Objective:To prepare and purify recombinant human zona pellucida 3 protein,and to study its immunologic activity.Methods:The E.coli BL21 containing recombinant plasmid pGEX4T-1/ huZP3 was induced to express GST-fusion protein by IPTG.After a series of purification procedure,the purified protein was analyzed by SDS-PAGE.After the mice immunized with rhuZP3,the antibody responses against rhuZP3 were detected by ELISA.Results:The soluble fusion protein was expressed,and purity of rhuZP3 was 95%.Moreover, purity of rhuZP3 could be recognized by anti-human ZP3 in ELISA.Conclusion:The rhuZP3 is obtained through the preparation of prokaryotic expression system and anti-rhuZP3 antibody has immunological activity.

At the urgent request of Artemisia annua (ART) planting, the paper gets artemisinin content (ARTC) of ART in China from literatures. The paper analyses the relationships between ARTC and ecological factors by statistical analytical methods. The paper also analyses the climate suitable rank distribution of ARTC in China by ArcGIS. The results display that first, ARTC is significantly different in China, that ART from the south regions ARTC is higher. Greatest north parts of China have not suitable climate for the growing of ART and the ARTC is lower than 0.2%, when ART grows above the 34th degree of northern latitude. ARTC is higher and ART grows well, when ART grows under the 34 degrees N and grows at the areas between 100 degrees E and 120 degrees E. Second, subtropical zone is the best suitable climate zone for the growing of ART. ART grows well and ARTC is higher than 0.5%, when ART grows in the subtropical zone. Third, temperature, sunshine duration and rainfall are the main ecological factors that affect the growth of ART and the accumulation of ARTC. That the year temperature between 13.9 degrees C and 22 degrees C, sunshine duration between 853 h and 1507 h, rainfall between 814 mm and 1518 mm, is the best climate for the accumulation of ARTC. Temperature between 13 degrees C and 29 degrees C, rainfall between 600 mm and 1300 mm is the best climate for the growth of ART. Fourth, in northwest of Guangxi, eastern of Sichuan, Guizhou and Yunnan provinces, south Chongqing and west Hunan Province, there are suitable climate for the growth of Artemisia and for the accumulating of ARTC. There are also some suitable climate areas for the growing of artemisia in the south of Hubei, Anhui and Jiangsu provinces.

OBJECTIVETo explore the incidence, degree, and pattern of rib cartilage calcification in congenital microtia patients, in order to provide reference for harvesting the rib cartilage, sculpturing cartilage framework.

METHODSFrom Jun. 2013 to Nov. 2014, 383 patients (age range, 6-45 years) underwent CT scans of the chest. 11 patients with bony diseases or traumatic history were excluded. The remaining 372 patients were divided by age into four groups as 6-15, 16-25, 26-35, 36-45 years old. Twenty patients (10 male and 10 female) were selected by the order of patient identification number in each age group, thus selecting a total of 80 patients (40 male and 40 female). Retrospective study of CT scans of the chest in 80 patients and the incidence, degree, and pattern of cartilage calcification of the sixth to eighth ribs were noted. A chi-square test is conducted to test whether there are significant difference between the variables through the SPSS 19.0 software.

RESULTSOverall, 40.4% (194/480) cartilage was calcified; female patients (47.50%, 114/240) showed higher frequency of calcification than male patients (33.33%, 80/240, P = 0. 025). Calcification rates of all age groups are 1.7% (2/120), 46.7% (56/ 120), 49.2% (59/120), 64.2% (77/120). Calcification rate of 6-15 years group is lowest in all groups (P < 0.05) while other three groups have no statistical significance (P > 0.05). Calcification rates of the sixth and sevent rib cartilage were higher than those of the eighth rib cartilage in all age groups except 6-15 years group, who had a similar rate of all three ribs. Calcification rate of all three rib cartilage was significantly increased with age. Calcification rates of the amle's rib cartilage and the female's in all age groups are 3.3% (2/60) and 0.0% (0/60) (6-15 years): 33.3% (20/60) and 60.0% (36/60) (16-25 years): 40.0% (24/60) and 58.3% (35/60) (26-35 years), 56.7% (34/60) and 71.2% (43/60) (36-45 years). In 6-15 years group calcification rates of male and female had a similar rate, while female's rates were higher than male's rates in other three groups. Male and females mainly had the granular type of calcification [70.0% (56/80), 63.2% (72/114)].

CONCLUSIONSFemales who are over 16 years old should pay more attention to the possibility of middle-severe calcification before harvesting rib cartilage. These patients should take CT examination if necessary. In addition, the patients who had previous operation, or traumatic history, rib deformity, or spine deformity should select the CT examination.

Adolescent ; Adult ; Age Factors ; Calcinosis ; diagnostic imaging ; epidemiology ; Cartilage Diseases ; diagnostic imaging ; epidemiology ; Child ; Congenital Microtia ; Costal Cartilage ; diagnostic imaging ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Retrospective Studies ; Sex Factors ; Tomography, X-Ray Computed

Objective To explore the effects of H.pylori and crude extracted proteins secreted by H.pylori(broth culture filtrate protein,BCF-P)on acid secretion from isolated rabbit parietal cells.Methods Parietal cells from rabbit gastric mucosa were isolated and enriched with digestion and elutriation.H.pylori(NCTC 11637,CagA+ VacA+)were grown in liquid broth culture and BCF-P was precipitated with ammonium sulfate.The vacuolation activity of BCF-P was evaluated with neutral red dye uptake test in HeLa cell.Isolated parietal cells were incubated with H.pylori(bacteria/cell=100∶1)for 2 h and 16 h,or BCF-P(100μg/ml)for 1 h and 12 h.Acid secretion from parietal cells was studied using 14C-aminopyrine(14C-AP)accumulation indirectly and H+-K+ ATPase α subunit mRNA expression was assessed using RT-PCR.Results (1)BCF-P containing vacuolating cytotoxin(VacA)with vacuolation activity on HeLa cells had positive result on neutral red uptake test.(2)The basal expression of H+-K+ ATPase α subunit mRNA could be detected in isolated parietal cells and 14C-AP accumulation was significantly increased in response to the stimulation of histamine with different concentrations for 30 min(P＜0.05).These results indicated that the isolated parietal cells retain relative intact acid secretion function.(3)The histamine(1.0×104 mol/L)stimulated acid secretion was inhibited sustainedly in response to H.pylori by 81% at 2 h and by 94% at 16 h(P＜0.05).However,H+-K+ ATPase α subunit mRNA expression was up-regulated in tlle acute period(2 h)and was down-regulated in the chronic period (16 h)by H.pylori(P＜0.05).(4)BCF-P significantly inhibited the histamine-stimulated acid secretion by 24% at 1 h and by 58% at 12 h(P＜0.05),and this inhibition was accompanied by the down-regulated expression of H+-K+ATPase α subunit mRNA.Conclusions Intact H.pylori and VacA secreted by H.pylori could directly inhibit histamine-stimulated acid secretion from parietal cells and this inhibition may be mediated by the down-regulated H+-K+ ATPase expression.

BACKGROUND:Surgery has become gold standard to treat fracture of acetabulum.The incidence of posterior column fracture of affected acetabulum is high.The appropriate treatment for posterior column fracture of acetabulum remains controversial.OBJECTIVE:To establish the model of posterior column fracture of acetabulum,and evaluate the stability of internal fixation with the plate and lag screw for the posterior column fracture of acetabulum.METHODS:A total of 20 preserved cadaveric semipelvic specimens were randomly divided into two groups.Models of isolated posterior column fracture of acetabulum were established.The models were fixed with plate or lag screw.Under vertical compression load,the horizontal displacements of the fracture site were measured and the shear rigidity of two fixation conditions was compared.RESULTS AND CONCLUSION:The horizontal displacements of the fracture site fixed with plate and lag screw were (1.64±0.17) mm and (1.70±0.20) mm.The shear rigidity fixed with plate and lag screw were (392.40±41.22) N/mm and (386.33±50.07) N/mm.The differences of both horizontal displacements and shear rigidity between the two groups were not significant (P>0.05).The fracture model provides an ideal tool for biomechanical evaluation of the stability of internal fixation for the posterior column fracture of acetabulum.There was no significant difference in the stability of internal fixation with the plate and lag screw for the posterior column fracture of acetabulum.

Objective To investigate the potential mechanism of interleukin-1β(IL-1β)in H.pylori-related gastric carcinogenesis.Methods Human gastric cancer cell line(AGS),human gastric epithelial cell line(GES-1)and isolated parietal cells were treated with exogenous IL-1β(10 ng/ml)in the presence or absence of H.pylori(NCTC 11637).Cell proliferation and apoptosis were determined by MTT assay and flow cytometry,respectively. Expressions of cyclooxygenase-2(COX-2)mRNA and protein were examined using RT-PCR and flow cytometry,respectively.Acid secretion by parietal cells was tested using 14C-aminopyrine(14 C-AP) accumulation indirectly.H+/K+ ATPase α subunit mRNA expression was assessed using RT-PCR.Results The cell proliferation and H.pylori-related apoptosis in both GES-1 and AGS celL lines were stimulated and inhibited by IL-1β.IL-1β induced expression and upregulation of COX-2 mRNA in GES-1 and AGS cell lines,respectively.In addition,IL-1β continuously inhibited the ability of histamine-stimulated 14C-AP accumulation of isolated parietal cells accompanied by down-regulation of H+/K+ ATPase mRNA expression.Conclusions It suggests that IL-1β play an important role in H.pylori-related gastric carcinogenesis through two pathways:①to interfere gastric epithelial cell growth by up-regulating COX-2 expression,②to inhibit acid secretion from parietal cell by down-regulating H+/K+ ATPase expression.

Objective To assess the treatment of fresh Monteggia fracture with a steel-wire loop for adults whose annular ligament needs repairing. Methods From May 2006 to March 2008, we treated 15 adult patients with fresh Monteggia fracture whose annular ligament needed repairing with a steel-wire loop. A retrospective study was performed for the 15 cases, including 9 males and 6 females. Their ages ranged from 19 to 46 years. According to the Bado classification system, there were 6 cases of type Ⅰ, 4 type Ⅱ, 5 type Ⅲ. The ulnar fractures were treated with open reduction and compression plate fixation. Next, radial head dislocations were reduced openly and the reduced heads were hitched to the proximal end of the ulna with a steel-wire loop of 0.8 mm in diameter around the radial neck. Last, the broken annular ligaments were repaired. The elbow functional trainings were carried out 48 hours after operation and the steel-wire was removed 3 weeks postoperatively. Results The patients were followed up for 8 to 11 months (mean,9. 5 months). The functional recovery evaluated by Broberg and Morrey evaluation system showed excellent results in 11 patients, good in 2, fair in 2 and poor in 0. The excellent to good rate was 86. 7%.Conclusion The steel-wire loop treatment is a good strategy for adults with fresh Monteggia fracture whose annular ligament needs repairing.

Objective: To establish the determination of adenosine in Ganping Capsules. Methods: HPLC method was developed for the determination of adenosine in Ganping Capsules on Kromasil C 18 (150mm? 4.6mm i.d.) in 0.05mol/L KH 2PO 4 methanol (88∶12, V/V,pH 4.7) as a mobile phase (1.0mL/min). The UV detection wavelength was set at 260nm. Adenosine was extracted with 10% methanol water. Results: The recoveries was 98%～102% and the linear equation was Y=2537.4X+100.81. The correlation coefficient was r =0.9999(n=5). Conclusion: The method is simple, rapid and accurate.

Objective To evaluate whether the comparability of 3 automatic blood cell analyzers meet the clinical requirements by conducting the comparative study on the detection results of these instruments.Methods With the Sysmex 2100 automatic blood cell analyzer as the reference instrument,Sysmex 1000i and Abbott 1800 as the experimental instrument,the original quality control provided by the instrument factory and the patient′s fresh anticoagulant blood samples in the laboratory were adopted to monitor for continuous 40 d by these three instruments and the detection results of WBC,RBC,HGB,HCT and PLT were analyzed.Results The detection results of these 3 instruments were statistically tested by the F test,the differences showed no statistical significance (P >0.05)and the bias was in 1/2 of the maximum permissible error range in America department clinical test revised regulations (CLIA′88).Conclusion The detection results by these 3 instruments are comparable and can meet the clinical requirements.