Objective To investigate the effect of ovariectomy and estrogen on secretion function and number of pancreatic islet beta cell in low-dose streptozotocin-induced diabetic rats. Methods Thirty female SD rats were randomly divided into five groups: normal control(NC) group, streptozotocin(STZ) group, ovariectomized(OVX) group, OVX + STZ(OS) group and OVX+STZ+estradiol(OSE) group. OVX, OS and OSE groups underwent ovariectomy, while NC and STZ groups underwent just sham operation. After surgery, OSE group was treated subcutaneously with estradiol 0.2 mg/kg twice weekly. At the end of 3 weeks, STZ, OS and OSE groups were induced by a single intraperitoneal injection of 40 mg/kg STZ. Then eight days later, plasma glucose and insulin levels were tested. The insulin protein, the average beta cell area and the relative beta cell mass were tested by streptavidin peroxidase conjugation method (SP). The quantification of beta cell apoptosis was performed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). The expressions of proliferating cell nuclear antigen(PCNA) protein, Bax and Bcl-2 were tested. Results With the administration of low-dose STZ, the plasma glucose was significantly higher and the insulin secretion curve after glucose loading, △I30/△G30 and modified beta cell function index(MBCI) were lower in OVX group than in other groups(all P<0.05). At the same time, the insulin protein, the relative beta cell mass and the beta cell area were dramatically decreased(all P<0.05). The beta cell apoptotic index was increased (t = 2.957, P< 0.05), the expression ratio of Bcl-2/Bax was decreased (0.41±0.03 vs. 0.76±0.05, P<0.05). Estrogen replacement therapy could obviously inhibit these changes. Compared with OS group, glucose disturbances and insulin secretion were improved dramatically in OSE group(all P<0.05); the insulin content, the relative beta cell mass and the average beta cell area were all enhanced (all P<0.05); the beta cell apoptotic rate was decreased(t =2.482, P<0.05), and the expression tatio of Bcl-2/Bax was increased (0.71±0.05 vs. 0.41±0.03, P<0.05). Conclusions The ovx rats are significantly more susceptible to low-dose STZ toxicity than in control rats. Under the effect of STZ, the ability of insulin secretion of beta cell is obviously decreased, while apoptosis is increased, which induces a higher glucose level and a lower insulin secretion. Administration of estrogen may protect OVX rats from the metabolic disturbances.

Objective To investigate whether the effect of pioglitazone on TNF-α-induced insulin resistance is associated with altering IRS-1-induced signaling. Methods 3T3-L1 adipocytes were treated with TNF-α for 24 hours with or without being pretreated with 10μM piglitazone for 6 hours or with pioglitazone alone.Insulin-stimulated glucose uptake of 3T3 adipocytes was measured by using 2-deoxy 3H glucose.The Western blot was used to measure IRS-1, PKB, PKC-λ protein and tyrosine phosphorylation on IRS-1, PKB and PKC-λ phosphorylation. Results Both TNF-α and pioglitazone increased glucose uptake of 3T3 adipocytes under basal status.On TNF-α treated cells, insulin-stimulated glucose uptake was decreased by about 50%, accompanied with the reductions of IRS-1 protein level, tyrosine-phosphorylation of IRS-1 and PKB phosphorylation.TNF-α treatment had no effect on PKC-λ phosphorylation. Pioglitazone pretreatment was able to antagonize TNF-α-induced insulin resistance in 3T3 adipocytes partly reverse IRS-1 protein, increase insulin-stimulated tyrosine phosphorylation of IRS-1,and increase phosphorylations of PKB and PKCλ. Conclusion TNF-α-induced insulin resistance in 3T3-L1 adipocytes is related to impaired tyrosine phosphorylation of IRS-1. Pioglitazone antagonizes the above TNF-α induced insulin resistance.

Objective:To establish the drug release determination conditions and method for phencynonate hydrochloride extended release tablets. Methods:The drug release of the tablets was determined by HPLC using a Diamonsil C18 (250 mm × 4. 6 mm, 5 μm) column with the mobile phase of acetonitrile-water-phosphoric acid-triethylamine (270∶400∶1. 3∶2), the detection wavelength was 220 nm, the flow rate was 1. 0 ml·min-1 , the column temperature was 30 ℃ and the injection volume was 20 μl. The effects of different release apparatus, release media and rotation speeds on the release of phencynonate hydrochloride extended-release tablets were studied as well. Results:The established drug release determination method had a good linear relationship within the range of 0. 3-5. 0 μg· ml-1(r=0. 999 8), and the average recovery was 100. 6%(RSD=1. 16%, n=15). Under the conditions of 900ml pH 3. 0 phos-phate buffer solution as the release medium, rotation speed of 50 r·min-1 and the settlement basket as the apparatus, the release be-havior of the product was complied with a zero-level model in vitro, and the release equation was as follows:Q=6. 141 2t-9. 328 7(r=0. 996). Conclusion:The method is simple, accurate and reliable, and suitable for the quality control of phencynonate hydrochlo-ride extended-release tablets.

Objective:To establish an HPLC method for the simultaneous determination of triamcinolone acetonide acetonide ( TAA) and chloramphenicol in the cream. Methods:The chromatographic system consisted of a ZORBAX SB-C18 column (250 mm × 4. 6 mm, 5μm). The mobile phase consisted of methanol and phosphate buffer with gradient elution at a flow rate of 1. 0 ml·min-1, the detection wavelength was 240nm, the column temperature was ambient, and the sample size was 20 μl. Results:The calibration curve was linear for TAA and chloramphenicol within the concentration range of 6. 12-48. 96 μg·ml-1 and 62. 1-745. 2 μg·ml-1 with the recovery of 99. 7% (RSD=1. 3%, n=9) and 99. 4%(RSD=1. 0%, n=9), respectively. Conclusion: The method is sensitive, accurate and specific, and can be used to control the quality of chloramphenicol and triamcinolone acetonide acetate cream.

Objective:To establish an HPLC-DAD-ELSD method for the simultaneous determination of neomycin sulfate and hy-drochloric dyclonine in compound Twaln ointments. Methods:The assay was performed on an Agilent ZOR BAXSB-C18 column(250 mm × 4. 6 mm, 5μm) with acetonitrile-water as the mobile phase with gradient elution at a flow rate of 1. 0 ml·min-1 . The detection wavelength of DAD was 282 nm. The evaporator temperature of ELSD was set at 50℃ and the nebulizer temperature was set at 60℃with the gas flow rate of 1. 6 L·min-1 . The column temperature was kept at 35℃. Results:The linear range of neomycin sulfate was 141. 54-323. 52 μg·mL-1(r=0. 999 6) with the average recovery of 98. 87%(RSD=0. 95%, n=9). The linear range of hydro-chloric dyclonine was 28. 00-64. 00 μg·mL-1(r=0. 999 6) with the average recovery of 99. 57%(RSD=1. 10%, n=9). Conclu-sion:The method is accurate, sensitive and reproducible, and under the same chromatographic conditions, the determination of all the active ingredients in compound Twaln ointments is achieved, which provides basis for the quality control.

Objective:To investigate glucose uptake and IRS-1-associated signaling pathway by stimulated insulin under TNF-? treatment.Methods:3T3-L1 adipocytes were treated with TNF-? within 6 hours and 24 hours respectively. 2-deoxy ~3H glucose was used to measure glucose uptake and western blot was used to measure IRS-1, PKB protein, tyrosine and serine307 phosphorylation on IRS-1, and PKB phosphorylation.Results:On basal status, glucose uptake of 3T3-L1 cells and phospho-tyrosine of IRS-1, PKB phosphorylation, and serine307 phosphorylation on IRS-1 were all low. Insulin stimulation induced glucose uptake and IRS-1 tyrosine phosphorylation, serine307 phosphorylation, PKB phosphorylation rapidly. TNF-? inhibited insulin-induced glucose uptake, tyrosine phosphorylation of IRS-1 and PKB phosphorylation. Rapamycin reversed the effects of TNF-?. Treated with TNF-? within 6 hours increased serine307 phosphorylation but had no effect on IRS-1 protein level. TNF-?-induced serine307 phosphorylation of IRS-1 was not affected by rapamycin. IRS-1 level was decreased under 24 hours TNF-? treatment and rapamycin can reverse the effect.Conclusion:TNF-? induced insulin resistance in 3T3-L1 adipocytes mightbe related to impaired IRS-1 tyrosine phosphorylation, rapamycin could reverse the effects of TNF-?. Treated with TNF-? within 6 hours stimulate phosphrylation of serine307 of IRS-1 and 24 hours treatment decreased IRS-1 protein level. Rapamycin antagonist TNF-?-induced loses of IRS-1.

Objective:To establish an HPLC method for the determination of chloramphenicol in chloramphenicol vaginal tablets. Methods:An Agilent ZORBAX SB-C18 column (150 mm × 4. 6 mm,5 μm)was used. The mobile phase was composed of 0. 1% sodi-um 1-heptanesulfonate solution-methanol (68∶32),the flow rate was 1. 0 ml·min-1, the UV detection wavelength was 277 nm,the column temperature was 35℃,and the injection volume was 10 μl. Results:The linear range of chloramphenicol was 25. 6-512. 0 μg ·ml-1(r=0. 999 9). The mean recovery was 99. 4%, and RSD was 0. 8%(n=9). Conclusion:The method is simple,accurate and reproducible,and can be used in the determination of chloramphenicol vaginal tablets.

OBJECTIVE:To determine the plasma concentration of Phencynonate hydrochloride(PCH)in dogs,and to calcu-late pharmacokinetic parameters. METHODS:6 Beagle dogs were given PCH tablets(2 mg and 4 mg)intragastrically. The blood samples were collected 5 min before medication and 0.17,0.25,0.5,0.75,1.0,1.25,1.5,2,3,4,6,8,10,12,14,16,18, 24 and 36 h after medication,2 ml each time. Using penehyclidine hydrochloride as internal standard,HPLC-MS/MS method was adopted to determine the plasma concentration of PCH. The medication plans were interchanged 2 weeks later. The pharmacokinetic parameters were calculated using DAS 2.0 software. RESULTS:The linear range of PCH was 0.1-15 ng/ml(r=0.999 6);the low-est limit of quantification was 0.1 ng/ml;the methodology recovery were 97.30%-103.20%;the extraction recovery were 52.30%-60.11%(RSD<11%,n=5). The main pharmacokinetic parameters of low and high doses were as follows as t1/2α of (0.678±0.525)and(0.405±0.465)h,tmax of(1.042±0.401)and(0.900±0.418)h,cmax of(14.063±6.29)and(31.580±9.673) mg/L,AUC0-36 h of(48.186±14.776)and(79.269±34.649)mg·h/L. CONCLUSIONS:The method is simple,sensitive and speci-fic,and can be used for pharmacokinetic study of PCH in dogs.

After the application of project-based learning (PBL) and 3D printing in classroom and teaching, the integrating methods and principles of PBL and 3D printing and recent teaching resources were summarized, PBL-based 3D modeling combined with recent innovative practice of 3D printing teaching model, with the course of Computer-aided medicine as an example, showed that the new teaching mode can effectively stimulate the interests of students, and cultivate their innovative thinking.

Objective To better understanding the clinical presentations of phaeohyphomycosis,and improve the diagnosis and management of the disease.Methods We reported a case of pulmonary phaeohyphomycosis caused by Exophiala jeanselmei at the First Affiliated Hospital of Guangzhou Medical University in 2008,and reviewed the relevant literature.The clinical,radio-logical and etiological features were summarized based on this case and the other 23 phaeohyphomycosis patients reported in China from January 1995 to August 2013.Results 24 Chinese cases of phaeohyphomycosis have been reported to date,including 15 males and 9 females.The age of these patients ranged from 4 to 76 (mean 40.0±21 .8)years old.Seventeen patients were otherwise healthy.The other 7 patients had complications.Clinical presentations of phaeohyphomycosis vary widely,including cutaneous and subcutaneous infection in 18 cases,pulmonary and central nervous system involvement in two cases each,para-nasal sinus and palpebral conjunctiva infection one case each.The diagnosis of 18 cases were confirmed both microbiologically and histologically.One case was confirmed histologically alone.Five cases were identified microbiologically alone.The samples for culture were collected from skin abscess (1/5 ),pulmonary tissue (2/5 ),and cervical spinal fluid (2/5 ),respectively. Twenty-two strains of causative organisms were identified,7 of which were Exophiala jeanselmei .Twenty-three patients received treatment.They were cured by antifungal agents alone (18)or in conjunction with surgical resection (4 ),or assisted with XD-635AB-based photodynamic laser therapy (1).Specifically,10 pa-tients were cured by itraconazole alone.Conclusions In China, most patients of phaeohyphomycosis have concurrent conditions or have previously received immunosuppressive agents and cor-ticosteroids.Cutaneous and subcutaneous infection were most common,located mainly on limbs,face,chest and abdominal skin.The most frequently isolated pathogen is Exophiala jeanselmei ,followed by Phialophora verrucosa and Exophiala spinifera .Itraconazole therapy would be very effective.Susceptibility testing is very useful in case of refractory infection.