Objective To investigate the correlation between bone mineral density(BMD)and the PXh haplotype combining estrogen receptor (ER) gene Xba Ⅰ , Pvu Ⅱ polymorphisms and osteocalcin gene Hind Ⅲ polymorphism in postmenopausal women.Methods In 307 subjects,the BMD of lumbar vertebrae and proximal femur were measured by dual energy X-ray absorptiometry and the Xba Ⅰ and Pvu Ⅱ polymorphisms of ER gene and the Hind Ⅲ potymorphism of osteocalcin gene were detected by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP).Results (1)The BMD of greater trochanter was significantly lower in XX genotype group than in xx genotype group ( P<0.05).The BMD of femoral neck, greater trochanter and Ward's triangle were lower in Xx genotype group[(0.695±0.087)g/cm2 , (0.592±0.106)g/cm2, (0.500±0.115) g/cm2] and X allele group[(0.697±0.088)g/cm2 , (0.594±0.105)g/cm2, (0.505±0.123)g/cm2] than in xx genotype group[(0.737±0.108) g/cm2,(0.653±0.119)g/cm2 ,(0.554±0.130)g/cM2] and non-X allele group[(0.737 ± 0.108) g/CM2, (0.653 ± 0.119) g/cm2 , (0.554 ± 0.130) g/cm2] ,respectively (all P<0.05 ).(2)The BMD of Ward's triangle was lower in PP genotype group and P allele group than in pp genotype group and non-P allele group (P<0.05).(3)The BMD of femoral neck, greater trochanter and Ward's triangle were lower in hh genotype group and h allele group than in I-IH genotype group, and were lower in non-h allele group than in HH genotype group(all P<0.05).(4)Women carrying PX, PXh haplotypes combining ER gene and osteocalcin gene had lower BMD at femoral neck than those not carrying PX,not carrying PXh haplotypes, respectively (all P<0.05).ConclusionsER gene(Xba Ⅰ) polymorphism and osteocalein gene(Hind Ⅲ) polymorphism are associated with BMD in postmenopausal women.The presence of X allele or h allele　shows negative influence on the BMD of postmenopausal women.The PXh haplotype is a suitable　genetic marker of postmenopausal women osteoporosis in Fuzhou area.

This study is to explore characteristic indexes in evaluation criteria for rat skin anaphylactoid test comparing skin blue spot OD values at the treated position and the control position in the same animal. Common contrast agents, traditional Chinese medicine injections and injections' active pharmaceutical ingredients or excipients in the existing clinical anaphylactoid reaction reports were taken as test drugs in the rat skin anaphylactoid test to define the K value: K > 2 represents positive anaphylactoid reaction, 1.2 ≤ K ≤ 2 represent doubtable anaphylactoid; K < 1.2 represents negative anaphylactoid reaction, which were taken as the criteria for evaluating anaphylactoid of tested drugs. The evaluation result and that for classic criteria were compared to study the applicability of K value. According to the comparison, K value, as the evaluation criteria in the rat skin anaphylactoid test, can more truly reflect the actual situation of skin aizen and minimize the error caused by animal individual factors. Compared with positive and negative two-level criteria for blue spot diameter, K value's positive, doubtable and negative three-level criteria are more objective and accurate. Therefore, K value can be used as the evaluation criteria in the rat skin anaphylactoid test.
Animals ; Drug Hypersensitivity ; immunology ; Drugs, Chinese Herbal ; adverse effects ; Female ; Humans ; Rats ; Rats, Sprague-Dawley ; Skin Tests ; methods

To obtain the recombinant pZP3alpha protein for the study of the contraceptive vaccines, the DNA sequence (446-1423) encoding purified pZP3alpha was inserted into a vector--pPICZalphaA. The recombinant plasmid pPICZalphaA-pZP3alpha was linearized and then transformed into Pichia pastoris GS115 by electroporation. Engineering strains were attained by screening with zeocin and induced to produce rpZP3alpha in high-density fermentation. Then rpZP3alpha was purified by Cu2+ metal affinity column chromatography from the separated and concentrated fermentative supernatants. The purified rpZP3alpha was identified by SDS-PAGE and Western blot, and the quantity, purity and rate of recovery of the rpZP3alpha were analyzed by Quantity One software. One male rabbit was immunized with the Cu-NTA-purified rpZP3alpha. The antibody responses against rpZP3alpha and porcine ZP were detected by ELISA and the indirect immunofluorescence. Engineering strains expressing rpZP3alpha in secretion were constructed. A 46kD component named rpZP3alpha which can react with anti-pZP3 antibody was purified from fermentative supernatants of engineering strains and the average yield of purified rpZP3alpha obtained from fermentative supernatants was 8mg/L. The purity and the rate of recovery were up to 92% and 63% respectively. The anti-rpZP3alpha antiserum was prepared by immunization of a male rabbit with purified rpZP3alpha. This anti-rpZP3alpha antiserum could react with rpZP3alpha and purified pZP3 in ELISA and bind to porcine zona pellucida which produced bright green fluorescence in the indirect immunofluorescence. The rpZP3alpha (46kD) protein could be successfully expressed in the Pichia pastoris expression system. And this protein retained the immunogenic activity of natural pZP3.
Animals ; Egg Proteins ; genetics ; metabolism ; Electroporation ; Fermentation ; Immunization ; Male ; Membrane Glycoproteins ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Rabbits ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; immunology ; secretion ; Swine ; Zona Pellucida Glycoproteins

OBJECTIVETo test the feasibility of correcting conjunctival sac narrowing following orbital implantation using polyester fiber heart patches instead of the skin autograft.

METHODSTwelve patients of conjunctival sac narrowing after orbital implantation (including 3 with orbital implant exposure) admitted in Nanfang Hospital between 2012 and 2016 received surgical correction of the conjunctival sac using polyester fiber heart patches. During the surgery, the central conjunctival sac was opened, the exposed area was covered with suitable polyester fiber heart patches, and the palpebral margin was sutured.

RESULTSThree months after the operation, 10 patients showed improved appearance after implantation of the prosthetic eye. Two patients received a second operation to remove the patches due to graft rejection and infections and skin autograft was implanted for reconstruction of the conjunctival sac.

CONCLUSIONPolyester fiber heart patches are ideal materials for repairing Conjunctival sac narrowing and orbital implant exposure, but this approach is not suitable in cases of severe narrowing or occlusion of the conjunctival sac.

Objective To investigate the association between interleukin-10 (IL-10) gene promoter microsatellite polymorphisms and the susceptibility to hepatitis B virus infection in Han,Yi and Yao ethnicities in GuiZhou province.Methods 500 volunteers were selected from Guizhou province.Ailelic frequency of IL-10.G and IL-10.R loci was identified by short tandom repeat polymerase chain reaction.The relativity between allelic frequency and HBV infection was analyzed.Results Genotype data from H-W analysis on all the IL-10 polymorphisms indicated that it was a random distribution.Very high HBV infection rates were found in the native ethnic minorities of Guizhou province.The overall HBV infection rate among the total population was 67.00％,with the HBV infection rates of Yi nationality in Weining,Yi nationality in Qianxi,Yao nationality in Libo and Han nationality in Libo as 51.85％,42.86％,79.52％ and 84.30％,respe~vely.The polymorphisms distribution of IL- 10.G and IL- 10.R were statistically different among the ethnic groups (P＜ 0.05 ).The polymorphisms distribution of IL-10.R had no significant difference between HBV infection group and non-infection group,as well as among HBV natural removal group and non-infected group in all the ethnic groups.The frequency of IL-10.G 459 bp (19CA) was significantly higher in non-infection group than in the infected group (P＜ 0.05 ).The frequency of IL-10.G 471 bp (25CA) was significantly higher in the non-infection group than in the HBV natural removal group(P＜0.05).The polymorphisms distribution of IL-10.G did not show significant difference between the HBV infection group and the HBV natural removal group in all the ethnic groups.We did not find any differences in allelic and genotypic frequencies of IL-10.G between infection group and non-infection group in Yi nationality in Weining,and Yao nationality in Libo (P＞0.05),as well as HBV natural removal group and non-infected group (P＞0.05).Conclusion The polymorphisms distribution of IL-10.R and IL-10.G did not show significant difference in Yi,Yao and Han ethnics population living in Guizhou province.IL-10.G seemed to influence the susceptibility of HBV infection in Han,Yao and Yi ethnics population of Guizhou province.

AIMTo study the effect of ligustrazine (LGT) and L-arginine(L-Arg)on function of mitochondria in myocardium after myocardial ischemia/reperfusion injury (MI/RI).

METHODS50 rabbits were randomly divided into five groups (n=10): Control group(A), MI/R group(B), MI/R + LGT group (C), MI/R+ L-Arg group (D), MI/R+ LGT + L-Arg group (E). The mitochondrial respiratory function, Ca2+ concentration ([Ca2+]m), malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were deter mined. Meanwhile, the contents of ATP and EC in the myocardial tissue were measured, respectively.

RESULTSIt was found that mitochondrial respiratory control rate (RCR), state 3 (ST3), SOD in C, D, E group were higher than those of B group, state 4 (ST4), [Ca2+]m, MDA were lower than those of B group, ATP and EC levels of myocardial tissue were higher than those in B group; and there were not significant differences between E and A group of above.

CONCLUSIONLGT and IL-Arg can improve function of mitochondria in myocardium after ischemia/reperfusion injury of myocardium in rabbits by decreasing oxygen free radical level and Ca" overload in the mitochondria.

Animals ; Arginine ; pharmacology ; Calcium ; metabolism ; Malondialdehyde ; analysis ; Mitochondria, Heart ; drug effects ; metabolism ; Myocardial Reperfusion Injury ; metabolism ; physiopathology ; Pyrazines ; pharmacology ; Rabbits ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the frequency of a 9 bp deletion polymorphism of mitochondrial DNA (mtDNA) in ethnic Miao, Buyi and Dong populations from Guizhou province.

METHODSPolymerase chain reaction-polyacrylamide gel electrophoresis (PCR-PAGE) was used to detect the 9 bp deletion. The result was verified with DNA sequencing.

RESULTSTwo polymorphisms, including a standard pattern and a short pattern (the 9 bp deletion), were found among the three ethnic groups. The frequency of short pattern in 304 males was 23.0%. Respectively, those of Miao, Buyi and Dong ethnics were 28.6%, 26.8% and 13.7%. A statistically significant difference was detected among the three groups (P<0.05).

CONCLUSIONThe frequencies of the 9 bp polymorphism were relatively high among ethnic Miao, Buyi and Dong populations from Guizhou, and there was a significant difference between the three.

Base Sequence ; China ; ethnology ; DNA, Mitochondrial ; genetics ; Gene Deletion ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Sequence Analysis, DNA

OBJECTIVETo evaluate the relationship of chimerism status of cell subsets with engraftment, occurrence of acute graft versus host disease (aGVHD), graft rejection and disease relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSChimerism status in peripheral blood (PB) and bone marrow (BM) of 65 patients received allo-HSCT were monitored at regular intervals post-transplant. Fluorescence-activated cell sorter (FACS) was used to sort CD3(+)T lymphocytes in 65 cases, CD3(-)CD56(+)CD16(+)NK cells in 52 cases, CD15(+) granulocytes in 32 cases and CD19(+)B lymphocytes in 20 cases post transplants. The chimerism status of different lineage cells was analyzed by polymerase chain reaction amplification of short tandem repeats (PCR-STR).

RESULTSOn day +7, NK-cells donor chimerism (DC 55.5%) was higher than other cell subsets. T lymphocyte was the latest one to reach complete donor chimerism (CDC) with a median on day +21. Patients whose T lymphocytes donor chimerism was more than 70% on day +7 and more than 95% on day +14 had a high risk for acute aGVHD. In all cases except those with ALL, the decreased DC of T lymphocytes were observed before molecular or hematological relapse occurred.

CONCLUSIONSerial and quantitative T cell chimerism analysis provides a reliable and rapid screening method for the early detection of engraftment, graft rejection, disease relapse and occurrence of aGVHD, therefore, is a prognostic tool to identify patients at high risk of aGVHD and disease relapse following allo-HSCT.

Adolescent ; Adult ; Child ; Chimerism ; Female ; Graft Rejection ; immunology ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Recurrence ; T-Lymphocytes ; immunology ; Young Adult

OBJECTIVETo compare the outcomes of intracytoplasmic sperm injection (ICSI) with retrieved epididymal and testicular sperm for obstructive azoospermia and with ejaculated sperm for severe oligozoospermia and asthenospermia.

METHODSWe retrospectively analyzed 431 ICSI cycles, which were divided according to sperm sources into Groups A (n=287 in patients with severe oligozoospermia or asthenospermia using ejaculated sperm), B (n=109 in obstructive azoospermia patients with sperm retrieved by percutaneous epididymal sperm aspiration, PESA) and C (n=35 in obstructive azoospermia patients with sperm retrieved by testicular sperm extraction, TESE). Comparisons were made among the three groups in the rates of embryo implantation, fertilization, pregnancy, cleavage, and miscarriage.

RESULTSGroup A showed statistically significant differences from Groups B and C in the rates of embryo implantation and pregnancy (18.46% vs. 25.23% and 28.76%, 31.23% vs. 42.16% and 39.39%, P < 0.05). But no significant differences were seen in the rates of fertilization, cleavage and miscarriage among the three groups (P > 0.05).

CONCLUSIONThe rates of embryo implantation and clinical pregnancy are higher in patients with obstructive azoospermia than in those with severe oligozoospermia or asthenospermia after ICSI with ejaculated sperm.

Azoospermia ; therapy ; Epididymis ; cytology ; physiopathology ; Female ; Humans ; Male ; Oligospermia ; therapy ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods ; Spermatozoa ; Testis ; cytology ; physiopathology

OBJECTIVETo investigate the variance and drug resistance of pathogenic bacteria isolated from children with infectious diseases seen between 2001 and 2006 in Chengdu area.

METHODSA total of 2888 pathogenic bacterial strains isolated from children in Chengdu Children's Hospital from 2001 to 2006 were analyzed. Tests were performed according to the guidelines of National Committee for Clinical Laboratory Standards (NCCLS) of the United States.

RESULT(1) Of the 2888 strains, 1845 (63.9%) were Gram negative bacteria. The main pathogenic bacteria included Escherichia coli (Ec, 718 strain, 24.9%), Hemophilus (H, 476 strain, 16.5%), Streptococcus pneumoniae (Sp, 412 strain, 14.3%), Klebsiella pneumoniae (Kp, 369 strain, 12.8%), Staphylococcus aureus (Sa, 353 strain, 12.2%), Staphylococcus epidermidis (Se, 278 strain, 9.6%), Pseudomonas aeruginosa (Pa, 146 strain, 5.1%) and other non-zymocyte (Onz, 136 strain, 4.7%). (2) The common pathogens found in blood specimen were 158 strain, which included Se (78 strain, 49.4%), Ec (23 strain, 14.6%), Kp (17 strain, 10.8%), Sa (14 strain, 8.9%), Onz (14 strain, 8.9%), Sp (7 strain, 4.4%) and Pa (5 strain, 3.2%). (3) The number of common pathogens isolated from patients with lower respiratory tract infection was 2018, including Ec (441 strains, 21.9%), H (430 strains, 21.3%), Sp (368 strains, 18.2%), Kp (253 strains, 12.5%), Sa (207 strains, 10.3%), Se (149 strains, 7.3%), Pa (97 strains, 4.8%) and Onz (73 strains, 3.6%). (4) There were 120 strains of common pathogens isolated from urine specimens, including Ec (78 strains, 65%), Kp (25 strains, 20.8%), Onz (7 strains, 5.8%), Pa (5 strains, 4.2%) and Se (5 strains, 4.2%). (5) There were 497 strains of common pathogens found in pus specimens, including Ec 167 strains, (33.6%), Sa (126 strains, 25.4%), Se (46 strains, 9.3%), H (44 strains, 8.9%), Onz (37 stains, 7.4%), Kp (31 strains, 6.2%), Sp (26 strains, 5.2%) and Pa (20 strain, 4.0%). The trend of drug resistance of pathogenic bacteria to antibiotics deteriorated. The proportion of methicillin-resistant Staphylococcus aureus (MRSA) was 6.7% and the methicillin-resistant coagulase-negative Staphylococci (MRCNS) rate was 20% in 2001 - 2003. The total proportion of extended-spectrum beta-lactamase stains (ESBL(S)) in Ec and Kp was 21.8%, and the rate of beta-lactamase production stains of Hemophilus influenzae (Hi) was 19.4% in 2001 - 2003.The proportion of MRSA was 17.2% and the MRCNS rate was 70.2%, the total proportion of ESBL(S) in Ec and Kp was 43.8%, and the rate of beta-lactamase producing stains of Hi was 39.7% in 2004 - 2006.

CONCLUSIONThe distribution of common pathogenic bacteria seen in Chengdu Children's Hospital has changed and the trend of drug resistance of pathogenic bacteria to antibiotics deteriorated in recent three years. Regionally monitoring the changes in pathogenic bacteria and the trend of drug resistance to antibiotics is paramount in guiding the pediatric clinical treatment.

Anti-Bacterial Agents ; pharmacology ; Bacterial Infections ; epidemiology ; microbiology ; Child ; China ; epidemiology ; Drug Resistance, Multiple, Bacterial ; drug effects ; Humans ; Microbial Sensitivity Tests