Aim To investigate the analgesic effect of tetrahydropalmatine on Cav1 . 2 expression in the dorsal root ganglion ( DRG) of mice with sciatic nerve chronic constriction injury ( CCI ) -induced neuropathic pain. Methods Forty male C57 BL/6 mice were randomly divided into 5 groups ( n =5 ): sham group ( group S) , CCI group ( group C ) and L-THP group ( group L) . Steady mice models of neuropathic pain were es-tablished by inducing CCI of sciatic nerve. According to development of neuropathic pain in mice, L group was divided into induction period, induction with ma-intenance period and long-term low-dose group. The mice were intraperitoneally administered with 45 mg · kg-1 tetrahydropalmatine in induction ( day 0~5 ) , in-duction with maintenance ( day 0~5 , 14~19 ) period of neuropathic pain state. From the instant after opera-tion, 15 mg · kg-1 tetrahydropalmatine was injected into the long-term low-dose group once per day for 19 days. Then, the behavior changes of mice were moni-tored. Moreover, the threshold of mechanical and ther-mal stimuli was tested. In addition, the expression of Cav1 . 2 protein was detected by Western blot and im-munohistochemical staining. Results The lowest ex-pression of Cav1 . 2 was observed in group C and the highest expression level of Cav1 . 2 was found in group S. Cav1. 2 expression was significantly up-regulated in induction period group, induction with maintenance period group and long-term low-dose group ( P<0. 05 , P<0. 01). Compared with group C, high dose of tet-rahydropalmatine in induction period group, induction with maintenance period group and long-term low-dose group showed reduced mechanical allodynia and ther-mal hyperalgesia induced by nerve injury ( P <0. 05 , P<0. 01). Meanwhile, high dose of tetrahydropalma-tine significantly relieved the mechanical allodynia in induction period group, induction with maintenance period group and thermal hyperalgesia in maintenance period group (P<0. 05). However, there was no ob-vious effect on mechanical allodynia and thermal hyper-algesia induced by nerve injury ( P >0. 05 ) in long-term low-dose group. Conclusions High dose of tet-rahydropalmatine in induction period group, induction with maintenance period group and low-dose among the whole experiment process obviously relieves the neuro-pathic pain induced by nerve injury. The analgesic effect of tetrahydropalmatine on neuropathic pain may be due to the increased expression of Cav1 . 2 protein in DRG neurons.

We have studied the proteomic changes of the serum of the Smad3 targeted deficient mice using 2-DE and PMF approaches. 7 proteins expressed at different level in wild type mice and the Smad3 deficient mice were identified. These results would benefit the research on diagnosis and therapy of osteoarthritis and provided clues to studying the important function of Smad3 mediated TGF-beta signals during the skeletal development.
Animals ; DNA-Binding Proteins ; blood ; deficiency ; genetics ; Electrophoresis, Gel, Two-Dimensional ; Electrophoresis, Polyacrylamide Gel ; Genotype ; Mice ; Mice, Knockout ; Peptide Mapping ; Proteome ; analysis ; Smad3 Protein ; Trans-Activators ; blood ; deficiency ; genetics

OBJECTIVETo screen the severe acute respiratory syndromes (SARS) mimotopes with random phage peptide library and to investigate their immunogenicity.

METHODSUsing SARS sera as selective molecule, a 12 mer phage peptide library was biopanned and positive clones containing the mimic epitopes were selected. The immuno-characteriation of the epitopes were then investigated.

RESULTS2 positive clones that having specific affinity to SARS sera were obtained. The DNA sequencing data showed no homology between the sequences of the deduced amino acid of the two mimic antigen peptides and the sequence of SARS.

CONCLUSIONSARS mimotopes were obtained by phage peptide library screening. This method might provide a new approach for SARS therapy and vaccine development.

Animals ; Antigens, Viral ; Base Sequence ; Biomimetic Materials ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; genetics ; immunology ; Humans ; Peptide Library ; Sequence Analysis, DNA ; Severe Acute Respiratory Syndrome ; immunology

OBJECTIVETo observe the clinical effect of Yangyin Yiqi Huoxue Recipe (YYHR, the basic recipe of Yangyin Tongnao Granule) in treatment of ischemic stroke patients of deficiency of qi and yin syndrome (DQYS) and static blood obstructing collaterals syndrome (SBOCS).

METHODSTotally 312 patients were assigned to the control group (86 cases) and the treatment group (226 cases) using strati- fied randomized allocation method. Patients in the treatment group were treated with modified YYHR, while those in the control group took Xueshuan Xinmaining. The treatment course was 4 weeks for all. Constituent ratios of the acute stage and the recovery stage of DQYS and SBOCS and their complicated syndromes were observed in the two groups. Changes of the clinical curative effect, clinical symptoms integral, whole blood viscosity ratio, plasma viscosity ratio, hematocrit, erythrocyte sedimentation rate (ESR), total cho- lesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C) were detected in the two groups before and after treatment.

RESULTSThere was statistical difference in constituent ratios of the acute stage and the recovery stage of DQYS SBOCS and its complicated syndromes between the two groups (P < 0.01). DQYS and SBOCS was basic syndrome types of the two groups. The cured and markedly effective rate was 71.24%(161/226) in the treatment group and 43.02% (37/86) in the control group. The total effective rate was 91.15% (206/226) in the treatment group, higher than that of the control group (76.74%, 66/86) with statistical difference (P < 0.01). There was statistical difference in the clinical symptoms integral, whole blood viscosity ratio, plasma viscosity ratio, hematocrit, ESR, TC, TG,HDL-C, and LDL-C (P < 0.05, P < 0.01).

CONCLUSIONSSymptoms of ischemic stroke patients could be improved by modified YYHR. Indices such as the whole blood viscosity, plasma viscosity ratio, hematocrit, ESR, abnormal metabolism of blood lipids were also significantly improved. Pathological changes of blood stasis induced by qi-yin deficiency exist in ischemic stroke patients, and DQYS and SBOCS were basic syndrome types.

Aged ; Biomedical Research ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hematocrit ; Humans ; Male ; Middle Aged ; Qi ; Stroke ; drug therapy ; Triglycerides ; blood ; Yin Deficiency ; therapy

AIMTo evaluate the effect of intracytoplasmic sperm injection (ICSI) in the management of cases with a history of conventional in vitro fertilization (IVF) failure.

METHODSTwo groups of patients, 19 with normal semen parameters and a history of IVF failure (metaphase II oocytes: 0-30 %) and 28 with severe male factor infertility received ICSI technology during the same period. Ovarian stimulation was achieved by conventional procedure. Transvaginal ultrasound-guided oocyte collection was done 35-37 h after human chorionic gonadotrophin (hCG) injection. Only metaphase II oocytes were selected for microinjection.

RESULTSFertilization was achieved with ICSI in all the patients. The fertilization rate (75.6 % +/-21.1 % vs. 73.9 % +/-19.2 %), cleavage rate (85.1 % +/-19.3 % vs. 82.7 % +/-22.1 %), clinical pregnancy rate per embryo transfer cycle (31.6 % vs. 28.6 %) and implantation rate per embryo (15.3 % vs. 14.4 %) did not differ significantly between the two groups.

CONCLUSIONICSI is a valuable method for couples with a history of IVF failure. These patients may have a similar ICSI result as in severe male infertility.

Abortion, Spontaneous ; Adult ; Female ; Fertilization in Vitro ; methods ; Humans ; Infertility, Male ; therapy ; Male ; Oocytes ; Pregnancy ; Pregnancy Rate ; Pregnancy, Ectopic ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; Treatment Failure

We determine the contamination and genotypes of Clostridium perfringens isolated from chickens in Macau.C. perfringens were isolated from these samples by double tube method and then identified by biochemical test.Genotyping of i-solated strains was determined by multiplex PCR.A total of 120 samples,the positive rates were 37.5%,respectively.All of the isolated strains were type A.This study indicated that contamination of C.perfringens type A exists in chicken intestines, fresh chickens and chilled chickens in Macau,and provided reference data for the public health problems caused by C.perfrin-gens.

Objective To evaluate the diagnostic advantage of the ratio of ligamentum flavum(LF) thickness to oblique canal diameter(TODR)measured on CT images in patients with lumbar canal stenosis. Methods Seventy-one patients underwent CT and MRI examinations respectively,and they were divided into two groups,the positive group and negative group,according to the presence or absence of dural sac notch caused by the LF on bilateral parasagittal MR images.Meanwhile,50 volunteers without any symptom in the lumbar region or legs were examined by CT.TODRs were measured at the L3—S1 levels of the inferior margin of the intervertebral disc on transverse CT images,respectively.The results were further analyzed with the positive findings on MR images,clinical symptoms and physical examination,so as to find out the statistical correlation between them.Results LF thickness was(3.01?0.72)mm and TODR was 0.19?0.04 in the negative group,(3.94?0.84)mm and 0.28+0.06 in the positive group,and(3.16? 0.85)mm and 0.19?0.04 in the control group.There was significant difference between positive group and negative group or control group for LF thickness(P0.24, the sensitivity,specificity and positive predictive value were 74.8%,89.6% and 73.6% respectively. Positive correlation existed between LF thickness or TODR and clinical symptom(r=0.72,0.86,P

OBJECTIVETo investigate the association between a -799C/T polymorphism in the promotor region of matrix metalloproteinase-8 (MMP-8) gene and instability of carotid plaque in Chinese Han population.

METHODSA total of 451 acute infarction patients from the Department of Neurology of Taizhou Hospital were divided into carotid vulnerable plaque group and carotid stable plaque group according to the results of carotid B-mode ultrasonography. Serum MMP -8 level was measured by the means of enzyme-linked immunosorbent assay (ELISA). At the same time, the MMP-8 -799C/T polymorphism was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.

RESULTSSerum level of MMP-8 in the carotid vulnerable plaque group was higher than that in the carotid stable plaque group (t= 2.894, P= 0.004). The genotype distribution of -799C/T polymorphism between the two groups was significantly different (Chi-square = 13.65, P= 0.000). Serum level of MMP-8 in patients with TT genotype was higher than that in patients with CC genotype (t= 3.141, P= 0.001).

CONCLUSIONThe present study suggested that serum level of MMP-8 and the -799C/T polymorphism of MMP-8 gene are associated with carotid vulnerable plaque in Chinese Han population, and the T allele may be a predictor for the susceptibility of carotid vulnerable plaque.

Aged ; Base Sequence ; Female ; Genotype ; Humans ; Male ; Matrix Metalloproteinase 8 ; genetics ; Molecular Sequence Data ; Plaque, Atherosclerotic ; enzymology ; genetics ; pathology ; Polymorphism, Genetic ; Promoter Regions, Genetic

OBJECTIVETo investigative vacA, cagA and iceA genes dominant genotypes of Helicobacter pylori (Hp) isolated from children suffering from gastric and duodenal diseases in Guangzhou area.

METHODSTotally 105 children who underwent gastroscopy in Guangzhou Children's Hospital were enrolled into this study. From each patient, 3 biopsy specimens from the gastric antrum were taken, one was used for rapid urease test, one for histological examination, and one for polymerase chain reaction (PCR) for detecting ureA, vacA, cagA, and iceA genes. DNA was prepared directly from the biopsy specimens from the gastric antrum using a QIAamp DNA mini kit (Qiagen, Germany) according to the manufacturer's instructions. Then 11 primers were used for detecting the genotypes including ureas, (s1, s1a, s1b, s1c, s2) and m (m1, m1T, m2) region of vacA, cagA and iceA (iceA1 and iceA2) genotypes in the 105 children. The distribution of the genotypes of Hp was analyzed.

RESULTAmong the 105 children, only 52 children were positive by the three methods, among these 52 children, 26 were boys and 26 girls. Hp vacA s1as1c/m2 was detected in 43 out of 52 children (82.7%), s1as1c/m1T in 9.6% (5/52), m region that could not betyped was 7.7% (4/52). No strains presented genotypes vacA s1b, s2, m1. The comparison of the positive ratio of vacA s1as1 c/m2 detected in the children infected with Hp and that of the other combination of signal region and middle region was statistically significantly different (P < 0.01). With regard to cagA gene, cagA(+) gene and cagA(-) gene were found in 90.4% (47/52) and 9.6% (5/52) of the children, respectively. The cagA(+) gene was more frequent in the children infected with Hp. Single iceA1 was detected in 78.8% (41/52) children, and single iceA2 was detected to be 1.9% (1/52), multiple strains infection of iceA1 and iceA2 were detected in 3.8% (2/52) children, iceA1 and iceA2 were not detected in 15.4% (8/52), the comparison of the positive ratio of iceA1 detected in the children infected with Hp and that of the other genotypes was statistically significantly different (P < 0.01).

CONCLUSIONThe s1as1c/m2, cagA and iceA1 were the dominant genotypes of Hp in the children in Guangzhou area and s1as1c/m2, cagA and iceA1 were the dominant genotypes combination of Hp in the children in this area.

Antigens, Bacterial ; genetics ; therapeutic use ; Bacterial Outer Membrane Proteins ; genetics ; Bacterial Proteins ; genetics ; Child ; China ; epidemiology ; Female ; Genes, Bacterial ; drug effects ; genetics ; Genotype ; Helicobacter Infections ; drug therapy ; genetics ; Helicobacter pylori ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; Pyloric Antrum ; microbiology

The aim of this study was to analyze chimerism, evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) by multiple short tandem repeat (STR) amplification using fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis. Peripheral blood and bone marrow in 27 patients who received myeloablative allogenetic cell transplantation were collected before and after transplantation in different times. 10 and 7 different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus/Cofiler plus PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI prism 310 Genetic Analyzer with capillary electrophoresis. The Genescan and Genotype soft ware were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on the different allelic distribution type between donor and recipient. The results showed that donor chimerism was similar by the two methods. The median number of informative alleles was 6.3 (4 - 9) by Profiler Plus and 4.9 (2 - 6) by Cofiler Plus. The donor alleles appeared in 26 patients on day 28 post transplantation. One patient was not observed to appear donor alleles. 14 patients with 100% donor chimerism (DC) had stable engraftment and they still survive in free leukemia. 9 patients had unstable mixed chimerism (DC: 0% - 90.2%), and 5 of them relapsed after allo-HSCT, 6 patients died. Decrease of donor chimerism appeared prior to graft rejection and disease relapse. The incidence of GVHD was higher in group of full donor chimerism. It is concluded that dynamic monitoring donor chimerism by STR-PCR in combination with all auto-capillary electrophoresis is a valuable tool for predicting graft rejection, disease relapse and occurrence of GVHD, and provides a basis for early clinical intervention in the patients received allo-HSCT.
Adolescent ; Adult ; Female ; Graft vs Host Disease ; prevention & control ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Male ; Peripheral Blood Stem Cell Transplantation ; methods ; Polymerase Chain Reaction ; methods ; Recurrence ; Tandem Repeat Sequences ; Transplantation Chimera ; Transplantation, Homologous