To study the effect of antioxidants R-(+)-lipoic acid (R-LA) on cells growth,proliferation and related mechanisms in human HepG2 cells lines.MTT was used to measure cells growth and proliferation.Reactive oxygen species (ROS) kit was used to analyze ROS level.Cell apoptosis and cell morphological changes were observed by flow cytometry and Hoechst 33258 test.Protein expression levels of apoptosis,autophagy and related pathway were analyzed through Western blot,including Bax,Bcl-2,caspase 3,PARP,ATG5,ATG7,LC3,Beclin1,mTOR,P70S6K,P38,P53,ERK and Akt etc.Results showed that cell growth and proliferation were inhibited in a dose-and time-dependent manner after being treated by lipoic acid.R-LA could increase ROS production,pro-apoptosis proteins Bax levels,activated caspase family and PARP.Meanwhile,R-LA could up-regulate the levels of autophagy-related proteins including ATG5,ATG7,Beclin1 and LC3,and down-regulate phosphomTOR and P70S6K levels.Signal pathway results showed that R-LA could up-regulate phospho-P38 and phospho-JNK levels,and decrease phospho-Akt and phospho-ERK.When adding 3-methyladenine,autophagy was inhibited.Thus,R-LA might activate autophagy and induce apoptosis by P38/AMPK-JNK,PI3K/AKT and Ras/ Raf/MEK/ERK pathways.

OBJECTIVE: To determine the concentration of phenobarbital(PBB) in serum by RP-HPLC.METHODS: The analytical column was C18.The mobile phase consisted of methanol-water(44∶56) at a flow rate of 1.0 mL?min-1.The detection wavelength was set at 205 nm and the temperature of column was set at 35℃.RESULTS: A good linear relationship was obtained for PBB at a concentration range from 5.08 ?g?mL-1 to 63.50 ?g?mL-1(r=0.999 9).The mean relative recovery was 99.75% and the mean extraction recovery was 96.34%.The intra-day RSD and the inter-day RSD were all less than 4%.CONCLUSION: This method is simple,rapid,accurate,and suitable for therapeutic drug monitoring.

Objective To investigate the effect of folic acid on plasma homocysteine(Hcy) levels and major cardiovascular events in elderly patients with H hypertension.Methods 200 elderly patients with H hypertension were randomly divided into the observation group and control group with 100 cases in each group according to the digital list methods,the control group was given health education and routine anti-hypertensive drug for treatment while the observation group was given folic acid on basis of the conventional treatment,the Hcy levels after 3 months of treatment and incidence of major cardiovascular events among 3 years of the two groups were compared.Results (1) The level of plasma Hcy in the observation group decreased significantly after treatment,with statistically significant difference when compared with before treatment (t =12.32,P ＜ 0.05),that in the control group had no obvious change after treatment than before treatment,there was statistically significant difference between the two groups after treatment(t =11.45,P ＞ 0.05);(2) The occurrence of cardiovascular events among 3 year after treatment of the observation group was 6 cases,the incidence rate was 6％,the control group occurred 15 cases,the incidence rate was 15％,the difference in the incidence of the two groups was statistically significant(x2 =8.24,P ＜ 0.05).Conclusion Folic acid intervention can reduce the plasma Hcy levels in elderly patients with H hypertension and control the occurrence of cardiovascular events.

OBJECTIVETo observe the effects of "menstrual cycle-based acupuncture therapy" on ovarian function and pregnancy results of in vitro fertilization-embryo transfer (IVF-ET) in patients with decline in ovarian reserve (DOR).

METHODSA total of 63 patients of DOR who received treatment of IVF/intracytoplasmic sperm injection (ICSI) were randomly divided into an observation group (30 cases) and a control group (33 cases). The patients in the observation group were treated with "menstrual cycle-based acupuncture therapy". The syndrome differentiation and treatment were given based on different phases of menstruation. Shiqizhui (EX-B 8) and Mingmen (GV 4) were selected during menstrual phase, Shenshu (BL 23), Geshu (BL 17), Sanyinjiao (SP 6) and Taixi (KI 3) were selected after menstruation, Qihai (CV 6), Guanyuan (CV 4), Zigong (EX-CA 1), Zusanli (ST 36) were selected during ovulatory period, Qihai (CV 6), Guanyuan (CV 4), Yanglingquan (GB 34), Taichong (LR 3) were selected before menstruation. The acupuncture was given twice a week until second menstrual cycle of oocyte retrieval. The total times of acupuncture was (15 ± 2). After acupuncture, patients were treated with IVF-ET. The patients in the control group were treated with IVF-ET but no acupuncture. The indices of ovarian reserve function, including basic follicle-stimulating hormone (FSH), estradiol (E2), antral follicle count (AFC), number of retrieved oocytes, number of fertilization and number of high quality embryo, were compared and analyzed before and after acupuncture in the observation group. The differences of outcomes of IVF-ET, including the cycle cancellation rate, implantation rate, the clinical pregnancy rate, were compared between the two groups.

RESULTSCompared before acupuncture, the E2, AFC, number of retrieved oocytes, number of high quality embrgo and number of fertilization were all increased after acupuncture in the observation group (all P< 0. 05). Compared with the control group, levels of the E2, the number of retrieved oocytes, number of fertilization and number of high quality embryo were all increased in the observation group (all P < 0.05). Also, the implantation rate, the clinical pregnancy rate were improved (both P < 0.01) and cycle cancellation rate was reduced (P< 0.01).

CONCLUSIONThe "menstrual cycle-based acupuncture therapy" can effectively improve the ovarian reserve function in DOR patients, leading to an improved clinical pregnancy rate of IVF-ET.

Acupuncture Therapy ; Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; physiopathology ; therapy ; Menstrual Cycle ; Middle Aged ; Ovarian Reserve ; Ovary ; physiopathology ; Pregnancy ; Young Adult

Objeetive To explore the differentiation of B lymphocytes and expression of B7-related protein-1 (B7RP-1)on B lymphocytes in patients with systemic lupus erythematosus(SLE).Methods Three-color immunofluorescent staining and flow cytometric assay were used to analyze the frequency of three types of B lymphocytes,I.e.,plasma cells,memory B lyphocytes and naive B lymphocytes,as well as the expression of B7RP-1 on these cells in peripheral blood from 23 patients with SLE and 16 normal human controls.Clinical data of these patients with SLE were collected.and SLE disease activi index(SLEDAI)was also evaluated.The relationship was assessed between the expression of B7RP-1 and SLEDAI.Results The frequency of plasma cells was highest in patients with active SLE.followed by patients with inactive SLE and normal human controls(P＜0.01).A significant decrease was observed in the frequency of memory B lymphocytes in patients with active SLE compared with normal controls (P＜0.01),but no significant difference was found between patients with inactive SLE and those with active SLE(P＞0.05).Regarding the frequency of naive B lymphocytes,there was no significant difierence among the three groups.Increased frequency of plasma cells was also noted in patients with lupus nephritis(LN)compared with those without LN [(6.15±3.12)%vs(3.31±1.41)%,P＜0.05 ],but no significant difierence was found with regard to the frequency of memory or naive B lymphocytes between these two groups.The expression rate Of B7RP-1 was significantly lower on total lymphocytes from patients with SLE than from normal human controls (46.51%vs 63.75%,P＜0.05),which was the case with B7RP-1 on plasma cells,memory B lyphocytes and naive B lymphocytes (all P＜0.01),whereas no significant difierence was found between patients with inactive SLE and active SLE or between patients with and without LN.In addition.no correlation was found between the expression of B7RP-1 and SLEDAI(r=0.035,P＞0.05).Conclusions In peripheral blood of patients with SLE,the frequency of plasma cells is increased,while the expression of B7RP-1 on lymphocytes is decreased,which may be relevant to the pathogenesis of SLE.

The structure gene pelA from Thermotoga maritima MSB8 encoding pectate lyase was amplified and ligated into pHsh, resulting pHsh-pelA. Through structural optimization on pHsh-pelA, the ultimate plasmid, pHsh-pelC, which possessed the most appropriate structure and free energy of mRNA, was obtained. Pectate lyase C (PelC) was obtained after expressing pHsh-pelC in Escherichia coli JM109. The optimum activity of PelC was determined at pH 8.5 at 90 degrees C, with a half-life for almost 2 h at 95 degrees C. PelC was stable at the pH range of 8.2-9.8, and was dependent on Ca2+ for activity and stability. The enzyme kept stable for a long time and possessed a high level of activity at 60 degrees C. The kinetic assay using polygalacturonic acid (PGA) as substrate gave K(m) and V(max) of 0.11 mmol/L and 327 U per mg of protein. SDS-PAGE analysis showed that the molecular mass of the expressed recombinant PelC was about 43 kD, which was exactly the size predicted. The expression vector system of the heat shock plasmid pHsh owned such advantages as high expression level and cheap induction. Moreover, the superior stability of the recombinant enzyme laid the base for large-scale fermentation application.
Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Hot Temperature ; Polysaccharide-Lyases ; biosynthesis ; genetics ; isolation & purification ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Recombination, Genetic ; Thermotoga maritima ; enzymology

Objective To investigate polymorphism in the promoter region of insulin-like growth factor-1(IGF-1) gene.Methods Five hundred and sixty-one neonates admitted to Peking University Third Hospital from June 1st,2010 to June 30th,2012 were recruited into the study.Gender,gestational age,birth weight and birth length were collected.Heel blood samples were collected on 3-5 days after birth.DNA was extracted to analyze the polymorphism in the promoter region of IGF-1 gene.Chi-square test,independent-sample t-test,analysis of variance and HardyWeinberg equilibrium were performed.Results Among the 561 neonates,413 were full term,and 148 were preterm; 92 were large for gestational age (LGA),433 were appropriate for gestional age (AGA),and 36 were small for gestional age (SGA).Seven different alleles and 23 genotypes in the promoter region of IGF-1 gene were identified in the population.The seven alleles were 188,190,192,194,196,198 and 200 bp respectively.The three most common genotypes were 190-192 bp,192-196 bp and 192-192 bp,whose frequencies were 23.2％ (130/561),15.0％ (84/561) and 12.8％(72/561).There were no significant differences of cytosine-adenosine (CA)19/CA19,CA19/CAno19and CAno19/CAno19 genotypes between full term and preterm infants [11.4％ (47/413) vs 16.9％(25/148),55.9％ (231/413) vs 50.7％ (75/148) and 32.7％ (135/413) vs 32.4％ (48/148)respectively,x2=2.96,1.21 and 0.00,all P＞0.05].There was no difference in the gestional age among infants with CA19/CA19,CA19/CAno19 and CAno19/CAno19 genotypes [(37.1±2.9),(37.6±3.1) and (37.4±3.1) weeks respectively,F=0.54,P=0.58].The frequency of CA19/CA19 in SGA neonates was higher than that in LGA and AGA neonates [25.0％ (9/36) vs 7.6％(7/92) and 12.9％ (56/433),x2 =7.01,P=0.03],but there were no differences in the frequency of CA19/CAno19 and CAno19/CAno19 among LGA,AGA and SGA neonates (CA19/CAno19:x2 =1.13,P=0.57; CAno19/CAno19:x2 =0.58,P=0.75).Conclusions Polymorphism exists in the promoter region of IGF-1 gene.The gestational age is not associated with the frequency of CA19 allele.

Objective The primary culture of synovial fibroblasts is a convenient tool to study the pathology and physiology of synovial tissues .An improved method was constructed in this study by C57BL /6 mice to study the mechanism of rheumatoid ar-thritis(RA) .Methods The synovium around the hip joints were collected .Attention should be paid to eliminate the egg-yolk like yellow oval substance in the middle of the synovium .The synovium was transferred into a 1 .5 mL Eppendorf tube containing 0 .5%type Ⅳ collagenase and cut into 1 mm3 blocks or so .The Eppendorf tube was placed in 37 ℃ Constant temperature orbital shaker incubator for 60 min .After digestion ,the tube was placed on the Vortex for a high-speed oscillation for 1 .5 minutes to guarantee the separation of cells .Results Within about 1 week ,the first passage was performed by the trypsin digestion method .On day 10 , the number of synovial macrophages reached the maximum and then decreased gradually .After the third generation (day 15 to 20) , the synovial macrophages generally disappeared .Vimentin was suitable for the immunofluorescence cytochemical staining for the synovial fibroblasts .The cell purity was indicated as > 95% .The cytometric analysis indicated that purity of Vimentin and CD90 .2-labelled cells was over 95% ;the purity of CD54-labelled cells was 80% approximately .Conclusion It is a simple and effective method for primary culture of synovial fibroblasts in mice .

OBJECTIVE: To compare and analysis outcomes of different age paragraph hearing aid users by questionnaires. METHOD: The questionnaire was deployed to 90 hearing aid users, 40 females and 50 males, with ages between 19 and 80 years-from junior high school education to university education, suffering binaural severe or moderate sensorineural deafness. The subjects were devided into 3 groups, the A group with ages between 19 and 40 years, the B group with ages between 41 and 60 years, the C group with ages between 61 and 80 years. After being fitted with hearing aids for 6 weeks, 6 months and 12 months, SADL and IOI-HA were applied to evaluating 90 subjects. The study was carried out through face to face interview or by telephone. RESULT: There were significant differences in SADL scores of positive effect, service and Personal image among the 3 groups (P < 0.01). The scores of the A group was the highest, and the B group was higher than the C group. On the contrary, there were significant differences in SADL scores of negative effect among the 3 groups (P < 0.01), with the scores of the A group being the lowest, and the B group being lower than the C group. There were also significant differences in IOI-HA scores among the 3 groups (P < 0.01). The scores of the A group was the highest, and the B group was higher than the C group. CONCLUSION The outcome and satisfaction of hearing aids among different age paragraph is improved with the wearing time growth, but the outcome and satisfaction of hearing aids are decreased with the age growth.
Adult ; Aged ; Aged, 80 and over ; Female ; Hearing Aids ; Hearing Loss, Sensorineural ; rehabilitation ; Humans ; Male ; Middle Aged ; Patient Satisfaction ; Self Concept ; Surveys and Questionnaires ; Young Adult

Objective: To evaluate the clinical effect of Jinghua Weikang capsule in treating peptic ulcer with intermingled heat and cold syndrome. Methods: From January 2016 to December 2018, 90 patients of peptic ulcer with intermingled heat and cold syndrome in Changzhou Hospital of Traditional Chinese Medicine were divided into two groups by the random digital table method, with 45 cases in each group.The control group was treated with rabeprazole capsule, and the observation group were treated with rabeprazole capsule and Jinghua Weikang capsule.The gastroscopic performance, the main individual symptoms and TCM syndrome between the two groups were compared. Results: The gastroscopic performance[42 cases(93.3%) were clinical cured, 1 case(2.2%) was significantly improved, 1 case(2.2%) was improved], TCM syndrome effect[38 cases(84.4%) were clinical cured, 4 cases(8.9%) were significantly improved, 2 cases(4.4%) were improved] in the observation group were significantly better than those in the control group[the gastroscopic performance: 35 cases(77.8%) were clinical cured, 5 cases(11.1%) were significantly improved, 3 cases(6.7%) were improved], TCM syndrome effect: 28 cases(62.2%) were clinical cured, 6 cases(13.3%) were significantly improved, 8 cases(17.8%) were improved](Z=-2.040, -2.463, all P<0.05). The improvement of TCM symptoms of epigastric pain which could be relieved by warmth and pressure, dry mouth with bitter taste, cool limbs were significantly better than those in the control group(χ²=6.154, 6.056, 7.174, all P<0.05). Conclusion Jinghua Weikang capsule can accelerate the repair of gastric mucous of patients with intermingled heat and cold syndrome, improve clinical syndrome, especially these symptoms: stomacha which can be relieved by warmth and pressure, dry mouth with bitter taste, cool limbs.