OBJECTIVE: To establish a HPLC method for the determination of content of entecavir and its related substances. METHODS: The HPLC condition was consisted of a Luna C18 column with a column temperature at 25 ℃, using gradient eluate method, the mobile phase was acetonitrile,with a flow rate of 1.0 mL?min-1 and detection at 254 nm. RESULTS: The calibrated linear curve of entecavir was within 0.033 45～0.167 2 mg?mL-1(r=0.999 98). The average recovery was 100.09%,RSD=0.20%, The content of related substance was 0.54%～1.48%. CONCLUSION: This accurate and reliable HPLC method is applicable for the quality control of entecavir and its related substances.

Objective To explore the correlation between serum uric acid levels and uric acid excretion indexes in children with primary nephrot?ic syndrome(PNS),and to reveal the prevalence of hyperuricemia and its possible causes in PNS. Methods The clinical data in 74 cases of pe?diatric PNS were retrospectively analyzed. These patients were grouped as hyperuric acid(HUA)or non?hyperuric acid(NUA),and their clinical features and laboratory findings were compared between two groups,including age,serum uric acid,serum creatinine,urea,cystatin C,triglycer?ide,cholesterol,albumin,creatinine clearance rate,24 hours urine uric acid,serum uric acid/serum creatinine,uric acid clearance,and the frac?tional excretion of uric acid and other indicators of differences. In addition ,a correlation analysis was carried out for the excretion of uric acid index. Results Several factors were lower in HUA group than that of NUA group ,such as the serum albumin and fractional excretion of uric acid(P1=0.034,P2=0.025);while the serum cystatin C and serum uric acid/serum creatinine ratio(P1=0.038,P2=0.001)was higher in HUA group. The uric acid excretion was negatively correlated with serum uric acid levels in all 74 children with PNS(r=-0.43,P=0.016),and the serum uric acid/creatinine ratio was positively correlated with serum uric acid(r=0.486,P=0.001). Conclusion Both the increase of blood uric acid production and excretion reduction were observed in children with PNS ,and the serum uric acid levels and renal tubular function is closely related.

OBJECTIVETo observe the difference in acupuncture for pain threshold at different time points among the groups of 9 TCM constitutions.

METHODSThe cross-sectional survey was adopted to investigate TCM constitutions among 600 subjects and determine 9 TCM constitution types (neutral constitution, qi-deficiency constitution, yang-deficiency constitution, yin-deficiency constitution, phlegm-damp constitution, damp-heat constitution, blood-stagnation constitution, qi-stagnation constitution, special diathesis constitution). The same acupuncture manipulation was applied to Zusanli (ST 36) on the left side in the subjects and the needle was retained for 30 min. The tenderness threshold was detected with 2390 type Von Frey apparatus at different time points, named before acupuncture, at the moment after qi arrival, in 10 min of needle retaining, in 30 min of needle retaining and in 15 min after needle withdrawal in the subjects of 9 TCM constitutions.

RESULTSThe interactive effect happened between the constitution type and time point (P < 0.05). Among the groups of 9 TCM constitutions, the pain threshold values at the moment after qi arrival (except blood-stagnation constitution, qi-stagnation constitution, special diathesis constitution) in 10 min of needle retaining and in 30 min of needle retaining were increased as compared with those before acupuncture separately (P < 0.01), among which, the value increase was the most significant in 30 min of needle retaining. The differences in the pain thresholds were significant in 15 min after needle withdrawal in the groups of neutral constitution and damp-heat constitution as compared with those before acupuncture (both P < 0.01). In 10 min of needle retaining and in 30 min of needle retaining, as compared with the group of neutral constitution, the changes in pain thresholds of the rest abnormal constitutions were apparently lower (all P < 0.05).

CONCLUSIONAcupuncture at Zusanli (ST 36) presents different effects among the groups of different constitution types. The effect maintaining durations are different.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Cross-Sectional Studies ; Female ; Humans ; Male ; Pain Management ; Pain Threshold ; Yang Deficiency ; therapy ; Yin Deficiency ; therapy ; Young Adult

Objective To determine the extracellular enzymatic activity of common pathogens for onychomycosis, in the hope of finding virulence factors associated with the pathogenesis of onychomycosis. Methods Strains tested in this study included standard strains of common dermatophyte and non-dermatophyte fungi as well as clinical isolates of Trichophyton rubrum from patients with onychomycosis. All the tested strains were cultured in medium containing nail fragments at 25 ℃ for 10 to 21 days followed by the determination of the nail fragment-containing medium, a significant increase was observed in the activities of esterase, esterase lipase, N-acetyl-β-glucosaminidase and α-mannosidase in dermatophytes compared with non-dermatophytes (all P ＜ 0.05 ), as well as in the activity of N-acetyl-β-glucosaminidase in Trichophyton rubrum compared with the other tested species of fungi (all P ＜ 0.05). No significant difference was noted in the activity of extracellular enzymes, except for that of naphthol-AS-BI-phosphohydrolase, between the isolates of Trichophyton rubrum from patients with different ranges of scoring clinical index for onychomycosis (SCIO). Conclusions In specific conditions, the extracellular enzymatic activity of fungi isolated from patients with onychomycosis is associated with fungal species, and may have a certain influence on the manifestations of anychomycosis.

Objective To observe the effects of deep electroacupuncture on carlilage tissue in knee osteoarthritis (KOA) rabbits. Meth-ods 40 New Zealand rabbits were randomly divided into normal group (A, n=10) and model group (n=30). The model group was modeled KOA with Hulth-Telhag way, and identified with X-ray. Then they were divided into no-treated group (B, n=10), deep electroacupuncture group (C, n=10) and routine electroacupuncture group (D, n=10) randomly. The groups C and D accepted electroacupuncture since 6 weeks after modeling, for 4 weeks. They were measured with pH of joint fluid, observed structure and pathology of cartilage under transmission electron microscope, detected apoptosis index, and determined the expression of acid-sensing ion channel 1 (ASIC1), p38 mitogen-activated protein kinases (p38MAPK) and p53 with Western blotting, and distribution of ASIC1 with immunohistochemistry in cartilage tissue. Re-sults The pHs of joint fluid from high to low were ranged as the groups A=C>D>B (P<0.01). The cartilage structure was more complete in the groups A, C and D than in the group B. The apoptosis rates from less to more were ranged as the groups A=C

In China.traditional Chinese medicines (TCMs) have been used in clinical applications for thousands of years.The successful hyphenation of high-performance liquid chromatography (HPLC) and mass spectrometry (MS) has been applied widely in TCMs and biological samples analysis.Undoubtedly.HPLC/MS technique has facilitated the understanding of the treatment mechanism of TCMs.We reviewed more than 350 published papers within the last 5 years on HPLC/MS in the analysis of TCMs.The present review focused on the applications of HPLC/MS in the component analysis,metabolites analysis,and pharmacokinetics of TCMs etc.50% of the literature is related to the component analysis of TCMs,which show that this field is the most popular type of research.In the metabolites analysis,HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry has been demonstrated to be the powerful tool for the characterization of structural features and fragmentation behavior patterns.This paper presented a brief overview of the applications of HPLC/MS in the analysis of TCMs.HPLC/MS in the fingerprint analysis is reviewed elsewhere.

This study evaluated the change in regional left ventricular myocardial function in rats following acute occlusion of the left anterior descending coronary artery (LAD) by using two-dimensional speckle tracking imaging (2D-STI). Sixty Wistar rats were randomly divided into two groups, a myocardial infarction (MI) group, in which 50 rats were subjected to LAD occlusion for 30-45 min, and a sham-operated (SHAM) group that contained 10 rats serving as control. Echocardiography was performed at baseline and 1, 4 and 8 week(s) after the operation. High frequency two-dimensional images of left ventricular short axis at papillary muscle level were recorded. Peak systolic radial strain (PRS) and circumferential strain (PCS) were measured in the mid-ventricle in short-axis view by using EchoPAC workstation. Left ventricular internal diameter at diastole (LVIDd) and systole (LVIDs), fractional shortening (FS), ejection fraction (EF) and left ventricular mass (LVM) were measured by anatomical M-model echocardiography. Infarct size was measured using triphenyl tetrazolium chloride (TTC) staining 1 week and 8 weeks after the operation. Fibrosis of left ventricular myocardium was displayed using Van Gieson staining 1 week after the infarction. In terms of the TTC staining results, the left ventricle fell into three categories: infarcted, peri-infarcted and remote myocardial regions. Compared with those at baseline and in the SHAM group, (1) PRS and PCS in the infarcted, peri-infarcted and remote myocardial regions were significantly decreased in the MI group within 1 week after the operation (P<0.05) and the low levels lasted 8 weeks; (2) Compared with those at baseline, LVIDd, LVIDs, FS, EF and LVM in the MI group showed no significant difference 1 week after the operation (P>0.05). However, LVIDd, LVIDs and LVM were increased significantly 4 and 8 weeks after the operation (P<0.05), and FS and EF were decreased substantially (P<0.05). Van Gieson staining showed that fibrosis developed in all the three myocardial regions to varying degrees. It is concluded that 2D-STI is non-invasive and can be used to assess regional function of myocardium with different blood supply in rats following acute occlusion of the LAD, and can be used as a sensitive and reliable means to follow up the process of left ventricular remodeling.

Objective To investigate the substantia nigra (SN) and brainstem raphe (BR) echogenic features of Parkinson's disease (PD) patients with musculoskeletal pain.Methods A total of 115 PD patients recruited in the Second Affiliated Hospital of Soochow University from October 2014 to May 2016 were assessed with the following rating scales:Unified Parkinson's Disease Rating Scale (UPDRS),Hoehn and Yahr Staging Scale (H/Y),Hamilton Rating Scale for Depression (HRSD),Beck Depression Inventory Ⅱ (BDI-Ⅱ) and Visual Analogue Scale (VAS).All the subjects underwent transcranial sonography during the clinical evaluation.And the patients were divided into PD with musculoskeletal pain (n =54) and PD without musculoskeletal pain (n =61) groups,or PD with depression(n =74) and PD without depression(n =41) groups.Results Compared with PD patients without pain,PD patients with musculoskeletal pain had higher scores of UPDRS-Ⅱ,-Ⅲ,HRSD,BDI,NMSQ and H/Y (UPDRS-Ⅱ score:12.56 ±6.01 vs 8.79 ±4.38,t =-3.801,P ＜0.01;UPDRS-Ⅲ score:24.43 ± 12.43 vs 20.07 ± 11.12,t=-1.986,P=0.049;HRSD score:11.65-±6.94 vs 8.38-±5.36,t=-2.844,P=0.005;BDI score:14.09 ±6.20 vs 9.74 ±6.00,t =-3.826,P ＜0.01;NMSQ score:8.57 ± 4.06 vs 5.60 ± 3.38,t=4.193,P＜0.01;H/Y:2.0(1.5,2.6) vs 1.5(1.0,2.0),Z=-3.011,P=0.003).Positive BR was more frequent in depressed than in non-depressed PD patients without pain (63.6％ vs 14.3％;x2 =15.25,P ＜0.01).Positive BR was positively associated with sex(r =0.228,P =0.014),age(r =0.184,P =0.049),disease duration (r =0.196,P =0.035),and depression (r =0.396,P ＜ 0.01).However,positive BR did not correlate with musculoskeletal pain.No correlation was found between positive SN and clinical characteristics of PD patients.Conclusions PD patients with musculoskeletal pain have worse activity of daily living,more severe motor symptoms,more non-motor symptoms,and are more depressed.SN and BR echogenecity do not correlate with musculoskeletal pain,however,hypoechogenic or interrupted BR is associated with depression in PD patients.

Objective To explore the value of two-dimensional strain echocardiography for quantitative assessing the change of regional left ventricular myocardial function in rats following acute myocardial infarction. Methods Sixty Wistar rats were randomly divided into two groups. The study group consisted of 50 rats with occlusion of LAD for 30-45 minutes and the sham-operated group consisted of 10 rats without occlusion of LAD. Echocardiography were performed before operation, which was defined as baseline, and 1 week, 4 weeks and 8 weeks after operation. Left ventricular internal diameter at diastole ( LVIDd) and systole < LVIDs), fractional shortening( FS), ejection fraction (EF) and left ventricular mass(LVM) were measured by anatomical M-model echocardiography. High frame rate two-dimensional images were recorded in the left ventricular short-axis views at the papillary muscle level. Peak systolic radial strain(PRS) and circumferential strain(PCS) of each segment were measured using 2-dimensional strain software. The rats were sacrificed and the infarcted size of each segment was measured using triphenyl tetrazolium chloride (TTC) after echocardiography was performed. Fibrosis of left ventricular myocardium was displayed using Van Gieson stain in 1 weeks after infarction. Results Based on the TTC findings,the left ventricle of the study group was divided into three regions:infarcted,peri-infarct and remote myocardial regions. Van Gieson stain showed fibrosis existed in all the three regions. Compared with baseline and sham-operated group, PRS and PCS of infarcted, peri-infarct and remote myocardial regions of the study group significantly decreased within 1 week after operation ( P <0. 01) and persisted for 8 weeks. PCS and PRS of infarcted, peri-infarct and remote myocardial regions of the study group in 4 weeks and 8 weeks after operation showed no significant difference when compared with those in 1 week after operation ( P >0. 01). Compared with baseline and sham-operated group,LVIDd,LVIDs and LVM of study group all increased significantly ( P <0. 05) in 4 weeks and 8 weeks after operation,and FS and EF reduced significantly ( P <0. 05). Two-dimensional strain obtained in interobserver and intraobserver both showed high agreement. Conclusions Two-dimensional strain echocardiography can assess regional function of myocardium with different perfusion in rats following acute myocardial infarction, and provides a sensitive and reliable method to follow up the process of left ventricular remodeling after myocardial infarction.

Objective To judge the effect of plasma exchange (PE) to the patients with severe hepatopath of nonage according to evaluating the change of the liver function of reserve with 13C-methacetin breath test. Methods There are two groups: the case group and the control group. Each group has 30 patients. The patients in the case group were treated by PE. All the patients received 13C-methacetin breath test at before or one week after treatment. MVmax40, CUM40 and CUM120 were present. At the same time, clinical symptoms, glutamate-pyruvate transaminase (ALT), total bilirubin (TBiL) and prethrombin active (PTA) were observed. Results MVmax40, CUM40, CUM120 and PTA were higher, ALT and TBiL were lower in the case group after treatment (t=4.679, 4.752, 5.048, 5.413, 6.208, 7.413, P=0.000,P<0.01). After a week, MVmax40, CUM40, CUM120 and PTA were higher, ALT and TBiL were lower in the case group than that in the control group (t=2.260, 2.247, 2.476, 4.017, 3.250, 3.658, P<0.05). The total effective rates in the case group and the control group were 83.3 % (25/30) and 43.3 % (13/30),which are significant different(χ2 10.335,P<0.01). Conclusion PE can im-prove the liver reservation functionin the severe hepatopath of nonage.