Objective To eonstruct three eukaryotic expression vectors containing wilde-type hGHR gene and its mutants(hGHR-E42K and hGHR-H56R) related to congenital growth hormone insensitivity, then check their expression in CHO cells. Methods With the available PUC-hGHR vector, two mutate hGHR genes (hGHR-E42K and hGHR-H56R) were obtained through mutagenesis. Then three recombinants were cloned into eukaryotic expression vectors pcDNA3.1/zeo(+) with restriction enzymatic reactions.Then with Lipofectamine2000, we trans-fected expression vectors to CHO cells and screened the stably expressed CHO cells by Zeocin. RT-PCR and/or Western blotting were used to examine hGHR and STAT5-P. Results After sequencing, two mutations were introduced to hGHR, three eukaryotic expression vectors were identified. The transfected CHO cells expressed vd-hGHR or its mutants. Compared with hGH-wt, two mutate cells (E42K and H56R) had decreased phosphorylated STAT5 levels. Conclusion Three CHO cells which stably expresses wide type hGHR and its mutants were successfully established, E42K and H56R partly interfered the phosphorylation of STAT5.

Objective To evaluate the effect of abducens orthosis combined with walker on de-velopmental dysplasia of the hip ( DDH ). Methods A total of 126 patients (224 hips ) with DDH aged 6～36 months in Xiangya Hospital was randomly divided into 2 groups: an orthosis combined with walker group and an improved hip frog cast fixation group. Seventy patients (130 hips) were treated by the orthosis combined with walker and 56 patients (94 hips) were treated by the improved hip frog cast fixation. We compared the effect and complications of the 2 groups. Results The fine-ness rates of the orthosis combined with walker group and the improved hip frog cast fixation group were 89.2% and 90.4% , respectively, with no significant difference (P>0.05). The rate of femoral head osteonecrosis in the orthosis combined with walker group was significantly lower than that in the improved hip frog cast fixation group (1.5 % vs. 5.3 % , P<0.05) , but the re-dislocation rate in the former was significantly higher than that in the latter (6.9 % vs. 1.1% , P<0.05). Conclusion Both methods are effective for DDH. Orthosis combined with walker has a lower propor-tion of femoral head osteonecrosis, but a higher proportion of re-dislocation.

OBJECTIVE: To evaluate the relative factors influencing olfactory dysfunction in patients with chronic rhinosinusitis (CRS). METHOD: Visual analogue scale (VAS) was applied to measure the severity of olfactory dysfunction of 270 patients with CRS. Patients were divided into two groups, one was that the quality of life (QOL) of patients was affected by olfactory dysfunction (VAS > 5), the other was that without QOL affected by olfactory dysfunction (VAS ≤ 5). The association between age, gender, nasal polyps, allergic rhinitis, smoking history, early nasal surgery history and other clinical factors, and serum total IgE level, peripheral blood eosinophil count, peripheral blood mononuclear cell count and olfactory dysfunction was analyzed. RESULT: The number of patients with nasal polyps, allergic rhinitis, previous nasal surgeries, the level of serum total IgE, and the severity of edema were significantly increased in patients with impaired QOL associated with olfactory dysfunction (P < 0.05). Sex distribution, age, smoking history, deviation of nasal septum, eosinophil and mononuclear cell count did no statistically differ between the groups with and without impaired QOL associated with olfactory dysfunctions (P > 0.05). Serum total IgE increased (OR = 1.003, P < 0.01) and severe edema (OR = 2.483, P < 0.01) were the risk factors for the impairment of olfactory function, more notably for edema; whereas previous nasal surgeries was a protective factor (OR = 0.408, P < 0.01). CONCLUSION Sever edema and increased serum total IgE are risk factors, whereas previous nasal surgeries history is a protective factor for the olfactory dysfunction.
Chronic Disease ; Female ; Humans ; Leukocytes, Mononuclear ; Male ; Nasal Polyps ; Olfaction Disorders ; etiology ; Quality of Life ; Rhinitis ; complications ; immunology ; Rhinitis, Allergic ; complications ; immunology ; Risk Factors ; Sinusitis ; complications ; immunology ; Smell ; Smoking ; adverse effects

Objective To observe the effects of laparoscopy combined with choledochoscopy in the treatment of polypoid lesions of gallbladder and the effects on postoperative rehabilitation of patients. Methods 78 cases of patients with polypoid lesions of gallbladder treated in our hospital from August 2011 to November 2014 were selected as the study object. According to the order of medical treatment and hospital admission , the patients were numbered and randomly divided into laparoscopy group and combined group , with 39 cases in each group. The laparoscopy group were treated by laparoscopic cholecystectomy while the combined group were treated by laparoscopy combined with choledochoscopy gallbladder-protection surgery. The operation indicators , recovery of gastrointestinal function and incidence of postoperative complications in the two groups were observed. After discharge , the 2 groups of patients were followed up for 1 year. The improvement of postoperative digestive tract symptoms and rehabilitation of patients was observed. Results The intraoperative bleeding volume, postoperative fasting time, defecation time and hospitalization time of the combined group [(20.13 ± 7.48) mL, (0.51 ± 0.98) d, (1.20 ± 0.84) d, (1.98 ± 1.54)d] were less than those of the laparoscopy group (P <0.05); The incidence rate of complications in the combined group (10.26%) was lower than that in the laparoscopy group (33.33%) (P < 0.05); Immediately at the end of operation and 1 week after operation, VAS scores of the combined group [(4.23 ± 0.41), (2.46 ± 0.17) ] were lower than those in the laparoscopy group (P < 0.05); At the last follow-up, KPS score of the combined group increased to (89.98 ± 1.24), which was higher than that in the laparoscopy group (P < 0.05); GradeⅠpostoperative recovery in the combined group accounted for 87.18% which was higher than that in the laparoscopy group (64.10%). There was no case of grade III and the proportion was lower than that in the laparoscopy group (15.38%). The differences in proportions of grade I and III between the two groups were statistically significant (P < 0.05). Conclusion To adopt laparoscopy combined with choledochoscopy in the treatment of polypoid lesions of gallbladder , the curative effect is certain and the minimally invasive advantage is obvious. The long-term recovery effect is good and the safety is high.

A gas chromatography mass spectrometry (GC-MS) method has been developed and fully validated for the simultaneous determination of natural borneol (NB) and its metabolite, camphor, in rat plasma. Following a single liquid-liquid extraction, the analytes were separated using an HP-5MS capillary column (0.25 mm ? 30 m ? 0.25μm) and analyzed by MS in the selected ion monitoring mode. Selected ion monitor (m/z) of borneol, camphor and internal standard was 95, 95 and 128, respectively. Linearity, accuracy, precision and extraction recovery of the analytes were all satisfactory. The method was successfully applied to pharmacokinetic studies of NB after oral administration to Wistar rats.

Objective To observe the effect of astragaloside IV (A) and SB203580 antagonize cadmium (Cd) toxicity on expression of associated protein and blood-testis barrier(BTB) in rats and to study the protective mechanism of A on it.Methods Totally 21 SD male rats were randomly divided into 7 groups, 3 rats per group:Cd [ intraperitoneally injected with 0.1%CdCl2,1mg/(kg?d)],Cd+A [at the above dose of CdCl2,at the same time with A,10mg/(kg?d)], and Cd +SB203580 [at the above dose of CdCl2,at the same time with SB203580,100μg/(kg?d)], each of the above groups was further divided into continuous five and ten days treatment groups .The control group was intraperitoneally injected with equal dosage of normal saline .The testes were studied by light , electron microscopy , immunohistochemistry and Western blotting .Results In the control group ,irregular and lightly stained nuclei of Sertoli cell ( Sc) in seminiferous tubules were observed by HE staining .A continuous electron density line of tight junction ( TJ) and normal ultrastructure of BTB were observed .After Cd treatment ,the vesicular formation in the Sc was observed .The ultrastructural damage of Sc and TJ was observed .Compared with the corresponding time point of Cd group ,these were weakened in morphology of testis and ultrastructure of TJ after Cd +A or Cd +SB203580 treatment .The positive products of zonula occludens-1 ( ZO-1 ) and claudin-11 were localized mainly in the base of the seminiferous tubule .After Cd treatment , the average absorbance (AA) of ZO-1 and Claudin-11 was decreased significantly compared with that of the control group (P<0.05).After Cd +A or Cd +SB203580 treatment,AA of ZO-1 and Claudin-11 were increased significantly compared with that of the Cd group(P<0.05),though lower than that of the control group .The result of Western blotting showed that phosphorylation-p38MAPK in Cd group was increased significantly compared with that of the control group (P<0.05).After Cd +A or Cd+SB203580 treatment, it was decreased significantly compared with that of the Cd group (P<0.05).Conclusion Cd decreases ZO-1 and Claudin-11 expression and damages ultrastructure of TJ in BTB , asⅣhas protective effect on it , and is related to inhibiting activation of p 38 MAPK pathway .

Objective To investigate the variation of chemical characteristics with environmental factors and establish a relationship betweem morphological characters and chemical composition of Prunella vulgaris collected in different areas of China.Methods Twelve phenotypic traits and three chemical compositions were assessed in 28 populations of P.vulgaris collected from different locations in China.Results The variability ranges observed at phenotypic and chemical levels were polymorphic.According to the morphological traits,28 populations of P.vulgaris could be grouped into six clusters,and two morpho-types could be clearly distinguished.Perceptible differences could be discerned in the plant height,leaf length,corolla length,calyx length,fruiting spikes length,and maturity period.Based on three kinds of components including ursolic acid,total flavonoids,and total polysaccharides,all populations could be identified as four types.Cluster Ⅳ showing high content of ursolic acid,total flavonoids,and total polysaccharides could be utilized to develop superior derivatives.Conclusion The variation of chemical characteristics is influenced by the genetic and environmental factors,such as soil,climate,longitude,and altitude.It provides a solid basis for efficiently evaluating qualities and establishing good agricultural practices for P.vulgaris.

Objective To construct three eukaryotic expression vectors containing wilde-type hGHR gene and its mutants(hGHR-E42K and hGHR-H56R) related to congenital growth hormone insensitivity,then check their expression in CHO cells. Methods With the available PUC-hGHR vector,two mutate hGHR genes (hGHR-E42K and hGHR-H56R) were obtained through mutagenesis. Then three recombinants were cloned into eukaryotic expression vectors pcDNA3.1/zeo(+) with restriction enzymatic reactions. Then with Lipofectamine2000,we transfected expression vectors to CHO cells and screened the stably expressed CHO cells by Zeocin. RT-PCR and/or Western blotting were used to examine hGHR and STAT5-P.Results After sequencing,two mutations were introduced to hGHR,three eukaryotic expression vectors were identified. The transfected CHO cells expressed wt-hGHR or its mutants. Compared with hGH-wt,two mutate cells (E42K and H56R) had decreased phosphorylated STAT5levels. Conclusion Three CHO cells which stably expresses wide type hGHR and its mutants were successfully established,E42K and H56R partly interfered the phosphorylation of STAT5.

Objective To investigate the neuronal nitric oxide synthase (nNOS) distribution in intramural striated sphincter of female rats. Methods 4 female Wistar rats were anesthetized with chloral hydrate. The whole bladder and urethra specimen were harvested, fixed in formalin and imbedded in paraffin wax. Longitudinal and transverse sections were developed with 2 urethra respectively. Routine HE staining and Mallory phosphotungstic acid-hematoxylin staining were used for observation of urethral microstructure. nNOS distribution in intramural striated sphincter was determined with immunohistochemical staining. Results The urethras of female Wistar rat were tubular structure about 1.5～2.0 cm long. From outside inward,the structure included circular striated muscle layer,circular smooth muscle layer,longitudinal smooth muscle layer,dense connective tissue and epithelium. The longitudinal sections showed intramural striated muscle predominates the middle and distal third,not the whole urethral length. The transverse sections showed intramural striated muscle is closed ring shape,about 5～10 layers uneven distributed with more layers in the posterior wall than in the front wall. Strong positive expression of nNOS immunoreactivity was found in some intramural striated muscle fiber. Conclusion nNOS expresses in some intramural striated muscle fiber, surggesting nitric oxide may play an important role on intramural striated sphincter.

BACKGROUND:Numerous studies have shown that air wave pressure therapy plays an important role in prevention of deep venous thrombosis of lower limb after a major operation on the hip.
OBJECTIVE:To explore the efficacy of air wave pressure therapy on deep vein thrombosis of the lower extremity in patients with total knee arthroplasty.
METHODS:A total of 80 patients with total knee arthroplasty were randomly divided into experimental group and control group. Patients in the control group were treated only with conventional method to help the patient massage the lower extremity by the nurse and combined with active functional exercise;patients in the experimental group were treated with air wave pressure therapy at 2 days postoperatively, and were also treated with conventional method to help the patient massage the lower extremity by the nurse and combined with active functional exercise. Swel ing index, coagulation index and the number of patients with deep venous thrombosis of the lower extremity were recorded in both groups.
RESULTS AND CONCLUSION:At 2 weeks after treatment, swel ing index, coagulation index and the number of patients with deep venous thrombosis of the lower extremity were significantly reduced in the experimental group, and its effects were significantly better than those in the control group (P<0.05). Early use of air wave pressure therapy after total knee arthroplasty can al eviate limb swel ing effectively and have obvious advantages in improving coagulation index and blood coagulation condition and in preventing deep vein thrombosis of the lower extremity after total knee arthroplasty.