[Neuroprotective effect of ginsenoside Rb-1 on a rat model of Alzheimer's disease].

Objective: To investigate the protective mechanism of ginsenoside Rb-1 on the brain in a rat model of Alzheimer's disease. Methods: Fifty-six male Sprague-Dawley rats were randomly divided into control group, model group, low-dose Rb-1 group (Rb-1: 25 mg•kg(-1)•d(-1)) and high-dose Rb-1 group (Rb-1:50 mg•kg(-1)•d(-1)). Morris water maze was designed to observe the changes of learning and memory ability in rats. Flow cytometry was used to detect the apoptosis of hippocampal neurons. Immunohistochemistry and Western blot were employed to detect the expression levels of apoptosis-related genes (p53, Bax, cytochrome C (Cyto C), Caspase-3 and caspase-9) and anti-oxidative stress-associated genes (nuclear Factor-E2-related factor 2 (Nrf2), kelch-like ECH-associated protein 1 (keap-1), heme oxygenase 1(HO-1) and NADPH quinone dehydrogenase 1 (NQO1)).The activities of catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were detected by relevant kits. ANOVA and Tukey-Kramer test were used for statistical analysis. Results: The learning and memory ability of rats in the model group was lower than that of the control group (P<0.01).The learning and memory ability of rats in the high-dose Rb-1 treatment group was significantly higher than that of the model group [(80±8) s vs (100±11) s, t=5.390, P<0.01]. The expression levels of apoptosis-related genes (p53, Bax, Cyto C, caspase-3 and caspase-9) in the model group were significantly higher than those in the control group (P<0.01), while the expression levels of these genes in low-dose and high-dose Rb-1 groups were significantly lower than those of the model group (P<0.01). The expression levels of Nrf2, HO-1 and NQO1 genes in the model group were significantly lower than those in the control group (P<0.05), while the expression of these genes in low-dose and high-dose Rb-1 groupswere significantly higher than those of the model group (P<0.01). The activities of CAT, GSH-Px and SOD in the model group were lower than those in the control group (P<0.01), however the activities of CAT, GSH-Px and SOD in low-dose and high-dose Rb-1 groups were higher than those of model group (P<0.05). Conclusions: Both low-dose and high-dose Rb-1 have protective effect on memory and cognitive function of Alzheimer's disease rats by reducing the damage and apoptosis of hippocampal neurons, down-regulating the expression levels of p53, Bax, Cyto C, caspase-3 and caspase-9, up-regulating the expression of Nrf2, HO-1 and NQO1 genes, and increasing the activities of CAT, GSH-Px and SOD. Moreover, the protective effect of Rb-1 on rat brain may be dose-dependent.

[Preliminary investigation of gender assignment in 46,XY disorders of sex development with severe male undermasculinisation].

Objective: To explore the feasibility of gender assignment in 46,XY disorders of sex development (DSD) with severe undermasculinisation mainly based on molecular diagnosis. Methods: A retrospective study of 45 patients of 46, XY DSD with severe undermasculinisation were admitted between November 2015 and October 2018 at Children's Hospital, Zhejiang University School of Medicine. The initial social gender were all female, of whom the external genital manifestations were Prader 0 to 2; the degree of masculinity was scored using external masculinisation score (EMS); the position and development of the gonads were examined by ultrasound, cystoscopy and laparoscopy, also including assessing the development of the Wolffian tube and the Müllerian tube. The level and ratio of testosterone to dihydrotestosterone before and after hCG stimulation were evaluated for the function of Leydig cell and 5α-reductase-2. Gender role scales and sandbox games were used to assess gender role behavior. Genital sensitivity to androgen stimulation was assessed; A panel including 163 genes related to gender development were determined by second-generation sequencing in all 45 patients. Finally, a multidisciplinary team (MDT) makes a gender assignment after a comprehensive analysis mainly based on the molecular etiological diagnosis. Results: Thirty-nine out of 45 patients (87%) had an identifiable genetic etiology, and the remaining 6 (13%) were negative for genetic testing. Forty-five patients had EMS less than or equal to 3 points. Sexual psychological assessment was performed in 39 patients, with male dominance in 24 (62%) and female dominance in 15 (38%). The gender assignment was 23 cases (51%) for male and 19 cases (42%) for female, and 3 cases (7%) were not completely determined. Conclusions: Molecular diagnosis provides a strong basis for appropriate gender assignment of 46, XY DSD children with severe undermasculinisation. Based on molecular diagnosis, each DSD should be analyzed by professional MDT to analyze the clinical symptoms/signs, gonadal development, gonad tumor risk, external genital morphology, sexual psychological assessment, potential fertility opportunities, parental views, Social and cultural factors, etc. make appropriate gender assignment.

[One patient with chronic refractory wound caused by bamboo sticks in lower leg].

One patient with chronic refractory wound in right lower leg was admitted to our department in June 2017, which experienced repeated ulceration for 4 years. On 4 days post admission, two bamboo sticks were taken out from the deep muscle of the right lower leg. The wound was repaired by perforator flap of posterior tibial artery in the right lower leg. The donor site was covered with split-thickness skin in the upper leg of the same side. On 7 days post operation, the flap and skin graft survived. During follow-up of 12 months, the flap had good appearance. This case suggests that the first thing for the treatment of chronic refractory wound is to find out the cause of wound.

[Effect of inhibiting perivascular adhesion on intimal hyperplasia of the vein grafts in rabbits].

Objective: To investigate the effect of preventing perivascular adhesion with topical application of sodium hyaluronate on intimal hyperplasia of the vein grafts in rabbits. Methods: Twenty-four male New Zealand white rabbits, aged 5 months, were randomly divided into 2 groups: Group A and B (n=12 rabbits per group). Artery defect model was established by cutting about 1 cm artery from the middle part of the dissociated left common carotid artery. A section about 3 cm was cut from the right external jugular vein, and the harvested vein was inverted and end-to-end anastomosed to the artery defect. After anastomosis, the adventitia and two anastomosis of the grafted veins in group A was applied 0.2 ml sodium hyaluronate locally to, and corresponding site in Group B was served as a control, but with the sterile normal saline. The grafted veins were obtained 1, 2 and 4 weeks after operation, HE staining and Masson staining were preformed for histological changes of grafted vein wall, proliferating cell nuclear antigen (PCNA) and platelet-derived growth factor (PDGF) immunohistochemistry staining were conducted for proliferation and expression and distribution of PDGF of the grafted vein. Results: The macroscopic and histological observation showed that the perivascular adhesions in Group A were looser when compared with those in Group B. The thickness of the intima, the degree of intima hyperplasia of 2 groups at different time points were as follows: at 1 week after operation, group A[(25.5±3.9) µm, (1.2±0.1) ]and group B[(26.2±4.2)µm, (1.2±0.1)]; at 2 weeks after operation, group A[(44.3±2.5)µm, (1.2±0.1)]and group B[(51.0±3.8)µm, (1.4±0.0)]; at 4 weeks after operation, group A[(69.9±6.8)µm, (1.5±0.1)] and group B[(84.4±6.4)µm, (1.7±0.1)]. Group A was inferior to group B in terms of the above three parameters 2 and 4weeks after operation (P<0.05). Cell proliferation index of intima and that of media were as follows: at 1 week after operation, group A (7.4±2.2), (21.5±3.2) and group B (11.5±2.0), (28.6±4.5); at 2 weeks, group A (20.0±3.2), (35.8±3.4) and group B (26.8±4.1), ( 42.6±4.2); at 4 weeks, group A (11.4±2.0), (22.1±2.7) and group B (15.5±2.4, 28.6±3.9). Group A was inferior to group B in terms of cell proliferation index of intima and media 1, 2 and 4 weeks after operation (P<0.05). The percentage of PDGF-positive cells of intima, media and adventitia was as follows: at 1 week after operation, group A (7.7±1.6), (19.6±3.7), (2.5±1.5) and group B (7.6±2.4), (20.6±4.4), (10.3±2.3); at 2 weeks after operation, group A (11.4±2.6), (19.8±3.1), (12.9±3.3) and group B (19.5±3.5), ( 30.6±5.2), (30.5±5.8); at 4 weeks after operation, group A (6.2±1.9), ( 11.1±2.8), (10.2±2.4) and group B (10.5±2.0), (18.6±3.2), (26.5±3.8). Group A was inferior to group B in terms of the percentage of PDGF-positive cells of intima, media and adventitia 2 and 4 weeks after operation (P<0.05) and Group A was inferior to group B that of adventitia 1 week after operation (P<0.05). Conclusion: Preventing perivascular adhesion with topical application of sodium hyaluronate can inhibit intimal hyperplasia.

Association of the kappa-opioid system with alcohol dependence.

Opioid receptors and their endogenous peptide ligands play important roles in the reward and reinforcement of drugs such as heroin, cocaine, and alcohol. The binding of dynorphins to the kappa-opioid receptor has been shown to produce aversive states, which may prevent the development of reinforcement. We genotyped SNPs throughout OPRK1, encoding the kappa-opioid receptor, and PDYN, which encodes its ligand prodynorphin, in a group of 1860 European American individuals from 219 multiplex alcohol dependent families. Family-based analyses demonstrated associations between alcohol dependence and multiple SNPs in the promoter and 3' end of PDYN, and in intron 2 of OPRK1. Haplotype analyses further supported the association of PDYN. Thus, variations in the genes encoding both the kappa-opioid receptor and its ligand, OPRK1 and PDYN, are associated with the risk for alcohol dependence; this makes biological sense as variations in either should affect signaling through the kappa-opioid system.