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.

Adult ; Diagnosis, Differential ; Endometriosis ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; pathology ; Lumbar Vertebrae ; pathology ; Magnetic Resonance Imaging ; Spinal Canal ; pathology ; surgery ; Spinal Diseases ; pathology ; surgery

Objective To investigate the effect of acupuncture at Zusanli, Sanyinjiao and Danzhong on mammary estrogen receptor (ER) and progesterone receptor (PR) expressions in rats with dimethylbenzanthracine (DMBA)-induced mammary cancer. Methods One hundred and twenty female SD rats aged 6-8 weeks were randomized into a model group of 60 rats and a blank group of 30 rats. The model group received an oral gavage of DMBA for model making. The blank group received an oral gavage of equal volume of sesame oil. At 15 weeks after model making, the model group of rats was randomized into treatment and control groups. The treatment group received acupuncture at Zusanli, Sanyinjiao and Danzhong, and the control and blank groups, only the same grasp and release. After the completion of acupuncture treatment (twenty-seventh week), abdominal venous blood was taken and serum tumor markers were determined by electrochemiluminescence immunoassay. Tumor masses were counted and their shapes were recorded. The mass was taken and its height, maximum diameter and vertical diameter were measured using a 1 mm precision vernier caliper. Pathological changes in tumor tissues, and ER and PR positive areas and mean optical densities were observed under an Olympus optical microscope.Results There were statistically significant post-treatment differences in the average number and volume of mammary tumors between the treatment group and the control or blank group (P<0.01,P<0.05) and between the control and blank groups (P<0.01). There were statistically significant post-treatment differences in the concentrations of various tumor markers (CA724, CA125, CA199, AEP, CA15-3, CEA and CA50) between the treatment or control group and the blank group (P<0.01,P<0.05) and between the control and blank groups (P<0.01). There was a statistically significant post-treatment difference in CA15-3 concentration between the treatment and control groups (P<0.01). There were statistically significant post-treatment differences in ER and PR positive areas and mean optical densities between the treatment group and the control or blank group (P<0.01) and between the control and blank groups (P<0.01).Conclusions Acupuncture can reduce the occurrence of rat DMBA-induced mammary tumor (including the number and volumes of the tumors). The mechanism of its action may be related to decreasing the concentrations of tumor markers CA724, CA125, CA199, CA15-3, AEP, CEA and CA50 and especially to decreasing CA15-3 concentration, and ER and PR positive areas and mean optical densities.

Objective:To observe the therapeutic effect of electroacupuncture (EA) at Zusanli (ST 36),Guanyuan (CV 4) and Ashi points on adjuvant arthritis rats,and explore the mechanism of EA treatment of rheumatoid arthritis (RA).Methods:Sixty male rats were randomly divided into a normal group,a model group,a methotrexate group and an EA group,with 15 rats in each group.Rats in the normal group and the model group were routinely raised and did not receive treatment;rats in the methotrexate group received methotrexate at a dose of 0.35 mg/(kg·bw),twice a week for 3 weeks;rats in the EA group received acupuncture at Zusanli (ST 36),Guanyuan (CV 4) and Ashi points,and the bilateral Zusanli (ST 36) and Ashi points were connected to EA apparatus,once a day for 3 weeks.The general status,the swelling degree of the toe,the arthritis index (AI) score,the pathological morphology of the ankle joint,and the mRNA expressions of cellular inhibitor of apoptosis protein (c-IAP) 1 and c-IAP2 in joint synovial tissue cells of the rats in each group were observed.Results:The swelling degree of the toe,AI score and mRNA expressions of c-IAP1 and c-IAP2 in the model group were significantly higher than those in the normal group (all P＜0.05).Compared with the model group,the swelling degree of the toe,AI score and mRNA expressions of c-IAP1 and c-IAP2 in the methotrexate group and the EA group improved (P＜0.01 or P＜0.05);the expressions of c-IAP1 mRNA and c-IAP2 mRNA in rat synovial tissues in the EA group were significantly higher than those in the methotrexate group (P＜0.01).Conclusion:EA alleviates joint swelling in rats with adjuvant arthritis.The mechanism may be related to suppressing mRNA expressions of c-IAP1 and c-IAP2,thus to induce apoptosis of synoviocytes.

Background The molecular biological mechanisms of diabetic retinopathy(DR)is unclear up to now.Researches have proved that endoplasmic reticulum stress(ER Stress)-associated factors are elevated in peripheral blood in patients with diabetic retinopathy.and P58 IPK can inhibit those factors.So the relationship between P58 IPK and DR is worth to investigate. Objective The aim of this study was to detect the dynamic expression of P58 IPK in the retina of diabetic rats. Methods The diabetic animal models were established in 18 clean male SD rats by intraperitoneal injection of stilptozotiein(STZ)at a dose of 60 mg/kg.The rats were sacrificed in 1,3,6 months after injection.The expression change of P58 IPK mRNA in the rats retina was detected by quantitative real-time RT-PCR.Other 6 matched normal rats were used as control groups.This experiment followed the Regulations for the Administration of Affair Concerning experimental Animals by State Science and Technology Commission. Results The rats showed more drinking,more food and more urine after STZ injection with the blood glucose level≥ 16.5 mmol/L.The success rate of diabetic models was 100%.The A value of P58 IPK mRNA/β-actin in rat retina was 0.800±0.005 and 0.975±0.008 after injection of STZ.and that of control rats was 0.725±0.006,showing statistically significant difference between them(t=22.589,t=62.784,P＜0.05).In 6 months after injection of STZ,the expression of P58 IPK mRNA in experimental diabetic rat retina was evidently lower than the eontrol rats(0.671±0.004 versus 0.725±0.006,t=-17.984,P＜0.05).Conclusion P58 IPK has a close relation to the pathogenesis of DR,and it plays a retarding role for DR.

Objective To introduce the clinical experience and investigate venous drainage of distal- ly-based dorsal digital neurocutaneous flap for finger-pulp reconstruction.Methods From Mar.2004 to Oct.2005,18 patients with traumatic finger-pulp defect (＞2cm) were treated by distally based dorsal ho- modigital neurocutaneous flaps.The flap measured 2 cm?2cm～3cm?4cm with the neuro-veno-adipal ped- icle 1cm wide and 2～3cm long.The pivot points were proximal to the PIP joints.The dorsal digital nerve was neurorrhaphied with the proper digital nerve of the recipient site.The dorsal digital vein was ligated at 1cm distal to the pivot point to prevent venous ingress.No venous anastomosis was performed.Results All the flaps survived but had some degrees of venous congestion and swelling,and 8 flaps developed tension blisters. In 13 flaps with follow-up more than 6 months,protective sensation was restored.Conclusion Dorsal digit- al neurocutaneous flap is simple and effective for finger-pulp reconstruction.Ligating the big superficial vein at the distal base to interrupt venous ingress,and allowing the proximal vein open and venous egress,can reduce flap congestion and swelling.

Objective To construct the molecular transmission networks of HIV-1 CRF01 AE strains circulating in China and to analyze their characteristics. Methods Sequences of the pol genes of Chinese CRF01 AE strains were downloaded from Los Alamos database and the related demographic informa-tion was checked. Transmission networks were created by four steps,including phylogenetic tree construc-tion,transmission cluster extraction,minimum genetic distance identification and network visualization. Chi-square test was performed to analyze the differences in the distribution of different populations in the networks and the differences in the distribution of subjects with different degrees among different sub-populations. De-scriptive analysis was used to investigate the transmission links between sub-populations and various regions. Results The 2 419 sequences constituted 250 separate networks including 847 nodes and 610 edges. The number of subjects covered by different networks ranged from 2 to 25. Subjects with degree ≥2 represented only 26. 4%(224 / 847)of network-individuals,but were connected with 66. 5%(563 / 847)of all network-individuals. There were 37. 6%(669 / 1 781)of men who have sex with men(MSM),28. 4%(102 / 359) of heterosexual men,16. 1%(10 / 62)of intravenous drug users(IDUs)and 30. 4%(66 / 217)of hetero-sexual women involved in the networks(χ2 =23. 774,P﹤0. 001). The percentage of subjects with degree ≥2 was high in MSM(10. 8% ,193 / 1 781)compared to that in heterosexual men(5. 6% ,20 / 359)(P =0. 002)and heterosexual women(4. 6% ,10 / 217)(P=0. 004). Of the 669 MSM in the networks,95. 5%(639 / 669)linked to other MSM and only 2. 4% (16 / 669)linked to heterosexual women. However, 15. 1%(10 / 66)of the heterosexual women in the networks linked to MSM. Of the heterosexual men in the networks,35. 3%(36 / 102)linked to heterosexual women,9. 8%(10 / 102)linked to MSM. Of the sub-jects in the networks,20. 9%(177 / 847)linked to other regions' individuals. Conclusion The super-spreaders played an important role in the molecular transmission networks of HIV-1 CRF01 AE strains even though they were in a minority. The transmission of HIV-1 CRF01 AE strains between sub-populations and various regions was complicated and active.

OBJECTIVETo investigate genotype of HCV infection among residents in a rural village, Hebei Province.

METHODSBlood samples of the 520 residents were collected, and C/E1 of HCV genome of the 483 samples were obtained by RT-PCR amplifying, and the gene sequences were analyzed and the polygenetic tree were drawn by the software Mega 4.0.

RESULTSIn 483 subjects, HCV-RNA positive objectives are 70, positive prevalence is 14.5%, genotyping for all the samples successfully, including genotypes 1b and 2a, which are 36 (51.4%), 34 (48.6%) respectively.

CONCLUSIONHCV RNA positive rate is 14.5%, which is higher than general people. Subtypes 1b and 2a seemed to be the dominant genotypes of HCV in Zhao county area of Hebei Province.

Adolescent ; Adult ; Blood Donors ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; isolation & purification ; Hepatitis C ; epidemiology ; virology ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Molecular Sequence Data ; Phylogeny ; Young Adult

OBJECTIVEAttention deficit hyperactivity disorder (ADHD) is one of the most common behavior disorders in childhood and adolescent. The etiology of ADHD is unknown. The aim of this study was to investigate the relationship between each of the 14 polymorphisms in the five candidate genes and ADHD, and between the combination of some polymorphisms in those genes and ADHD, in attempting to examine whether combinations of genotypes would confer a significant susceptibility to ADHD.

METHODSOne hundred and thirty-nine children with ADHD and one hundred and nineteen normal children were enrolled. Eight single nucleotide polymorphisms (SNP) of three candidate genes were examined with PCR and RFLP techniques. 48 bp VNTR in DRD4 gene was examined with PCR, nondenaturing polyacrylamide gel electrophoresis and silver staining. Five microsatellites (MS) of three candidate genes were examined with genotyping. The relationship between the combinations of 12 polymorphisms and ADHD was examined with logistic regression analysis.

RESULTS1.The frequency of 1065T/1065T genotype and the 1065T allele were significantly higher in ADHD children than that in normal controls (P<0.05). The frequency of -48G/-48G genotype of the A-48G polymorphism of DRD1 gene was significantly lower in ADHD children than that in normal controls (P<0.05). 2. A specific combination of three polymorphisms in the two genes showing an association with ADHD gave a prediction level of 77.5%.

CONCLUSIONSThe T1065G polymorphism in the SNAP-25 may be associated with ADHD. The 1065T/1065T genotype and the 1065T allele may be a risk factor for ADHD. The A-48G polymorphism of DRDI may be associated with ADHD. The -48G/-48G genotype may be a protective factor for ADHD. The specific combination of three sites of SNP in SNAP-25 gene and DRDI gene is found and shows an association with ADHD in 12 polymorphisms of the five candidate genes on glutamatergic/dopaminergic pathway.

Adolescent ; Attention Deficit Disorder with Hyperactivity ; genetics ; Child ; Female ; Humans ; Logistic Models ; Male ; Minisatellite Repeats ; Polymorphism, Single Nucleotide ; Receptors, Dopamine D3 ; genetics ; Receptors, Dopamine D4 ; genetics ; Receptors, Dopamine D5 ; genetics ; Receptors, N-Methyl-D-Aspartate ; genetics ; Synaptosomal-Associated Protein 25 ; genetics

OBJECTIVETo investigate the changes in the levels of interleukin-17 (IL-17) and transforming growth factor beta 1 (TGF-β1) in serum and bronchoalveolar lavage fluid (BALF) and their clinical significance among children with asthma.

METHODSFifty-six children with asthma were divided into moderate or severe asthma (n=37) and mild asthma groups (n=19) and 18 children without asthma were selected as the control group. Cells in BALF were counted under a microscope. The levels of IL-17 and TGF-β1 in serum and BALF were measured using ELISA.

RESULTSwere no significant differences in total cell count and percentage of macrophages between the two asthma groups and the control group (P>0.05). The percentages of neutrophils, eosinophils and epithelial cells in BALF were significantly higher in the two asthma groups than in the control group (P<0.05). The two asthma groups had significantly higher levels of IL-17 and TGF-β1 in serum and BALF than the control group (P<0.05), and the moderate or severe asthma group had significantly higher levels of IL-17 and TGF-β1 in serum and BALF than the mild asthma group (P<0.05). Levels of IL-17 and TGF-β1 in serum were significantly positively correlated with those in BALF (r=0.935 and 0.943, P<0.05 for both). In children with asthma, serum IL-17 level was significantly positively correlated with the percentage of neutrophils, eosinophils and epithelial cells in BALF (r=0.802, 0.799, and 0.674, P<0.05 for all), and a significant positive correlation was also seen between serum levels of IL-17 and TGF-β1 (r=0.878, P<0.05).

CONCLUSIONSLevels of IL-17 and TGF-β1 in serum and BALF are elevated in children with asthma. IL-17 and TGF-β1 may be involved in the occurrence and development of asthma, and they play important roles in asthma attack and aggravation.

Asthma ; immunology ; pathology ; Bronchoalveolar Lavage Fluid ; chemistry ; cytology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Interleukin-17 ; analysis ; blood ; physiology ; Male ; Transforming Growth Factor beta1 ; analysis ; blood ; physiology