Bronchi ; cytology ; Cell Culture Techniques ; Cells, Cultured ; Humans ; Myocytes, Smooth Muscle ; cytology

Objective To construct and express the recombinant plasmid pET28α-Sj26GST-Sj32 of Schistosoma japonicum(Sj) in Escherichia coli BL21 (DE3). Methods Total RNA was extracted from Sj adult worms by ultrasound-breaking, Sj26GST and Sj32 antigen gene was respectively amplified by RT-PCR from the total RNA; Sj26GST-Sj32 fusion gene obtained with gene splicing by overlap extension(SOEing) was cloned into prokaryotic expression plasmid pET28α and transformed into Escherichia coli BL2 (DE3) to construct pET28α-Sj26GST-Sj32;BL21 (pET28α-Sj26GST-Sj32) was induced with isopropyl-β-D-thiogalactopyranosid (IPTG), and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE)and Western blotting. Results The 1991 bp Sj26GST-Sj32 fusion gene was successfully amplified by gene SOEing and cloned into pET28α by restriction analysis and PCR identification, the recombinant plasmid pET28α-Sj26GST-Sj32 was successfully constructed; the relative molecular mass of the expressed recombinant protein was approximately 69 × 103 by SDS-PAGE, and the amount of the expressed protein was 25% of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pET28α-Sj26GST-Sj32 is successfully constructed and highly expressed in Escherichia coli in fused form with His-tag, and the expressed fusion protein shows specific antigenicity.

Objective To construct and express the recombinant plasmid pET32α-Sj26GST of Schistosoma japonicum(sj)in Escherichia coli(E.coli)B121(DE3).Methods The total RNA was extracted from sj adult worms by ultrasound-breaking,Sj26GST antigen gene was amplified by RT-PCR from the total RNA,then cloned into prokaryotic expression plasmid pET32α(+) and transformed into E.coli B12(DE3)to construct pET32α-Sj26GST;BL21(pET32α-Sj26GST)WaS induced with isopropyl-β-D-thiogalactopyranosid(IPTG),and the expressed products were analyzed and identified by SDS-PAGE and Western blot.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and cloned into pET32α(+)by restriction analysis and PCR identification,the recombinant plasmid pET32α-Sj26GST was successfully constructed;the relative molecular mass of the expressed recombinant protein was approximately 49×103 by SDS-PAGE,and the amount of the expressed protein was 24%of the total bacterial proteins;the fusion protein could be recognized by sera from rabbits infected with sj by Western blot.Conclusions The recombinant plasmid pET32α-Sj26GST is successfully constructed and highly expressed in E.coli in fused form with Trx-tag and His-tag,and the expressed fusion protein shows specific antigenicity.

So far there exists no corresponding quality test procedures and grading standards for the seed of Prunus humilis, which is one of the important source of base of semen pruni. Therefor we set up test procedures that are adapt to characteristics of the P. humilis seed through the study of the test of sampling, seed purity, thousand-grain weight, seed moisture, seed viability and germination percentage. 50 cases of seed specimens of P. humilis tested. The related data were analyzed by cluster analysis. Through this research, the seed quality test procedure was developed, and the seed quality grading standard was formulated. The seed quality of each grade should meet the following requirements: for first grade seeds, germination percentage ≥ 68%, thousand-grain weight 383 g, purity ≥ 93%, seed moisture ≤ 5%; for second grade seeds, germination percentage ≥ 26%, thousand-grain weight ≥ 266 g, purity ≥ 73%, seed moisture ≤9%; for third grade seeds, germination percentage ≥ 10%, purity ≥ 50%, thousand-grain weight ≥ 08 g, seed moisture ≤ 13%.
Cluster Analysis ; Germination ; Prunus ; growth & development ; Seeds ; physiology

ObjectiveTo introduction of perforator flaps,muscle flaps pedicled by descending branch of lateral circumflex femoral artery,method and their clinical application that multiple soft tissue defects of hand are repaire by muliplefoliated tissue flap only branch lateral circumflex femoral artery.MethodsFifteen patients with multiple soft tissue defects of hand were repaired muliplefoliated tissue flap only pedicled bydescending branch lateral circumflex femoral artery.At first,the anterolateral thigh perforator flap was designed and harvested according to the soft tissue defects of hand, then the descending branch lateral circumflex femoral artery was dissected at the same time the segmented perforator flap,fascia lata flap,rectus femoris muscle flap, vastus lateralis muscle flap, vastus intermedius muscle flap and distal spatium intermusculare flap were harvested in need according to distance among soft tissue defects.The muliplefoliated tissue flap was harvested only pedicled by descending branch lateral circumflex femoral artery, at last muscle flaps and fascia lata flaps were covered by skin graft, so the multiple soft tissue defects of hand were repaired in one time.ResultsNo vascular crisis happened. All skin grafts survived well, the contour of all repaired soft tissue defects was good and protective feeling was recovered by skin grafts of all flaps. All cases were got follow-up and the range was from 6 to 20 months(the average was 8.7 months).Wound of donor site healed well, muscle strength of quadriceps and motion of knee were normal. Three cases were excellent,nine cases were well and 3 cases were good, according to upper extremity function evaluation criteria of Chinese Medical Society for the Surgery of the Hand, the rate of good was 80 percent.ConclusionMultiple soft tissue defects of hand can be repaired by muliplefoliated flap only pedicled by descending branch of lateral circumflex femoral artery. Its advantages included reduction of operation time and treatment, good recovery of hand contour and function. It is a good method to repair multiple soft tissue defects of hand.

Objective To find out the current thyroid volume of school children and its influencing factors in Chongqing.Methods Probability sampling method(PPS) was used to select 30 counties in Chongqing in 2011.Forty children aged 8-10of 1 randomly selected school from every county chosen were enrolled in the study.Thyroid volume of children was examined by B-ultrasonograghy.Body height and body weight were measured.The relationship between gender,age,height and weight and thyroid volume was analyzed,respectively.Results One thousand three hundred and twenty-two children aged 8-10 were investigated.The median of thyroid volume was 3.53 ml.The goiter rate was 5.52％ (73/1322).Thyroid volume of female and male was 3.55 and 3.51 ml,respectively.There was no significant difference of thyroid volume between female and male (H =0.68,P ＞ 0.05).Thyroid volume of children aged 8,9 and 10 was 3.30,3.53 and 3.76 ml,respectively.There was a significant difference of thyroid volume among different age groups(H =52.49,P ＜ 0.01).Thyroid volume of children height (110-,120-,130-and ≥140 cm,respectively) was 2.96,3.22,3.59 and 4.13 ml.There was a significant difference of thyroid volume among different height groups (H =149.23,P ＜ 0.01).Thyroid volume of children weight(17-,20-,30-and ≥40 kg,respectively) was 2.71,3.31,3.91 and 4.74 ml.There was a significant difference of thyroid volume among different weight groups(H =138.44,P ＜ 0.01).For the coefficients of simple and partial correlation,there was a significant correlation between thyroid volume and age,height and weight (P ＜ 0.05).The Spearman coefficient was 0.2411,0.3950 and 0.4285,respectively.The partial correlation coefficient was 0.0640,0.1154 and 0.2319,respectively.The standard partial coefficient of age,height and weight was 0.640,0.1154 and 0.3410,respectively.The proportion of the standard partial coefficients was 1 ∶ 1.8 ∶ 5.3.The function of body weight to thyroid volume was 5.3 times that of age and 3.0 times that of body height.Conclusions The goiter rate of schoolchildren in Chongqing is relatively high.Thyroid volume is affected by age,body height and body weight.The relationship between thyroid volume and iodine nutrition needs further study.

Objective: To observe the effects of acupoint thread-embedding therapy and low-carbohydrate diet therapy on obese patients with food addiction. Methods: Sixty-five eligible patients were randomized into a thread-embedding group of 33 cases and a diet group of 32 cases to respectively receive 12-week treatment. Before treatment, after treatment and at 6-month follow-up, the two groups were observed and compared in terms of body mass (BM), waist circumference (WC), hip circumference (HC), waist-to-hip ratio (WHR), body mass index (BMI), body fat rate (BFR), basal metabolic rate (BMR) and Yale food addiction scale version 2.0 (YFAS 2.0). Results: At the end of treatment, there were no significant differences in the general efficacy, and the improvements in BM, BMI, WC, HC, WHR and BFR between the thread-embedding group and diet group (all P>0.05). At follow-up, the thread-embedding group showed more significant improvements in all the aforementioned indicators compared with the diet group except HC (all P<0.05). At the end of treatment and follow-up, BMR and YFSA 2.0 had more significant improvements in the thread-embedding group than in the diet group (all P<0.05). Conclusion: Acupoint thread-embedding therapy can produce significant efficacy in treating obese patients with food addiction; it can improve the food addiction state and work better in maintaining the efficacy compared with low-carbohydrate diet therapy.

BACKGROUNDBrachial-ankle pulse wave velocity (baPWV) is a reliable method for measuring arterial elasticity, but the absence of reference value for baPWV has limited its wide use. We conducted an epidemical study in north China to investigate the reference value of baPWV for Chinese people and its influential factors.

METHODSA total of 974 identified healthy subjects were recruited in this study. The values of baPWV were evaluated noninvasively with an automatic device.

RESULTSFor healthy population, the mean value of baPWV was higher for male (P < 0.001). Multiple regression analysis demonstrated that both age and systolic blood pressure were positively associated with baPWV for male and female (P < 0.001). BaPWV value was higher in male than in female in younger group (< 50 years) but not in older group (P

CONCLUSIONSAging is the most important reason of arterial stiffness, but the effect of age on baPWV augmentation is greater for healthy female than their male counterpart. The reference values of baPWV by sex and age are very useful for clinical and preventive medicine.

Adult ; Age Factors ; Aged ; Ankle Brachial Index ; Asian Continental Ancestry Group ; Female ; Humans ; Male ; Middle Aged ; Reference Values ; Regression Analysis ; Sex Factors

OBJECTIVETo study the effects of dexamethasone on intracellular expression of Th17 cytokine interleukin 17 and the mechanisms in asthmatic mice.

METHODSExperimental asthma was induced by ovalbumin (OVA) sensitization in 20 in female Balb/c mice with (dexamethasone group, n=10) or without dexamethasone treatment (model group, n=10), with another 10 serving as the control group. The levels of IL-17 in the bronchoalveolar lavage fluid (BALF) and serum of the mice were measured by enzyme-linked immunosorbent assay (ELISA), and the airway inflammation was evaluated by HE staining. The expressions of IL-17 and RORgammat mRNA were measured by reverse transcription-polymerase chain reaction (RT-PCR), and the expression of RORgammat protein was measured by immunohistochemical staining.

RESULTSThe levels of RORgammat and IL-17 mRNA and protein in the asthmatic model group were significantly higher than those in the control group (P<0.01), and the increased expressions of RORgammat and IL-17 mRNA and protein in the asthmatic mice were significantly reduced by dexamethasone treatment (P<0.05).

CONCLUSIONDexamethasone can inhibit the release of IL-17 probably by inhibiting RORgammat expression and blocking Th17 differentiation in asthmatic mice.

Animals ; Asthma ; chemically induced ; immunology ; metabolism ; Dexamethasone ; pharmacology ; Female ; Interleukin-17 ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; metabolism ; Ovalbumin ; RNA, Messenger ; biosynthesis ; genetics ; T-Lymphocyte Subsets ; immunology ; metabolism ; T-Lymphocytes, Helper-Inducer ; immunology ; metabolism

BACKGROUND: Studies have shown that dental pulp stem cells have high proliferation and multi-directional differentiation abilities and can differentiate into a variety of cells under certain conditions. At present, the use of dental pulp stem cells to construct tissue-engineered dentin-pulp complex is expected to become a new strategy for human dental defect repair . OBJECTIVE: To observe the effect of dental pulp stem cells on the repair of rat tooth defects by construction of tissue-engineered dentin-pulp complex. METHODS: Twenty-four Sprague-Dawley rats were used to make animal models with dental pulp removal, and then model rats were randomly divided into model group and transplantation group. Rats in the transplantation group were subjected to tissue-engineered dentin-pulp complex transplantation, and those in the model group given no treatment. Tooth samples were collected at 3, 5, 7 weeks post transplantation and observed using hematoxylin-eosin staining and immunofluorescence staining. The dentin thickness of rats was measured by Image Pro Plus 6.0 image software system. RESULTS AND CONCLUSION: (1) Dental pulp cells was mostly spindle/oval-shaped and partially polygonal. The third generation of cells with long spindle shape showed fibrous growth and uniform morphology. Findings from immunohistochemical staining showed spindle-shaped deep-colored cells with oval nuclei stained as dark blue were identified as fibroblast-like cells, and were positire for vimtin. (2) Findings from hematoxylin-eosin staining showed vacuolar degeneration of the cells, and hbdestroyed pulp tissue and debris, irregular cord-like tissue, and a large amount of red blood cells and inflammatory cells in the pulp cavity, accompanied by clearly visible vascular dilation. Seven weeks after transplantation, a bundle of odontoblasts were visible in the matrix-like tissues of the dentin, and there was a distinct boundary between the original dentin and regenerated dentin. (3) Findings from immunofluorescent staining showed that after dentin-pulp complex transplantation, the number of cells in the pulp cavity increased significantly at 3 weeks, and there was also a substantial increase in dental pulp cells at 5 weeks that were distributed on the wall of the pulp cavity. Compared with the model group, the dentin thickness in the transplantation group was significantly higher at each time after transplantation (P < 0.05), and in the transplantation group, there was also a significant difference in the dentin thickness at different time points (P < 0.05). To conclude, the tissue-engineered dentin-pulp complex can promote dentin regeneration and repair.