OBJECTIVE To investigate the improvement mechanism of Huatan tongmai decoction on rats with polycystic ovary syndrome （PCOS） by regulating autophagy through phosphatidylinositol-3-kinase（PI3K）/protein kinase B（AKT）/mammalian target of rapamycin （mTOR） pathway. METHODS A total of 40 rats were randomly divided into blank group （purified water）， model group （purified water）， traditional Chinese medicine group [Huatan tongmai decoction， 24 g/（kg·d）] and chemical drug group [metformin， 0.16 g/（kg·d）]， with 10 rats in each group. Except for blank group， other groups were given a combination of high-fat diet and intragastric administration of 1 mg/kg letrozole suspension to establish PCOS rat model. After modeling， they were given relevant medicine or water intragastrically， once a day， for 42 consecutive days. After the last administration， the pathological and ultrastructural changes of ovarian tissue were observed. The levels of follicle stimulating hormone （FSH）， testosterone （T）， estradiol （E2） ，luteinizing hormone （LH） in serum were detected，and the LH/FSH ratio was calculated. mRNA expressions of Beclin-1， p62 and microtubule-associated protein 1 light chain 3 （LC3） in ovarian tissue were detected. The expressions of related proteins of PI3K/AKT/mTOR pathway and autophagy in rat ovarian tissues were also detected. RESULTS Compared with blank group， the pathological damage and ultrastructural changes of the ovarian tissue in the model group rats were obvious， and a large number of autophagosomes could be seen in cells. The levels of T and LH and the LH/FSH ratio in serum， as well as mRNA and protein expressions of Beclin-1 and LC3， were increased significantly （P＜0.05）， while the levels of E2 and FSH in serum， as well as mRNA and protein expressions of p62 and the phosphorylation levels of PI3K， AKT and mTOR proteins in ovarian tissue， were significantly decreased （P＜0.05）. Compared with model group， the pathological damage of ovarian tissue in the administration groups was significantly reduced， the number of autophagosomes was smaller， and the expression levels of the above indicators were significantly reversed （P＜0.05）. CONCLUSIONS Huatan tongmai decoction can inhibit autophagy in ovarian granular cells by activating the PI3K/AKT/mTOR pathway， regulate the secretion of sex hormones， alleviate pathological damage in ovarian tissues， and promote normal follicular development， thereby exerting an ameliorative effect on PCOS rats.

OBJECTIVE To explore the effects of different concentrations of aconitine on the ventricular electrophysiology of the rat heart when applied to the heart. METHODS By optical mapping technology, the effects of different concentrations of aconitine on ventricular action potential and calcium signal in rats before and 15 min after administration were observed by in vitro administration of aconitine 0.3, 1, 3 ng·mL−1. RESULTS Compared with the blank group, aconitine could be concentration-dependent to delay the conduction of action potentials under both spontaneous and 6 Hz stimulation rhythms, and there was a significant difference at a concentration of 3 ng·mL−1(P<0.05 or P<0.01). Compared with blank group, when the concentration of aconitine was 1 and 3 ng·mL−1, the action potential duration(APD) of the ventricle was significantly prolonged(P<0.01). Aconitine could also increase the dispersion of action potential conduction(P<0.05) and reduce the ratio of effective refractory period(ERP) to APD90(P<0.01). In addition, aconitine could also be concentration-dependent delay of calcium signal conduction, reduce the speed of calcium conduction(P<0.05 or P<0.01), increase the dispersion of calcium conduction and calcium transient duration(P<0.05 or P<0.01), and reduce the amplitude of calcium signal(P<0.01). CONCLUSION Using the optical labeling technique, it can be visualized that aconitine induces arrhythmia by concentration-dependent delay of ventricular action potential and calcium signaling in rats.To explore the effects of different concentrations of aconitine on the ventricular electrophysiology of the rat heart when applied to the heart. METHODS By optical mapping technology, the effects of different concentrations of aconitine on ventricular action potential and calcium signal in rats before and 15 min after administration were observed by in vitro administration of aconitine 0.3, 1, 3 ng·mL−1. RESULTS Compared with the blank group, aconitine could be concentration-dependent to delay the conduction of action potentials under both spontaneous and 6 Hz stimulation rhythms, and there was a significant difference at a concentration of 3 ng·mL−1(P<0.05 or P<0.01). Compared with blank group, when the concentration of aconitine was 1 and 3 ng·mL−1, the action potential duration(APD) of the ventricle was significantly prolonged(P<0.01). Aconitine could also increase the dispersion of action potential conduction(P<0.05) and reduce the ratio of effective refractory period(ERP) to APD90(P<0.01). In addition, aconitine could also be concentration-dependent delay of calcium signal conduction, reduce the speed of calcium conduction(P<0.05 or P<0.01), increase the dispersion of calcium conduction and calcium transient duration(P<0.05 or P<0.01), and reduce the amplitude of calcium signal(P<0.01). CONCLUSION Using the optical labeling technique, it can be visualized that aconitine induces arrhythmia by concentration-dependent delay of ventricular action potential and calcium signaling in rats.

Objective To study the efficiency and accuracy of artificial intelligence (AI) system based on fundus photograph in diabetic retinopathy(DR)screening,and evaluate the clinical application value of AI system. Methods A diagnostic trial was adopted. Total of 13683 color fundus photos were collected in Zhaoqing Gaoyao People's Hospital from March,2017 to November,2018. The AI system for DR (ZOC-DR-V1) was established,based on transfer learning + NASNet algorithm,by training 4465 precisely labeled fundus images (2510 normal,and 1955 with any stage of DR). One thousand confirmed fundus images (300 normal and 700 with any stage of DR),diagnosed by AI ( AI group ) and doctors ( 3 ophthalmologist doctors and 3 endocrinologist doctors ) ( doctor group ) , respectively. Ophthalmologist group and endocrinologist group were both composed of primary,intermediate and senior physicians. The mean reading time of each image and the total time of 1000 images were recorded. The accuracy and efficiency of AI system and doctor groups were compared. The reading process was divided into two stages. The diagnostic coincidence rate and the average reading time of each group between the two parts were calculated and compared. This study protocol was approved by Ethic Committee of Zhongshan Ophthalmic Center, Sun Yat-sen University (No. 2017KYPJ104). Results After training,the diagnostic coincidence rate of AI system (ZOC-DR-V1) in test set was 94. 7％,AUC was 0. 994. In this "man-machine to war",the diagnostic coincidence rate of primary,intermediate and senior endocrinologist was 94. 0％,91. 4％ and 93. 4％;the diagnostic coincidence rate of primary,intermediate and senior ophthalmologist was 92. 7％,94. 4％ and 95. 6％;the diagnostic coincidence rate of AI system was 95. 2％. There was no difference in the diagnostic coincidence rate between AI system and senior ophthalmologist ( P = 0. 749 ) . The mean reading time of each image of primary, intermediate and senior endocrinologists was (4. 63±1. 87),(3. 74±3. 47) and (5. 71±3. 47) seconds,and the total time of 1000 images of primary,intermediate and senior endocrinologists was 1. 29,1. 04 and 1. 58 hours;the mean reading time of each image of primary,intermediate and senior ophthalmologists was ( 7. 25 ± 6. 58 ) , ( 5. 18 ± 5. 01 ) and ( 5. 18 ± 3. 47 ) seconds,and the total time of 1000 images of primary,intermediate and senior endocrinologists was 2. 02,1. 44 and 1. 44 hours;the mean and total time of AI system was (1. 62±0. 67) seconds and 0. 45 hours,and the reading time of AI system was significantly shorter than that of the doctor groups (all at P=0. 000). The diagnostic coincidence rates between previous and posterior part of primary endocrinologist, primary and intermediate ophthalmologist were significantly different (χ2=11. 986,6. 517,10. 896;all at P<0. 05),and the mean reading time in the posterior part was significantly shorter than that in the previous part of intermediate and senior endocrinologist and primary ophthalmologist (t=4. 175,8. 189,5. 160;all at P<0. 01). While the reading time of AI system remained stable throughout the process(χ2=3. 151,P=0. 103;t=0. 038,P=0. 970). Conclusions The ophthalmic AI system based on fundus images has a good diagnostic efficiency,and its diagnostic coincidence rate can compare with senior ophthalmologist,providing a new method and platform for large-scale DR screening.

Objective To investigate personal identification of mixed seminal stain of two individuals, we combined the detection of genotyping autosomal, Y and X STR and sequencing mtDNA hypervariable Ⅰ (HV Ⅰ ) region. Methods We analyzed autosomal, Y and X STR with commercial kit and separating and sequencing HVⅠfragments of mixed seminal stain from two males by SSCP electrophoresis. Results Four genetic markers of the high amount sample can be obtained when mixed ratio is more than 1:10. When the proportion of two samples is close, the suspect could be excluded or, to some extent, identified by comparing with our results. Conclusion The combined detection of four genetic marker systems can, to some degree, solve the personal identification from mixed seminal stain of two individuals.

Objective To explore the relationship between SHP1 and CD99 expression in Hodgkin lymphoma (HL) .Methods RT‐PCR and Western blot to detect the expression of CD99 and SHP1 mRNA and protein expression in IM9、KM3、L428、L428‐CD99 cells ;fluorescence confocal to observe the expression and co‐localization of CD99 and SHP1 ;transiently interference CD99 in IM9 cells and then detect the expressing of SHP1 mRNA and protein levels .Results SHP1 mRNA was low expressed in L428、KM3 and high expressed in IM9;gene and protein expression were consistent trend;in L428‐CD99 cell SHP1 mRNA expression and protein levels were higher than L428 cell;CD99 expressed in membrane and SHP1 expressed in cell plasma;transient interference CD99 ,SHP1 decreased in mRNA expression and protein levels .Conclusion In HL cells ,SHP1 expression related to the missing expression of CD99 ,and its mechanism remains to be studied .

Objective To investigate the prothetic effect of free grafting of microvascular anastomotic peroneal artery perforator flap when used to repair the donor tissue defects of wrap-around flap of toe.Methods From January 2008 to March 2013,twenty-six cases thumb avulsion at proximal and distal phalanx level with skin and nail bed defect caused by trauma were admitted to our hospital.After incising wrap-around flap of toe to repair the thumb defects,microvascular anastomotic peroneal artery perforator flap was transplanted freely to repair the donor site of it.Results The skin flaps of 26 cases all successfully survived.After a followed-up of 3 months to 4 years,the length of donor toes were the same as before.The appearance of skin flaps were no fat and clumsy and the abradability of their skin were well.Algesia,thalposis and thigmesthesia were partially recovery.Two point discrimination reached to 5-10 mm.There were no obvious uncomfortableness in donor feet when standing and walking except wearing flip-flops.Conclusion Free grafting of peroneal artery perforator flap is a good method to repair the donor defect caused by incising wrap-around flap of toe.

OBJECTIVETo determine the optimal dosing of streptozotocin (STZ) for establishing lymphoma-bearing diabetic mouse models.

METHODSA total of 200 healthy male Balb/c mice were randomized into 4 groups (n=50) for intraperitoneal injection of a single dose of vehicle solution (control) or 75, 150, or 200 mg/kg STZ. The changes of body weight and blood glucose were observed regularly, and the success rate of modeling, mortality rate, and survival of the mice were recorded after the injections. The mice with successfully induced diabetes received subcutaneous or tail vein injection of A20 lymphoma cells, and the rate of tumorigenesis, mortality rate, and survival time were observed at 1 month and 3 months after tumor cell injection.

RESULTSCompared with the control group, the mice receiving STZ injection at 150 and 200 mg/kg showed significantly decreased body weight and increased blood glucose (P<0.05), while STZ at 75 mg/kg did not produced such obvious changes. STZ injection at 200 mg/kg resulted in a significantly higher mortality rate and shorter survival time than STZ at 150 mg/kg (P<0.05). In the control group and 150 and 200 mg/kg STZ groups, the rate of tumorigenesis or mortality rate showed no significant differences after subcutaneous injection of A20 lymphoma cells (P>0.05), but differed significantly at 3 months after tail vein injection of the tumor cells (P<0.05).

CONCLUSIONIntraperitoneal injection of STZ at 150 mg/kg is associated with a low mortality rate, a high successful modeling rate of diabetes and a long survival time in mice, and is therefore optimal for establishing diabetic mouse models bearing transplanted tumors.

Animals ; Blood Glucose ; Body Weight ; Diabetes Mellitus, Experimental ; Diabetes Mellitus, Type 2 ; chemically induced ; Injections ; Male ; Mice ; Mice, Inbred BALB C ; Streptozocin ; administration & dosage

Objective To determine the optimal dosing of streptozotocin (STZ) for establishing lymphoma-bearing diabetic mouse models. Methods A total of 200 healthy male Balb/c mice were randomized into 4 groups (n=50) for intraperitoneal injection of a single dose of vehicle solution (control) or 75, 150, or 200 mg/kg STZ. The changes of body weight and blood glucose were observed regularly, and the success rate of modeling, mortality rate, and survival of the mice were recorded after the injections. The mice with successfully induced diabetes received subcutaneous or tail vein injection of A20 lymphoma cells, and the rate of tumorigenesis, mortality rate, and survival time were observed at 1 month and 3 months after tumor cell injection. Results Compared with the control group, the mice receiving STZ injection at 150 and 200 mg/kg showed significantly decreased body weight and increased blood glucose (P<0.05), while STZ at 75 mg/kg did not produced such obvious changes. STZ injection at 200 mg/kg resulted in a significantly higher mortality rate and shorter survival time than STZ at 150 mg/kg (P<0.05). In the control group and 150 and 200 mg/kg STZ groups, the rate of tumorigenesis or mortality rate showed no significant differences after subcutaneous injection of A20 lymphoma cells (P>0.05), but differed significantly at 3 months after tail vein injection of the tumor cells (P<0.05). Conclusion Intraperitoneal injection of STZ at 150 mg/kg is associated with a low mortality rate, a high successful modeling rate of diabetes and a long survival time in mice, and is therefore optimal for establishing diabetic mouse models bearing transplanted tumors.

Objective To determine the optimal dosing of streptozotocin (STZ) for establishing lymphoma-bearing diabetic mouse models. Methods A total of 200 healthy male Balb/c mice were randomized into 4 groups (n=50) for intraperitoneal injection of a single dose of vehicle solution (control) or 75, 150, or 200 mg/kg STZ. The changes of body weight and blood glucose were observed regularly, and the success rate of modeling, mortality rate, and survival of the mice were recorded after the injections. The mice with successfully induced diabetes received subcutaneous or tail vein injection of A20 lymphoma cells, and the rate of tumorigenesis, mortality rate, and survival time were observed at 1 month and 3 months after tumor cell injection. Results Compared with the control group, the mice receiving STZ injection at 150 and 200 mg/kg showed significantly decreased body weight and increased blood glucose (P<0.05), while STZ at 75 mg/kg did not produced such obvious changes. STZ injection at 200 mg/kg resulted in a significantly higher mortality rate and shorter survival time than STZ at 150 mg/kg (P<0.05). In the control group and 150 and 200 mg/kg STZ groups, the rate of tumorigenesis or mortality rate showed no significant differences after subcutaneous injection of A20 lymphoma cells (P>0.05), but differed significantly at 3 months after tail vein injection of the tumor cells (P<0.05). Conclusion Intraperitoneal injection of STZ at 150 mg/kg is associated with a low mortality rate, a high successful modeling rate of diabetes and a long survival time in mice, and is therefore optimal for establishing diabetic mouse models bearing transplanted tumors.