and expressed as inhibition ratio [(blank A value-detection A value)/blank A value × 100%]. The higher inhibition ratio indicated the strong ability of clearing O2-. ③ One-factor analysis of variance was used to compare the difference between data. MAIN OUTCOME MEASURES: The influence of TFP on the nitrite content, SOD activity and cell activity in PC12 cells.RESULTS: ①Effect of TFP on PC12 cell activity: 1-10 mg/L TFP hadno obvious effects on the growth of PC12 cells, and 100 mg/L TFP in creased the cell growth (P ＜ 0.05), whereas the TFP concentration was increased to 1.0 g/L, the activity of PC12 cells was inhibited obviously (P ＜ 0.01). TFP of 1-100 mg/L could protect the cultured cells from the oxidative damages of H2O2 concentration dependently (P ＜ 0.05).② Effect of TFP on clearing O2-: The ability of clearing O2 increased withthe mass concentrations of TFP in both groups with obvious dose-effect relation, except when 1 mg/L TFP was added in the H2O2 injury+TFP group. The SOD activity in PC12 cell culture liquid was obviously en hanced after adding 100 mg/L and 1 g/L TFP, compared with that without TFP addition (P ＜ 0.05-0.01). ③Modulation of TFP on nitrite: TFP of low concentration (1-100 mg/L) reduced the production of cellnitrite, whereas increased the nitrite production at the concentration of1.0 mg/mL.CONCLUSION: ①TFP can regulate the growth of PC12 cells, which canbe enhanced by low-concentration (1-100 mg/L) TFP whereas inhibited byhigh-concentration (1 g/L) TFP. However, the anti-oxidation of TFP is themost powerful. ②TFP can protect the PC12 cells obviously from the oxidative damages induced by H2O2 at low concentration.

Helicobacter pylori (H.pylori) infection rate is increasing these years.The incidence of related diseases subsequently rose.Hp infection can not only cause digestive diseases,but also lead to idiopathic thrombocytopenic purpura (ITP),iron deficiency anemia,and indigestion etc.which can affect the growth and nutritional health of children.H.pylori has become a global issue,both international and domestic colleagues have reached a consensus on treatment for adults.According to the new conscnsus,the aim of this article is to study the diagnosis and treatment options for children with Hp infection.

Objective To investigate the cerebral protection of dexmedetomidine in patients with heart valve replacement under CPB.Methods Totally 80 cases of patients with heart valve replacement under CPB in Seventh people's hospital of Zhengzhou city from January 2015 to December 2015 were selected and divided into observation group and control group,40 cases in each group.Patients in observation group were treated with dexmedetomidine before anesthetic induction and during the operation,and patients in control group were treated with same dose of saline solution.The arterio-venous oxygen content difference (Da-jvO2),jugular venous oxygen saturation (SjvO2),cerebral oxygen extraction rate (CERO2),serum myelin basic protein (MBP),serum neuron-specific enolase (NSE),and S1003 protein content before anesthetic induction (T0) and when the ascending aorta open (T1),After 10 min of CPB (T2) and postoperative 6 h (T3) were compared between two groups,and occurring rates of MMSE and POCD were also compared.Results SjvO2 values of two groups at T1 were significant higher than T0 (P ＜ 0.05),Da-jvO2,and CERO2 were significant lower than T0 (P ＜ 0.05),SjvO2 values of observation group at T1 and T2 were significant higher than those of control group and Da-jvO2 and CERO2 were significant lower than those of control group (P ＜ 0.05);At T2 and T3,MBP levels of observation group and control group were significant higher than T0 (P ＜ 0.05),and those in observation group were significant lower than control group (P ＜ 0.05);At T1,T2,and T3 of observation group and control group were significant higher than T0 (P ＜0.05),and NSE levels of observation group at T1,T2,T3 were significant lower than those of control group (P ＜ 0.05),S-100β levels of observation group at T2 and T3 were significant lower than those of control group (P ＜ 0.05).After 7 d,MMSE score of observation group was significant higher than that of control group and POCD occurring rate was lower (P ＜ 0.05).Conclusion Dexmedetornidine can improve cerebral ischemia hypoxia state and reduce brain damage in patients with heart valve replacement under CPB.

Objective To explore the relative signal transduction pathway of angelica polysaccharide(APS) inducing K562 cells toward erythroid differentiation. Methods K562 cells were divided into control group and ASP group, the control group cells were routinely cultured and APS group cells were treated with APS(final concentration is 100-500mg/L). By using of benzidine staining and spectrophotometry, the characteristics of erythroid differentiation of K562 cell induced by APS were detected;By using of laser confocal microscopy, the distribution of JAK_2 and STAT_5 in K562 cells was observed;By using of Western blotting, the expression of JAK_2 and STAT_5 in nucleus and cytoplasm of K562 cells was detected, By using of imm～uno～pre～cipi～ta～tion, the phosphorylation change of JAK_2 in cytoplasm was tested. Results After being induced by APS, the benzidine staining positive rate of K562 was increased.With increasing the concentration of APS, hemoglobin synthesis in K562 cells was promoted accordingly. After being cultured for 12 hours, 24 hours and 48 hour, the expression of STAT_5 in nucleus of K562 cells induced by APS was significantly higher than that of control group, however, expression of STAT_5 in cytoplasm of K562 cell induced by APS was obviously lower. The expression of JAK_2 in K562 cells was not different between APS group and control group, but the JAK_2 phosphorylation level of APS group was much higher than that of control group.Conclusion APS can induce erythroid differentiation of K562 cells, and the mechanisms may be that APS can promote the phosphorylationthe of JAK_2, then stimulating nuclear translocation of STAT_5.

Objective To explore the clinical features and gene mutations of blepharophimosis-ptosis-intellectual-disability syndrome (BPID). Methods The clinical data, diagnosis and treatment of a child with BPID in neonatal intensive care unit (NICU) were reviewed. Based on the literature retrieved from PubMed database, the common classification, clinical features, diagnosis and genetic counseling of BPID and its affiliated blepharophimosis-mental retardation syndromes (BMR) were reviewed. Results This male infant was 39 weeks of gestational age with birth weight of 1920 g, and was admitted to NICU 15 min after birth due to dyspnea. The main clinical manifestations were facial deformity such as biepharophimosis, ptosis and micromandible, inspiratory dyspnea with laryngeal cartilage softening, malformations of the thorax and feeding difficulties. A heterozygous mutation in UBE3B gene was identified by complete exon sequencing and he was diagnosed of BPID, a rare genetic disorder. Reviewing the literature, there was no relevant report in domestic. While one foreign literature was found to report 5 patients from 4 families having a subtype of BMR, a kind of autosomal recessive diseases caused by mutations in the UBE3B gene. Conclusion BPID is a rare clinical entity of BMR. Complete exon sequencing can be used to diagnose the disease.

ABSTRACT:Objective To explore cardiac function and myocardial collagen type I in diet-induced insulin-resistant rats and the effect of telmisartan on cardiac diastolic function in diet-induced insulin-resistant rats. Methods We randomized 27 Wistar rats into control group (n = 9 ),high-fat group (n = 9 ),and telmisartan treatment group (n = 9 ).At the end of the study,left ventricular end diastolic pressure (LVEDP)and left ventricular systolic pressure (LVSP)of the rats and ± dp/dt were detected by carotid artery intubation.Masson cardiac staining was used to observe cardiac fibrosis,and collagen volume fraction (CVF)was measured.ELISA method was used to detect the concentration of plasma PICP and ICTP.Results Compared with the control group,in high-fat group LVEDP was significantly higher and -dP/dtmax decreased significantly (P < 0.01 );the plasma PICP level and the ratio of PICP/ICTP were significantly increased (P <0.01),cardiac collagen volume
fraction was significantly higher (P <0.01).After 22 weeks’telmisartan intervention,compared with the high-fat group,LVEDP and LVSP were significantly decreased (P < 0.01 ),but -dP/dtmax significantly increased (P <0.05).The level of the plasma PICP and PICP/ICTP were significantly decreased (P < 0.05 );left ventricular myocardial tissue collagen volume fraction content was decreased (P <0.01).The correlation analysis showed that cardiac collagen volume fraction in insulin-resistant group was positively correlated with insulin resistance index but negatively correlated with -dp/dtmax (P < 0.01 ).Conclusion Insulin resistance promoted the synthesis of myocardial type I collagen,leading to increased myocardial collagen deposition and decreased cardiac diastolic function.Telmisartan may improve diastolic function partly by improving insulin resistance and reducing the deposition of myocardial collagen type I.

Objective To investigate cellular and behavioral effects of adenosine A2A receptor antagonist in a rat model of levodopa-induced motor complications.Methods The hemi-parkinsonian rat model was produced by stereotaxically injecting 6-OHDA to right medial forebmin bundle(MFB).Animals were intraperitoneally treated with levodopa 50 mg/kg plus benserazide 12.5 mg/kg twice a day for 22 days levodopa + vehicle.Rotational duration was estimated.After they were sacrificed,the expression of adenosine A2A receptor was observed by immunohistochemistry and Western blot.Results CSC,reversing the shortened rotational duration induced by levodopa,prolonged the rotational duration.This effect was maintained fil the end of the treatment.The chronic levedopa treatment induced an upregulation of adenosine A2A receptor expression in the lesioned striatum (IOD,(11.55±2.75)×104).The subsequent CSC treatment decreased the adenosine A2A receptor expression to the level of control (IOD,(6.02±1.29)× 104) and PD group (IOD,(5.60±1.83)×104>,F=33.31,P<0.05).Conclusion These results suggest that adenosine A2A receptor is probably involved in the development of levodopa induced motor complications and adenosine A2A receptor antagonist could be useful in the treatment of motor comphcations in parkinsonian patients.

Objective:To investigate the effects of electroacupuncture (EA) at Neiguan (PC 6) and Baihui (GV 20) by observing the changes of CCAAT/enhancer-binding protein(C/EBP) homologous protein (CHOP) and caspase-12 gene expressions in rats after cerebral ischemia-reperfusion injury (IRI), and explore whether the apoptosis pathway of endoplasmic reticulum stress (ERS) is involved in the protective mechanisms of EA.
Methods:Sixty rats were randomly assigned to five groups (12 in each group): a normal control group (group A), a sham-operation group (group B), an operation group (group C), an Edaravone group (group D) and an EA group (group E). The cerebral IRI rat model was induced by middle cerebral artery occlusion (MCAO) using intraluminal monofilament. 2,3,5-triphenyl tetrazolium chloride (TTC) staining was adopted in the measurement of cerebral infarction volume. Real-time polymerase chain reaction (RT-PCR) was used to determine the mRNA expressions of CHOP and caspase-12.
Results: Compared with group A and group B, the volume of cerebral infarction and mRNA expressions of CHOP and caspase-12 in group C, group D and group E were increased, with statistical significances (P<0.05 orP<0.01); compared with group C, the volume of cerebral infarction and mRNA expressions of CHOP and caspase-12 in group D and group E were decreased significantly (P<0.05 orP<0.01); there were no significant differences between group D and group E in comparing the above items (P>0.05).
Conclusion:EA at Neiguan (PC 6) and Baihui (GV 20) can effectively suppress the volume of cerebral infarction. Furthermore, the underlying mechanism of EA at Neiguan (PC 6) and Baihui (GV 20) is possibly related to the down-regulation of CHOP and caspase-12 mRNA expressions, so as to decrease cell apoptosis.

Objective To investigate the effect of integrated swallowing function rehabilitation training on nasal feeding patients with pseudobulbar palsy or bulbar palsy after acute ischemic stroke. Methods A total of 54 patients met the diagnosis criteria were enrolled prospectively. They were divided into either an integrated group (n=31)or a control group (n=23)using a random number table method. The patients of the control group received routine treatment of stroke,including brain protection therapy,improvement of cerebral circulation,dehydration,regulating blood pressure,and symptomatic treatment,etc;in addition to conventional treatment,those of the integrated group also received integrated swallowing function rehabilitation training,including neuromuscular electrical stimulation plus swallowing reflex facilitation technique,and oral facial glossopharyngeal function training,etc. The oral intake function in patients of the integration and pseudobulbar palsy or bulbar palsy (according to MRI and signs)was compared between the two groups. The evaluation index was the Functional Oral Intake Scale (FOIS)score. Results (1)The FOIS score (median 4)of the integrated group was higher than that of the control group (median 2). There was significant difference between the two groups (P<0. 01). There was also significant difference before and after comparison between the two groups. (2)The FOIS score of pseudobulbar palsy in the integrated group was higher than that of the control group (median scores 4 and 2 ). There was significant difference (P<0.01);There was no significant difference in the FOIS scores of bulbar palsy between the two groups,but the FOIS score of the integrated group was higher than that of the control group (median scores 4 and 3). (3)The median differences between the integrated group and the control group were 2 and 1 respectively. The increased amplitude of FOIS of the integrated group was higher than that of the control group. There was significant difference (P<0. 01). Conclusion The integrated swallowing function rehabilitation training can improve the oral intake function,significantly reduce irritating cough,increase the capacity of oral intake,and increase the types of food. The effect of the improvement of pseudobulbar palsy is more significant in the integrated group.

Objective To observe and evaluate the application of case-based learning in occupational health teaching of clinical medicine students.Methods Seven-year undergraduates majored in clinical medicine were selected as study subjects,divided into two groups,and received case-based learning(55 persons,story case for theory course,field case for experiment course)and traditional teaching(56 persons,theoretical instruction for theory course,instructional video for experiment course),respectively.T test and x2 test were used to analyze and compare students' after-school evaluation results and the questionnaire survey results of both groups respectively.Results The total score of the after-school evaluation[(11.84 ± 2.86) vs.(9.80 ± 3.39),t=-3.419],basic cognitive ability score[(4.80 ± 1.08) vs.(4.05 ± 1.43),t=-3.097] and analyzing ability score[(7.04 ±2.21) vs.(5.75 ± 2.59),t=-2.813] of students in the case-based learning group were significantly higher than those of the students in the traditional teaching group(P＜0.05).Questionnaire survey showed that the number and proportion of case teaching group of students held a positive attitude towards the teaching methods used in improving students' ability to analyze problems(x2=4.757),increasing study interest (x2=4.371),promoting knowledge mastery (x2=4.276) is higher than traditional teaching group (P＜0.05).Conclusion Case-based learning can inspire students' learning initiative,improve their creative ability to use knowledge,analyze and solve practical problems,so as to promote the course scores.