The polar tube protein is the major component of polar tube, and can specifically locate on the polar tube of microsporidia and plays an important role in invasion host cell. In this study, we analyzed the potential O- and Nglycosylation sites in polar tube protein 1 from Nosema bombycis. NbPTP1 was successfully cloned to eukaryotic expression vector pMT/Bip/V5-His A, involved V5 and His tags. After transfection, NbPTP1 gene could be efficiently expressed in Drosophila S2 cells. In addition, Lectin blotting and beta elimination analysis showed that NbPTP1 expressed in Drosophila S2 cells was O-glycosylation. These studies provided a basis for understanding the relationship between glycosylation and function of NbPTP1, helped us to reveal the infection mechanism of microsporidia and established effective diagnosis and prevention methods for microsporidia.

Objective To develop a new method to determine the contents of rutaecarpine in Fuzhengpingxiao capsule by HPLC method.Methods Samples were handled by ethanol and extraction with ethyl acetate.The separation was achieved on an Agilent TC-C18column using a mobile phase system of acetonitrile-water(2% Tetrahydrofuran and 0.2 % formic acid)at a flow rate of 1.0 ml/min.The temperature of column was 40 ℃ and the detection wavelength was 240 nm.Results The cali-bration curves of rutaecarpine showed good linearity in the ranges of 1.18-118 μg/ml,r=0.999 9.The results of intra-day and inter-day precisions were both within 2%,the average additional recovery rate was 94.20%.Conclusion The HPLC method was accurate,specific,sensitive and reproducible,which could be used for quality control of rutaecarpine in the preparation of Fuzhengpingxiao capsule.

Objective To explore method for determination of calycosin contents in Wujin capsule by HPLC .Methods The sample was studied on ZORBA Bonus‐RP (4 .6 mm × 260 mm ,5 μm) column ,the mobile phase consisted of acetonitrile and H2O‐0.1% HCOOH (gradient elution) ,the temperature of column was 25 ℃ ,the UV detection wavelength was set at 260 nm , flow speed was 1 .0 ml/min and injection volume was 10 μl .Results The calycosin calibration curve was at the range of 7 .4‐117 .6 μg/ml (r=1 .000) ,with their regression function being Y=34 .002 X-3 .451 2 .The recovery of calycosin was 95 .24%(RSD=2 .10% ) .6 batches of Wujin capsule calycosin contents were 9 .26‐23 .14μg/g .Conclusion HPLC was an accuracy and simple method for determining the content of calycosin in Wujin capsule .

Objective To establish a method for the determination of asiatic acid in the root of Actinidia valvata . Methods HPLC-VWD was used in the analysis .The column was Agilent HC-C18 (4 .6 mm × 250 mm ,5μm);the mobile phase was acetonitrile :(30 mmol/L) acetic acid amine solution (35:65);the flow rate was 1 .0 ml/min;the temperature of column was 25 ℃ ;the detection wavelength was set at 210 nm ;the injection volume was 25 μl;the running time was 35 min .Results
Asiatic acid was separated with interference in baseline .The linear range was 25 .30-506 .0 μg/ml with linear correlation of 0 .999 6 for asiatic acid .The result of intra-day and inter-day precisions were both within 5% (n=3) ,and the average recovery was 99.4% with RSD 1 .9% (n=6) .Conclusion The method was simple ,rapid ,accurate and convenient for quality control of asiatic acid in the root of Actinidia valvata .

Objective To study the distance and pattern of the movement of the maxillary teeth after 6 months driftodontics for dental crowding.Methods 20 dental crowding children,according to the treatment plan,who were required to removal four first premolars,were involved in this study (8 males,12 females,aged 11-14 years old).All of the children did not wear appliance in six months after teeth extraction.X-ray image data before and six months after teeth extraction were collected.The width of extraction space,the anterior irregular index and inclination of the axis of each tooth were recorded before and six months after tooth extraction.The data were analyzed by t-test and Pearson relative analysis.Results The study model showed that bilateral maxillary space of extraction pre-and pro-driftodontics decreased (5.79 ± 4.9) mm (P ＜ 0.01) ; anterior crowding decreased (5.79± 4.12) mm,irregular index was significantly reduced (P＜0.01).Cephalometric film measurement showed that canine crown moved distally (4.87±3.54) mm after the drift (P＜0.01).Canine tooth axis changed from mesial to upright.The crown of maxillary first molar moved mesially (1.18± 0.52) mm (P ＜ 0.01).There were no significant changes in apical position.Conclusions After 6 months of the physiological drift,upright moving of canine relieves anterior teeth crowding.The molar anchorage is saved.

A 31-year-old male normotensive patient with aldosterone-producmg adenoma complained of thirst,polydipsia,polyuria,and periodical paraplegia.The diagnosis is raised by signs of hypokalemia.Despite the lack of hypertension,primary aldosteronism was confirmed by persistent hypokalemia,increased urinary potassium,increased urinary and plasma aldosterone levels and suppressed plasma rennin activity (PRA).The blood pressure profile was studied by ambulatory monitoring,and the mean blood pressure of 24h was normal and the circadian rhythm remained normal. Surgical removal of the histologically typical aldosterone-producing adenomas normalized the kalemia.The patient had a marked fall in blood pressure with mean values of 21/17 mm Hg ( diurnal and nocturnal blood pressure were 19/17 and 22/17 mm Hg respectively)and recovery of normal urinary and plasma aldosterone levels and PRA 6 weeks after surgery.This suggests that excess serum aldosterone induced relative hypertension in those patients whose blood pressure was spontaneously very low.Our observations call for primary hyperaldosteronism assay in patients with hypokalemia and renal potassium leakage.

Objective To analyze the clinical characteristics and prognosis of patients with insulin autoimmune syndrome (IAS) and to get better understanding of IAS by literatures reviewing.Methods Nine cases of IAS who were diagnosed in the General Hospital of the People's Liberation Army from 2001 to 2011 were analyzed.Results All patients had hypoglycemic syndrome and episodes of hypoglycemia during both postprandial and fasting states.Most cases (8/9) were accompanied with autoimmune diseases,including 6 cases of Graves' disease treated with methimazole.The biochemical data showed extremely elevated serum insulin level (9/9) during hypoglycemic episode.For most patients,the tests of insulin autoantibodies were positive (7/9) while results of imaging examinations were negative(8/9).After removal of possible offending medications and with diet treatment,hypoglycemic episodes were ameliorated in 5 of 9 cases.For severe patients,acarbose (1/9) and prednisone (3/9)therapy were useful.During the period of follow-up,four cases experienced no episode of hypoglycemia and 3 cases with markedly reduced episodes.Conclusions IAS is characterized by hypoglycemic episodes,elevated blood insulin levels,and positive insulin autoantibodies.It is strongly related with autoimmune disease and is able to be induced by methimazole.Most patients undergo remission after diet treatment,drug withdrawal,and oral prednisone.

Objective To study gender-related differences of clinical characteristics and vascular complications in patients with aldosterone-producing adenoma(APA).Method Consecutive 315 patients with APA confirmed by pathological diagnosis were included and the clinical features and vascular complications were compared based on gender.Results (1) Of the 315 patients with APA,female accounted for 52.7％ (166/315).Male patients with APA presented a higher BMI,compared with females [(25.4± 2.9) vs (24.1 ± 3.2) kg/m2,P＜0.01],and the history of smoking and drinking was more common in male patients.No significant difference was found in regard to the age,duration,hypokalemia,tumor size,and family history of hypertension between two groups (P＞0.05).(2) No significant difference was found in the prevalence of hypertension,duration of hypertension,blood pressure,and antihypertensive medieation between two groups.However,the patients with grade 3 hypertension tended to be more prevalent in males(P =0.08).(3) Serum aldosterone concentrations were similar in two groups,but 24 h urinary aldosterone showed an increased trend in male group (P =0.07).(4) The overall prevalences of cardiovascular (51.0％ vs 36.1％) and cerebrovascular (9.4％ vs 3.0％) complications were significantly higher in male group (P ＜ 0.05).Further analysis of cardiac events revealed significantly higher rates of left ventricular hypertrophy (23.5％ vs 13.9％) and arrhythmia (21.5％ vs 10.8％) in males (P＜0.05).However,no significant difference in the prevalence between two groups was found in regard to cerebral hemorrhage,infarction,and chronic renal insufficiency.(5) The fall of blood pressure and recovery from hypokalemia were comparable between males and females in two weeks after sugery.However,in the patients with persistent hypertension,the number of antihypertensive drugs used in males was greater than that in females and more male patients needed ≥ 2 types of antihypertensive drugs.Conclusion There were significant gender-related differences with regard to clinical features and vascular complications in patients with APA.

An offline two-dimensional system combining a rat cardiac muscle cell membrane chromatography time-of-flight mass spectrometry (CMC-TOF/MS) with a high performance liquid chromatography time-of-flight mass spectrometry (HPLC-TOF/MS) was established for investigating the parent components and metabolites in rat urine samples after administration of the roots of Aconitum carmichaeli.On the basis of the analysis of the first dimension,retention components of the mine sample were collected into 30 fractions (one fraction per minute).Then offline analysis of the second dimension was carried out.34 compounds including 24 parent alkaloids and 10 potential metabolites were identified from the dosed rat urine,and then binding affinities of different compounds on cell membranes were compared and influences of some functional groups on activity were estimated with the semi-quantification and curve fitting method.As a result,binding affinities decreased along with the process of deacylation,debenzoylation and demethylation,which may be related to the alleviation of toxicity in the procedure of herb processing or metabolism.Moreover,some minor components in rat urine (Songorine,14-benzoylneoline,Deoxyaconitine,etc.) exerted relatively strong affinity on cell membranes are worth exploring.The results delivered by the system suggest that the CMC can be applied to in vivo study.

Objective To investigate the gender-related differences in patients with Graves'disease (GD).Methods Seven hundred and eighty-seven consecutive patients with GD treated with anti-thyroid drug in out-patient clinic of department of endocrinology of Chinese PLA general hospital from Oct.2009 to May 2010 were included in the study.The diagnosis of GD WSB made according to common clinical and laboratory criteria.Results (1)In Graves'disease,a marked female preponderance Was found.The female/male ratio was 3.35:1.Female patients with GD presented at a younger age,compared with males[(33.20±13.07 vs 36.75±13.89)year,P=0.002],and the peak incidences of the disease occurred in female and male were 20-29 years and 40-49 years of age respectively.(2)Male patients presented a higher prevalence of weight loss,thyrotoxic cardiopathy,periodic paralysis,and non-invasive ophthalmopathy whereas tachycardia was more common in females.(3)The females presented larger diffuse goiters with higher serum TT4,FT4,and thyrotropin receptor antibody(TRAb)levels.Cases with TRAb>30 U/L were more common in females,whereas those with TRAb 10-30 U/L were frequently seen in males.(4)There was no gender-related difference in the family history of GD.(5)Remission atleast 6 months after anti-thyroid drug therapy in 211 patients,there was a relapse rate of 64.3%(137/211)without gender difference.Conclusion GD appeared at a younger age and with larger diffuse goiters in females,together with more marked elevation of serum TT4,FT4 and TRAb levels.Altogether,these findings depict different patterns for GD in males and females.