Objective: To construct mcpr1prokaryoti c expression vector and to express MCPR1 protein.Methods:PCR was used to obtain coding region of mcpr1. Construction of a high-level fusion protein expression vector pGEX-4T-mcpr1 was conducted by inserting the fra gment of coding region of mcpr1into a fusion protein expression vector pGEX -4T-1. Then the recombinant plasmid was transferred into E. colito prepar e the MCPR1/GST fusion protein. DNA sequencing and endonucleases digesting were used to check the coding region. Results:pGEX-4T- mcpr1 wa s constructed successfully and the coding region was inserted into the vector co rrectly. A new protein band of 36 000 was observed by SDS-PAGE analysis after i nduction by IPTG. The 36 000 protein amounted to 39 percent of the total prote in and existed mostly in precipitation of broken bacteria. Conclusion: MCPR1 protein can be expressed in E. coliexpression system and purif ied initially.

In this paper, the specific male sequeuce in paraffin section of various tissues with autolys is ofdifferent degrees was detected by polymerase chain reaction(PCR).The results shewed that PCR could be used for identifying sex in the tissues with low degrees ofautolysis; its positive rate was lower in high degrees with disappearance of nuclaic member and cell ou-tline,and often gave false negative results when the autolysis degree was high.

Objective To systematically evaluate the safety of high dose atorvastatin (80 mg daily) in Chinese patients. Methods Randomized controlled trials (RCTs) investigating 80 mg/ d atorvastatin vs. low-dose atorvastatin or placebo or blank were electionically retrieved in date bases of EMbase, PubMed, the Cochrane Library, WanFang, CNKI and WeiPu. Meta-analysis was performed using RevMan 5. 2 and Stata 11. 0 software. Results A total of 20 RCTs involving 2282 cases were included. The results of meta-analysis showed no significant differences betweent the 80 mg/ d atorvastatin group and the control group in the incidence of gastrointestinal adverse events (RR 1. 53, 95% CI 0. 85-2. 76, P = 0. 16), hepatic adverse events (RR 1. 53, 95% CI 0. 99 - 2. 36, P = 0. 05), muscular adverse events (RR 1. 51, 95% CI 0. 92 -2. 49, P = 0. 10), serious hepatic injuries ( RR 2. 33,95% CI 0. 88 - 6. 20, P = 0. 09) and serious muscular myopathies (RR 1. 40, 95% CI 0. 46 - 4. 30, P = 0. 56). Subgroup analysis by type of cotrast media used and durations of taking 80 mg/ d atorvastatin showed there were higher risks of gastrointestinal adverse events in the 80 mg/ d group when compared to blank control ( RR 4. 22, 95% CI 1. 11 - 16. 04, P = 0. 03). Conclusions The current evidence shows that 80 mg / d atorvastatin may be relatively safe in terms of adverse events in gastrointestinal tract, liver and muscular system, and relatively has risk in causing severe liver injuries and myopathies. With limited quantity and quality from the RCTs available, more high quality RCTs are needed to verify the above conclusion.

Objective To explore the early diagnosis and treatment of congenital mesenteric hiatual hernia.Methods A retrospective study was carried out in 4 patients with congenital mesenteric hiatual hernia in Tongji hospital from Nov.2005 to Mar.2007,and combining lite-rature,the diagnosis and treatment of mesenteric hiatal hernia was summed up.Results Four patients were diagnosed in operation.One case was thought as adhesive intestinal obstruction before operation;two patients were on emergency operation and 2 patients were on time-elective operation;one patient preoperative CT scan may suggest mesenteric hiatal hernia;one case had partial resection of small intestinal,the others were replaced the intestine and fixed the defect.One patient occurred early septic shock;all of them had get well.Conclusions It′s hard to diagnose the congenital mesenteric hiatual hernia before operation.Abdomen CT examination and multislice spiral CT angiography (MSCTA) help to diagnose.Early diagnosis and timely operation are the therapeutic key of congenital mesenteric hiatual heria.For the patients with recurrent abdominal pain,who was not confirmed with a variety of inspection,laparoscopic exploration can provide diagnosis,and can take the initiative to control the development of disease.

Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts ; Gene Expression Regulation, Plant ; Glycyrrhiza uralensis ; genetics ; Plant Roots ; Sequence Analysis, DNA ; Stress, Physiological ; Transcriptome

AIM: To explore the effect of melatonin (MLT) on the apoptosis of thymocytes and splenocytes in mice induced by ionizing radiation and its mechanism. METHODS: The percentages of apoptotic bodies and the DNA lytic rates of thymocytes and splenocytes in mice in vitro and in vivo were detected with flow cytometry and fluorospectrophotometry, respectively. RESULTS: The apoptosis of mouse thymocytes and splenocytes in vitro increased with significant dose-dependence in 0 5-6 0 Gy X-irradiation. When MLT of 2 mmol?L -1 was added into thymocytes or splenocytes in vitro before irradiation with 0 5-6 0 Gy X-rays, the percentages of apoptotic bodies and the DNA lytic rates all decreased significantly as compared with those in the irradiation group. The percentages of apoptotic bodies in these two kinds of cells were 86 25% and 89 22% of those in the irradiation group, respectively, and the DNA lytic rates were 87 23% and 89 16%, respectively. When MLT was injected into intraperitonium in mice 60 min before whole-body irradiation with 2 Gy X-rays, the percentages of apoptotic bodies and the DNA lytic rates were significantly lower than those in the irradiation group, and near or lower than those in the sham-irradiation group. MLT of 0 1-2 5 mg/kg decreased the lymphocyte apoptosis, but without significant dose-dependence. CONCLUSION: The protective effects of MLT on mouse lymphocytes damaged by irradiation in vivo are obvious than those in vitro. [

Objective: To investigate the value of the trigeminocervical response (TCR) for revealing bulbar involvement in patients with spinal and bulbar muscular atrophy (SBMA). Methods: Thirty patients with SBMA and 30 healthy male controls were included in this study. In all of the normal controls, stimulation of the infraorbital nerve on one side produced bilateral short latency waves consisting of a positive/negative wave, p19/n31, the mean latency of which was measured. The mean square root of the ratio between the amplitude of p19/n31 and the mean rectifi ed surface electromyography (EMG) activity preceding the stimulus, the A value, was estimated. The parameters of the TCR were compared between the two groups. Results: Among the patients with SBMA, 21 (70.0%) had delayed latencies of p19/n31 (P < 0.01) and all (100%) had reduced A values (P < 0.01) relative to the normal controls. Conclusions: All parameters of the TCR were signifi cantly different between the patients with SBMA and the normal controls. T

OBJECTIVE:To investigate improvement effect of Duzhong zhuanggu(DZ)pills on retinoic acid-induced osteopo-rosis in mice. METHODS:72 KM mice were randomized into blank control group(normal saline),model group(normal saline), positive control group (Xianling gubao capsule,0.50 g/kg),DZ pills high-dose,medium-dose and low-dose groups (1.90,0.95,0.48 g/kg,DZ-H,DZ-M,DZ-L groups),with 12 mice in each group. Except for blank control group,other groups were given ret-inoic acid intragastrically to induce osteoporosis model,and given relevant medicine intragastrically once a day,for 14 days. After last administration,body weight of mice was determined;the blood sample was collected from eyeball to determine the contents of serum calcium and phosphor;liver,spleen and thymus gland were collected to determine organ index;left femur was collected to determine bone wet weight,bone dry weight,length and diameter;right femur was collected to determine the area percentage, width and number of bone trabecula. RESULTS:Compared with blank control group,the body weight of mice in model group were decreased;the contents of serum calcium and phosphorus were increased;liver indexes and spleen indexes were increased, while thymus indexes were decreased;bone wet weight,bone dry weight,length and diameter were decreased;area percentage, width and number of trabecular were decreased,with statistical significance (P<0.05 or P<0.01). Compared with model group, the contents of serum calcium were decreased in treatment groups,and the contents of serum phosphorus were decreased in positive control group,DZ-M and DZ-L groups. Thymus indexes were increased in positive control group,DZ-M and DZ-L groups,while liver indexes were decreased in positive control group;femur wet weight of positive control group as well as femur dry and wet weight,diameters of DZ-M were all increased. The percentage,width and number of bone trabecula were increased in positive con-trol group,DZ-H and DZ-M group,and the number of bone trabecula was increased in DZ-L group,with statistical significance (P<0.05 or P<0.01). CONCLUSIONS:DZ pills have improvement effect on osteoporosis induced by retinoic acid in mice.

Objective To observe the curative effect of autologous bone marrow stem cells implantation to bone inducing active material combined with core decompression in the treatment of early femoral head osteonecrosis (FHON).Methods From April,2010 to March,2012,in Department of Orthopaedics,the First Affiliated Hospital of Zhengzhou University,a total of 79 adult patients with 108 hips suffered from the early stage FHON were treated with autologous bone marrow stem cells implantation to bone inducing active material combined with core decompression through the core of the femoral canal,male of 65 cases,female of 14 cases,the mean age was 29.5 (20-50) years old.According to the etiology classification:the alcohol-induced FHON was in 54 patients with 66 hips,steroid-induced FHON in 14 patients with 20 hips,steroid and alcohol-induced ONFH was in 11patients with 22 hips.According to association research circulation osseous (ARCO)classifying,Ⅰ-A,Ⅰ-B,Ⅱ-A,Ⅱ-B phases were 6,16,8,and 78 hips,respectively.There were 43 hips in left side and 65 hips in the right side.Results All patients were followed up from 4 to 6 (4.8 ± 0.6) years.Compared with before operation,the scores of all patients were significantly increased (P ＜ 0.05).All patients with hip pain symptoms were relieved or disappeared.The healing tine of the patients in all age groups was statistically significant (P ＜ 0.05),and with the increase of age,the healing time was prolonged.The excellent and good rates of Ⅰ-A,Ⅰ-B,Ⅱ-Aand Ⅱ-B were 100％ (6 / 6),100％ (16/16),100％ (8/8),and 98.7％ (77/78).The X-ray showed that coarse channel osteogenic phenomenon is obvious,there is 1 case collapse of femoral head of stage Ⅱ-B,the rest were not collapse.Conclusion The treatment of early osteonecrosis of the femoral head with autologous bone marrow stem cells implantation to bone inducing active material combined with core decompressionis definitely effective,especially in patients with ARCO:Ⅰ-A,Ⅰ-B and Ⅱ-A phase,and the effect of ARCO:Ⅰ-A and Ⅱ-A is the best.

AIM:To construct adenovirus vector expressing mice B7-H1 gene, transfect dendritic cells ( DCs ) , and to study the therapeutic effect of modified DC on thyroid-associated ophthalmopathy ( TAO) in mice.
METHODS: We designed and constructed B7-H1 gene adenovirus expression vector, and transfected DCs from mouse bone marrow, tested the phenotype and function of modified DCs, identificated its negative regulation to immune responses. The modified DCs were infected the sicked mice. And then the immunotherapeutic effect of modified DCs to TAO were tested.
RESULTS: B7 - H1 gene adenovirus vector was constructed and transfected DCs from bone marrow. The titer of the recombinant adenovirus was 1. 8í109 PFU/mL. B7-H1 gene modified DCs characteristics of regulatory DCs, could inhibit positive immune responses. The inhibition proceeding of TAO into mice infected modified DCs, was obviously prior to the control mice. The gene modified DCs, maybe become the new immunotherapy biological agent to thy TAO.
CONCLUSION: We constructed the expression of mouse B7 - H1 gene adenovirus expressed vector successfully, transfected DCs, by vector have properties of regulatory DCs, inhibiting positive immune response and the occurrence and development of thyroid eye disease. Gene modified DCs, reveal potent to the treatment of thyroid eye disease.