In order to improve the expression level of segment of GABAA receptor a1 subunit in E. coli, growth conditions of the recombatant, which influence the final yield of protein expression, including growth medium, inoculation ratio, temperature, pH, rotation speed, inducing time and concentration of IPTG and so on, were studied in shaking flasks. The results indicated that, with 3% inoculation ratio, cultured 3.5 hours at 37℃, and then induced 5 hours by IPTG at 32℃, the yield of GABAA receptor protein was 95mg/L and the biomass was 3.25 g/ L. In contrast, using a 16 L stirred fermentor instead of shaking flasks, the highest level of the protein expression, 136mg/L with 4.95g/L of biomass, was achieved after fermenting 5.5 hours.

Objective To explore the psychomotor development in 12 month old infants born after in vitro fertilization with a control group of infants concieved naturally.Methods A matched control study was performed on psychomotor development of 12-month-old infants concieved with use of assisted reproductive technology(in vitro fertilization only).The control group was matched according to maternal age,parity,social class and level of parental education.The suitable mothers were invited to participate at the 28th week of gestation and were followed up to delivery.The infants of the two groups were followed up to 12 months old and a formal developmental assessment was done with the Children Development Center of China(CDCC) scales of infant development.Results The incidences of preterm birth,low birth weight and twin pregnancy were significantly higher in the assisted conception group.No statistically significant difference was found in the mental development index and psychomotor development index between assisted conception and control groups.Conclusions The levels of psychomotor development in 12 months old infants born after in vitro fertilization are normal.But as the incidences of preterm birth,low birth weight and multifetation are significantly higher in vitro fertilization group,it need to follow up the ongoing development of these children.

4-Hydroxycoumarins ; blood ; Animals ; Chromatography, High Pressure Liquid ; methods

Animals ; Carps ; physiology ; Galvanic Skin Response ; drug effects ; physiology ; Skin ; cytology ; Strychnine ; pharmacology ; Synaptic Transmission ; drug effects ; physiology

Objective To evaluate the value of 18F-FDG PET/CT in assessing radiosensitivity enhancement by irisquinone (IR) on rabbit xenografted VX2 lung tumor models.Methods Twenty-four tumor-beating rabbits were randomly divided into 3 groups (8 rabbits/group):group A with radiotherapy alone,group B with combined radiotherapy and IR,and group C without radiotherapy (the control group).18F-FDG PET/CT imaging was performed before radiotherapy and 24 h and one week after radiotherapy.The tumor SUVmax on delayed imaging was calculated in all rabbits.Two rabbits in each group were sacrificed after PET/CT imaging.HE staining was used to assess the differences in cancer cells among groups.Paired t test,one-way analysis of variance and Kaplan-Meier analysis were performed to analyze the data using SPSS 13.0.Results Before radiotherapy,the tumor SUVmax of all the 24 rabbits on standard and delayed imaging were 2.200 ± 0.761 and 3.162 ± 0.833 (t =-5.582,P ＜ 0.01).At 24 h post-radiotherapy,the delayed SUVmax of groups A,B and C were 2.614 ± 0.654,2.349 ± 0.869 and 5.663 ± 1.144,respectively.The differences between pre-radiotherapy and 24 h post-radiotherapy were statistically significant in all three groups (t =2.527,3.620,11.011,all P ＜0.05).One week after radiotherapy,the delayed SUVmax of groups A,B and C were 3.625 ± 1.064,3.058 ±0.850 and 7.424 ± 1.751,respectively.The differences among groups A,B and C were statistically significant (tA∶ B =2.652,tA∶C =3.799,tB∶C =4.366,all P ＜0.05).The cancer cells of group B were fewer than those of groups A and C by pathological findings,which was consistent with 18F-FDG PET/CT results.The survival times of groups A,B and C were (62.375 ±4.534),(69.000 ±4.660) and (54.125 ±5.276) d,respectively.Kaplan-Meier survival curves revealed better survival of group B as compared to groups A and C,respectively (Log-rank,x2 =7.355,16.943,both P ＜ 0.01).Conclusion 18 F-FDG PET/CT is able to evaluate the effect of irisquinone on tumor radiosensitivity enhancement.

Objective To explore the apoptosis of alveolar epithelial cell(AEC)and the dynamic changes of Caspase-3 mRNA and Bax and Bcl-2 expression in premature rat with hyperoxia-induced chronic lung disease(CLD).Methods Sixty premature rats within 1 day after birth were randomly divided into 2 groups:hyperoxia group(n=30)and control group(n=30).Hyperoxia-induced premature rat CLD models were prepared,and situ nick end-labeling(TUNEL),reverse transcription polymerasechain reaction(RT-PCR)and immunohistochemical technique were used to determine apoptosis index(AI)of AEC and the expressions of Caspase-3 mRNA and Bax and Bcl-2 proteins in lung tissues at 1,3,7,14 and 21 d after birth.Results Compared with control group,in the hyperoxia group,on the third day after exposured to hyperoxia,the AI of AEC and the expressions of Caspase-3 mRNA and Bax began to increase,and the expression of Caspase-3 mRNA was kept at high level on 7-21 d.The expression of Bcl-2 began to decrease on 7 d,and significantly decreased on 7-21 d.AI of AEC was positively correlated with the expression of Caspase-3 mRNA and Bax,and negatively with the expression of Bcl-2.Conclusions Hyperoxia may induce the increased expression of Caspase-3 mRNA,which might result in the abnormal expression of Bax and Bcl-2 in lung tissues and their imbalance,which might be one of the underlying mechanisms of apoptosis of AEC in premature rats with CLD.

AIM: To compare the effectiveness of botulinus toxin of type A and complete resection of the periorbital muscle on idiopathic blepharospasm.
● METHODS: Patients with idiopathic blepharospasm and having undergone either of two procedures from Dec. 2010 to Jun. 2015 were selected ( 60 patients ) . Among them, group A (30 patients, 60 eyes) underwent botulinus toxin of type A, group B (30 patients, 60 eyes) underwent complete resection of the periorbital muscle.
●RESULTS: ln group A, the patients with complete response, obvious response, partial response, and no response were 36(60. 0%), 20(33. 3%), 2(3. 3%) and 2 (3. 3%) cases respectively. ln group B, the patients with complete response, obvious response, partial response, and no response were 16(26. 7%), 24(40. 0), 12(20. 0%) and 8 ( 13. 3%) cases respectively. The difference was statistically significant ( Z = - 2. 968, P = 0. 003 ). The relapse rate of group A and group B were 93. 3% and 20. 0% after 6mo, the difference was statistically significant (χ2=32. 851, P<0. 001).
●CONCLUSION: The botulinus toxin injection of type A is effective for idiopathic blepharospasm. But recurrence rate is high after 6mo. Complete resection of the periorbital muscle have long-term efficacy for idiopathic blepharospasm. It′s a supplementary therapy after idiopathic blepharospasm recurrence.

Objective To analyze the differential proteomics of ASMC stimulated by wild IL-13 and mutant IL-13 and to investigate the relations of protein profiles of ASMC to asthma and possible targets for the treatment of bronchial asthma.Methods The total proteins of ASMC stimulated by wild IL-13 and mutant IL-13 were separated by immobilized pH gradient(IPG)-based 2-DE and the differentially expressed protein spots were identified by matrix assisted laser desorption-time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE detected approximately(840?21)spots on wild IL-13 samples and(892?17)spots on mutant IL-13 samples(n=3)and(685?19)spots matched.Six significantly differential proteins were subjected to MALDI-TOF-MS analysis and three of them were identified as stathmin 1,Ribosomal protein p~0 and NADH dehydrogenase.Conclusions ASMCs stimulated by wild IL-13 and mutant IL-13 present different proteomic profiles that may shed some light on the mechanism for the asthma causing effect of wild IL-13 and mutant IL-13.

OBJECTIVETo study the effects of Citalopram on the mRNA expression of bax and bel-2 in frontal cortical neurons and on cell apoptosis of rats after stress.

METHODSTwenty-four healthy male SD rats were randomly divided into three groups (n = 8). The control group did no receive any treatment, the stress group was subject to stress and given normal saline and experimental group was given Citalopram irrigation stomach after stress. Rats were forced to swim to establish chronic stress model (15 min/d, 4 weeks), bax, bcl-2 mRNA expression were tested by in situ hybridization technique (ISH), TUNEL assay was used to determine cell apoptosis, Nikon image analysis software were used to measure the number of positive cells in each index.

RESULTSCompared with the control group, the stress group showed a larger number of bax mRNA expressing cells( P < 0.01), a smaller number of bcl-2 mRNA expressing cells (P < 0.01), and the staining intensity of positive cells was significantly reduced( P < 0.01). Compared with the stress group, the experiment group showed more reduced number of bax mRNA positive cells( P < 0.01) and significantly increased bcl-2 mRNA positive cells( P < 0.05), a small amount of positive cells were found, compared with that in the stress group, nuclear condensation in the experimental group was reduced significantly and the staining was obviously weaker( P < 0.01).

CONCLUSIONCitalopram significantly antagonizes bax mRNA and potentiatesbcl-2 mRNA protein expression and inhibits apoptosis of rat prefrontal cortical neurons caused by chronic stress, which might be one possible mechanism of Citalopram for prevention and treatment of psychosis caused by chronic stress.

Animals ; Apoptosis ; Citalopram ; pharmacology ; Male ; Neurons ; drug effects ; metabolism ; Prefrontal Cortex ; cytology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Stress, Physiological ; bcl-2-Associated X Protein ; metabolism

Objective: To observe the difference of the effect of low-frequency electroacupuncture (EA) on blood lipids between male and female patients with simple obesity due to damp induced by spleen deficiency. Methods: Eighty patients with simple obesity were recruited, including 37 males and 43 females, to receive low-frequency EA by selecting Yinlingquan (SP 10), Sanyinjiao (SP 6), Zusanli (ST 36), Fenglong (ST 40), Quchi (LI 11), Tianshu (ST 25), Zhongwan (CV 12), Shuifen (CV 9), Qihai (CV 6) and Guanyuan (CV 4), with needles retained for 30 min. The treatment was given once a day, 10 sessions as a treatment course, for 2 courses in total. The contents of body fat percentage (F%), total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), blood glucose (Glu) and adiponectin (ADPN) in serum were observed to see the changes, and the two groups were compared and analyzed. Results: After the treatment, F%, and serum contents of TC, TG, LDL, Glu and ADPN dropped significantly in the two groups (P<0.05 or P<0.01) and the serum content of HDL increased significantly in male group (P<0.05). The decrease of F% in female group was more significant than that in male group (P<0.01); the decrease of ADPN in male group was more significant than that in female group (P<0.05). Conclusion: EA can regulate the disordered blood lipids in male and female patients with simple obesity, with certain differences between genders. The decrease of subcutaneous fat content is more significant in females than that in males, while the decrease of ADPN is more significant in males.