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.

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.

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 investigate the expression of VEZT and its clinical pathological significance in gastric cancer tissues,and to construct the VEZT over-expression vector.Methods The expression of VEZT was examined in 119 cases of gastric carcinoma and their corresponding normal mucus tissues by SP immunohistochemical staining.The relationships between the VEZT expression levels and its clinicopathological characteristics,prognosis were also investigated.VEZT cDNA was extracted and amplificated from 293 cells by polymerase chain reaction (PCR).Using DNA recombinant technique,the target gene was connected to the pEGFP-N1 vector to construct recombinant plasmid vector after the target gene and pEGFP-N1 were purified.The recombinant plasmid was identificated by 1％ enzyme electrophoresis and DNA sequencing.Results The VEZT-positive expression in gastric carcinoma was 30.3％ and 60.5％ in normal gastric tissues.VEZT expression in high differentiated cancer tissues was higher than that in lower differentiated tissues(x2 =5.002,P ＜ 0.05),expression of VEZT in early gastric cancer was significantly higher than that in patients with advanced gastric cancer (x2 =5.551,P ＜ 0.05),expression of VEZT in patients without lymph node metastasis was significantly higher than that of lymph node metastasis (x2 =5.878,P ＜ 0.05).Patients with positive VEZT expression have better prognosis than the negative patients(x2 =6.908,P ＜ 0.01).The target fragment,consistent with the theoretical value,was verified by gel electrophoresis.DNA sequencing confirmed that the gene sequence had no mutation and the eukaryotic expression plasmid vector pEGFP-N1-hVEZT was successfully constructed.Conclusions VEZT protein was correlated with TNM staging,tumor stage,invasion depth and lymph node metastasis,positive VEZT expression predicting better five year survival.The VEZT over-expression plasmid vector was successfully constructed.

Objective To analyze the correlation between mutation in JAK2-V617F gene in peripheral blood mononuclear cells and thrombotic events in patients with myeloproliferative.Methods Investigation of thrombosis in 391 patients with MPN,using QRT-PCR to detect JAK2-V617F gene mutation in peripheral blood of patients.Result 105 cases of thrombotic events in 391 MPN patients (26.9％),84 cases of thrombotic events in 273 JAK2-V617F gene mutation positive patients (30.8 ％),however,21 cases of thrombotic events in 118 J AK2-V617F gene mutation negative panents (17.8 ％).The difference between the two groups was statistically significant (x2 =32.30,P＜0.01).In the PV group,45 cases of thrombosis events in JAK2-V617F positive patients (29.2 ％),6 cases of thrombosis events in JAK2-V617F negativepatients (42.9％),there was significant difference between the two groups (x2 =2.13,P＞0.05).In the ET group,18 cases of thrombosis events in JAK2-V617F positive patients (32.2％),9 cases of thrombosis events in JAK2-V617F negative patients (15.8％),there was significant difference between the two groups (x2 =12.32,P＜0.01).In the PMF group,21 cases of thrombosis events in JAK2-V617F posinve patients (33.3 ％),6 cases of thrombosis events in JAK2-V617F negativepatients (12.8 ％),there was significant difference between the two groups (x2 =14.32,P＜0.01).Conclusion JAK2-V617F positive MPN patients were more susceptible to thrombotic events especially in patients with ET and PMF.

Objective To evaluate the therapeutic effects of ultrasound biomicroscopy (UBM) assisted intraocular lens suspension surgery with one loop suturing for residual lens capsule aphakia.Methods Fifty aphakia patients (50 eyes) with residual lens capsular equal or greater than 180 degrees were chosen.The patients were randomly divided into 2 groups and underwent intraocular lens suspension surgery with one loop suturing:Trial group (25 cases,25 eyes) selected suture site according to the preoperative UBM results,and control group (25 cases,25 eyes) selected the suture site according to the clinical experience.The uncorrected visual acuity (UCVA),astigmatism at preoperative and postoperative 1 week,1 month and 3 months were observed,the eccentric and tilt value of the lens were calculated by UBM measurement at 3 month after operation,and the postoperative complications of the two groups were observed.Results UCVA at each time point after operation were higher than that before operation,and gradually increased with time.There was no significant difference between the two groups at 1 week before and after the operation of UCVA (P ＞ 0.05).At 1 month and 3 months after operation,UCVA in trial group were significantly higher than those of control group (all P ＜ 0.05).The astigmatism value at 1 week after operation in two groups was higher than that before operation and gradually decreased with time.The astigmatism value in trial group at 1 month and 3 months after operation had no significant difference with that of preoperative (P ＞ 0.05),while the astigmatism value in control group increased (all P ＜ 0.05).The astigmatism values had no significant difference between the two groups before and 1 week after operation (all P ＞0.05),which were significantly lower in trial group at 1 month and 3 months after operation (all P ＜ 0.05).Three months after surgery,the proportion of lens accurately implanted in the ciliary sulcus was significantly higher than control group (P ＜ 0.01),the eccentric and tilt values of the lens in trial group were significantly less than control group (P ＜ 0.01).No serious complication was observed in all patients.Conclusion UBM assisted intraocular lens suspension surgery with one loop suturing can significantly improve the postoperative visual quality,safety and lens stability,which can be used as a routine preoperative examination of intraocular lens suspension surgery.In addition,UBM provides an alternative method for the calculation of the eccentricity and tile value of the intraocular lens after the implantation of the intraocular lens.

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

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.