OBJECTIVETo investigate the expressions of sphingosine-1-phosphate receptors 1-3 (S1P1- 3) in the corpus cavernosum of castrated male rats and its relationship with the NOS/NO/cGMP and RhoA/Rho kinase signaling pathways.

METHODSWe equally randomized 18 eight-week-old healthy male SD rats into a sham-operation control, a castration, and a testosterone replacement (TR) group and harvested the bilateral testes and epididymides from the rats in the latter two groups, followed by 4 weeks of subcutaneous injection of testosterone propionate at 3 mg per kilogram of the body weight per day for those in the TR group and that of plant oil for those in the control and castration groups. At the age of 12 weeks, we measured the serum testosterone (T) level and maximum intracavernous pressure/mean arterial pressure (ICPmax/MAP) of the animals and determined the expressions of SlP1-3, eNOS, P-eNOS, ROCK1, and ROCK2 in the corpus cavernosum by Western blot and immunohistochemistry.

RESULTSThe serum T level was significantly decreased in the rats of the castration group as compared with those of the control and TR groups ([0.41 ± 0.04] vs [16.01 ± 1.02] and [15.84 ± 1.32] nmol/L, P < 0.01), with no statistically significant difference between the latter two groups. The ICPmax/MAP at 0 V, 3 V, and 5 V electric stimulation was remarkably lower in the rats of the castration group (0.088 ± 0.014, 0.323 ± 0.014, and 0.432 ± 0.012) than in those of the control group (0.155 ± 0.011, 0.711 ± 0. 010, and 0.819 ± 0.024) and TR group (0.153 ± 0.012, 0.696 ± 0.017, and 0.763 ± 0.027) (P < 0.01), with no significant difference between the latter two groups. With GAPDH as internal control, the animals of the castration group showed markedly reduced expressions of S1P1 ([49.99 ± 3.39]%), eNOS ([46.82 ± 3.81]%) , and P-eNOS ([45.42 ± 4.35]%) in comparison with those in the control group ([72.57 ± 3.06], [89.76 ± 3.98], and [82.53 ± 8.92] and TR group ([71.77 ± 4.43], [87.19 ± 4.23], and [79.82 ± 7.38]%) (P < 0.01) , while the expressions of S1P2, S1P3, ROCK1, and ROCK2 were significantly upregulated in the castration group ([82.35 ± 4.13], [61.03 ± 5.14], [74.50 ± 4.02], and [69.83 ± 5.75]%) as compared with those in the control group ([41.67 ± 1.68], [31.66 ± 2.67], [35.69 ± 5.56], and [39.85 ± 7.17]%) and TR group ([42.80 ± 3.87], [32.25 ± 4.22], 38.06 ± 5.21], and [42.36 ± 4.44]%) (P < 0.01).

CONCLUSIONAndrogen deficiency induces significant reduction of ICPmax/ MAP in male rats, which is possibly associated with the decline of S1P1 in the corpus cavernosum, inhibition of the eNOS/NO/cGMP signaling pathway, increased expressions of S1P2 and S1P3, and activation of the RhoA/Rho kinase signaling pathway.

Animals ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Orchiectomy ; Penis ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Lysosphingolipid ; metabolism ; Testosterone ; blood ; pharmacology ; rho-Associated Kinases ; metabolism

Objectives To investigate the role of laminin (LM) and fibronectin (FN) in the growth cycle of human hair follicles. Methods The expression of LM and FN was detected by streptavidin-peroxidase (SP) staining. Results In the anagen phase, LM was expressed in dermal papilla, basement membrane and outer root sheath; FN was expressed in dermal papilla, basement membrane and connective tissue sheath. In the catagen phase, LM showed a lower expression in the dermal papilla and a linear expression in the basement membrane; the expression of FN in the dermal papilla and basement membrane was less intense than that in anagen phase, but was still positive. In the telogen phase, LM was only expressed in the basement membrane while FN was negative. Conclusion The difference between LM and FN expression in hair growth cycle indicates that LM and FN may play important roles in the regulation of human hair follicle growth cycl.

Objective To analyze slowly the digestibility and postprandial glycemic response after consum of different cereal starch. Method Starch was quantified into different nutritional fractions using the in vitro Englyst test. Ten healthy subjects consumed 7 kinds of carbohydrate foods with 50 g normal maize starch, waxy maize starch, wheat starch, sticky rice starch, rice starch, potato starch or glucose. Blood samples from postprandial zero to 120 min after consumption of test materials were collected for measurement of glucose, glycemic index and extended glycemic index. Results Native cereal starch was ideal slowly digestible starch (SDS) and the proportion of SDS was about 50% based on Englyst test. The GI value of cereal starch was more than 90% and belonged to high glycemic index foods, but EGI was positive, regarding glucose powder as 100%. Conclusion Cereal starch with slow digestibility and similar glycemic response was a better carbohydrate material to provide a slow and prolonged release of blood glucose and maintenance of glucose homeostasis, which was potentially beneficial to health.

Background and purpose: Lysine specific demethylase 1(LSD1) is an important chromatin modifier. It epigenetically regulates gene expression pattern through chromatin modification and participates in maintenance of tumor malignant properties, such as oncogenesis, development, invasion, migration and metabolic transformation. SIRT3 (sirtuin 3) is a mitochondria localized tumor suppressor and regulates tumor metabolic transformation and oxidative stress. The correlation between LSD1 and SIRT3 has never been reported before. This study aimed to elucidate the correlation between LSD1 and SIRT3 with gene transcriptional regulation methods. Methods: RNA interference technique, co-immunoprecipitation assay(CoIP), chromatin immune-precipitation assay(ChIP) and ifrelfy luciferase activity assay were employed to elucidate the correlation between LSD1 and SIRT3 in pancreatic cancer. Results:mRNA and protein levels of SIRT3 were signiifcantly elevated in LSD1 knock-down PANC-1 cells. LSD1 interacts with PGC-1α, an important regulator of SIRT3 gene expression. LSD1 and PGC-1αoccupied the same region in SIRT3 promoter region through ChIP analysis. Luciferase activity assay validated LSD1 as a negative regulator of PGC-1αin SIRT3 gene transcriptional regulation. Conclusion:LSD1, as an important tumor promoter, negatively regulates the expression of tumor suppressor gene SIRT3, these results provide important clues for the role that LSD1 plays in aberrant metabolism and oxidative stress.

AIM: To observe the therapeutic effect of combined trabeculectomy with improved conjunctival incision at 1mm of corneoscleral limbus posterior
METHODS: A retrospective analysis of 171 cases ( 220 eyes ) with primary glaucoma who were randomly divided into 3 groups according to the differences of conjunctival incision and the way of suture. Group A:44 eyes of 33 cases underwent corneoscleral limbus conjunctival incision; Group B: 94 eyes of 76 cases underwent conjunctival incision at 1mm of corneoscleral limbus posterior; Group C: 82 eyes of 62 cases underwent conjunctival flap running suture based on group B. All cases were followed up for 1a. Intraocular pressure, morphology of filtering bleb, depth of anterior chamber and water leakage in conjunctival incision were observed and compared after operation among three groups.
RESULTS: The incidence of postoperative water leakage inconjunctival incision and shallow anterior chamber were group A >group B>group C and there were significant differences among the three groups ( P<0.05), The success rate was 94. 5%. The intraocular pressure and morphology of bleb had no significant differences among the three groups.
CONCLUSION: Conjunctival incision at 1mm of corneoscleral limbus posterior and conjunctival flap running suture can not only reduce the rate of water leakage in conjunctival incision and accelerate the speed of incision adhesion after operation, but also significantly enhance the therapeutic effect of combined trabeculectomy.

Objective To detect the serum levels of 25‐hydroxy vitamin D[25‐(OH)D] ,homocysteine(HCY) andβ2‐microglob‐ulin(β2‐MG) in the patients with type 2 diabetes mellitus (T2DM ) and to investigate the relationship between serum HCY and β2‐MG with 25‐(OH)D and its clinical significance .Methods A total of 139 cases of T2DM were selected anddivided into 3 groups , the normal albuminuria group for [urinary albumin to creatinine ratio (UACR)< 30 mg/gCr ,45 cases] ,microalbuminuria group (UACR ≥ 30 mg/gCr and < 300 mg/gCr ,48 cases) and massive proteinuria group (UACR ≥ 300 mg/gCr ,46 cases) according to the urinary albumin to creatinine ratio (UACR) .Other 45 individuals undergoing the physical examination were selected as the con‐trol group .The serum 25‐(OH)D level was measured by electrochemiluminescence .Serum HCY level was determined by the enzy‐matic method .Serum β2‐MG level was measured by the latex enhanced immune turbidity method .At the same time ,the biochemical indicators of FBG ,HbA1C ,serum calcium and phosphorus were measured .Results The serum 25‐(OH)D level was decreased with the increase of urinary albumin in the DM patients .And the serum 25‐(OH)D level in the microalbuminuria group and the massive proteinuria group was significantly decreased compared with the normal albuminuria group and the control group ,the difference was statistically significant(P<0 .01) .The serum HCY and β2‐MG levels in the microalbuminuria group and the massive proteinuria group were significantly increased compared with the healthy control group ,the difference was statistically significant (P<0 .05) . Conclusion The serum 25‐(OH)D level is decreased with the increase of urinary albumin in the diabetic patients .The serum HCY andβ2‐MG levels are increased with the increase of urinary albumin and serum 25‐(OH)D level is negatively correlated with the HCY andβ2‐MG levels .

Objective: To observe the therapeutic efficacy of astragalus polysaccharide (APS) in combination with three-dimensional conformal radiotherapy (3D-CRT) for elderly patients with non-small cell lung cancer (NSCLC). Methods: A total of 80 elderly patients with I～IV stage NSCLC were randomly divided into two groups. The 40 patients in the therapeutic group received radiotherapy in combination with APS. The 40 patients in the control group received radiotherapy alone. Both groups received 3D-CRT with a total dose of 50～ 70Gy, 2.0Gy/fraction, once a day, 5 times per week. The patients in the therapeutic group were treated with radiotherapy combined with injection of APS (250mg in 5% glucose) or normal saline (500 mL) intravenously once a day until the end of radiotherapy. The short-term efficacy and patients' quality of life were evaluated. The T-lymphocyte subpopulation and peripheral blood leukocyte count were also measured after treatment. Results: The short-term effective rates of the therapeutic group and the control group were 87.5% and 72.5%, respectively, without significant difference between the two groups (P＞0.05). The decrease of peripheral blood leukocyte count after treatment in the control group was significant (P＜0.05). In the therapeutic group, the T-lymphocyte subsets CD8 and CD4/CD8 were improved after treatment, with a significant difference (P＜ 0.05). But in the control group, no significant changes in T-lymphocyte subsets CD8 and CD4/CD8 were observed after treatment (P＞0.05). Patients' quality of life in the therapeutic group was superior to that in the control group, with a significant difference (P＜0.05). Conclusion: APS in combination with 3D-CRT can reduce the side effects of radiation and improve the quality of life of elderly patients with NSCLC.

BACKGROUND: Because the continuity and integrity of the trachea are likely damaged to some extent after tracheostomy, the implementation of sequential ventilation has certain difficulties, and sequential invasive-noninvasive ventilation on patients after tracheostomy is less common in practice. The present study aimed to investigate the feasibility of invasive-noninvasive sequential weaning strategy in patients after tracheostomy. METHODS: Fifty patients including 24 patients with withdrawal of mechanical ventilation (conventional group) and 26 patients with sequential invasive-noninvasive weaning by directly plugging of tracheostomy (sequential group) were analyzed retrospectively after appearance of pulmonary infection control (PIC) window. The analysis of arterial blood gases, ventilator-associated pneumonia (VAP) incidence, the total duration of mechanical ventilation, the success rate of weaning and total cost of hospitalization were compared between the two groups. RESULTS: Arterial blood gas analysis showed that the sequential weaning group was better than the conventional weaning group 1 and 24 hours after invasive ventilation. The VAP incidence was lowered, the duration of mechanical ventilation shortened, the success rate of weaning increased, and the total cost of hospitalization decreased. CONCLUSION: Sequential invasive-noninvasive ventilator weaning is feasible in patients after tracheostomy.

OBJECTIVE: To investigate the effect of miR-590-5p on the expression of lectin-like oxidized low density lipoprotein receptor 1 (LOX-1) in apoptotic human umbilical vein endothelial cells (HUVECs) induced by ox-LDL, and to explore the role of miR-590-5p in modulating HUVECs apoptosis. METHODS: HUVECs were exposed to ox-LDL (50 μg/mL) for 0 to 48 h. Apoptosis was detected by Annexin V-FITC stain and was distinguished from necrosis by propidium iodide (PI) staining. The relative expression level of miR-590-5p in HUVECs was analyzed using real-time quantitative PCR (RT-qPCR). HUVECs were transfected with miR-590-5p mimics or miRNA mimics control followed by 50 μg/mL ox-LDL stimulation for 48 h. LOX-1 mRNA and protein were measured by RT-qPCR and Western blot, and apoptosis in HUVECs was analyzed by flow ctyometry after Annexin V-FITC/PI double stain. RESULTS: Incubation of HUVECs with 50 μg/mL ox-LDL for 0 to 48 h resulted in a time-dependent induction of apoptotic cell death and down-regulation of miR-590-5p. Transfection of miR-590-5p mimics suppressed LOX-1 expression at both mRNA and protein levels, leading to a reduction of ox-LDL-induced apoptosis in HUVECs. CONCLUSION MiR-590-5p protects endothelial cells from ox-LDL induced apoptosis by inhibiting the expression of LOX-1.
Apoptosis ; genetics ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Lipoproteins, LDL ; pharmacology ; MicroRNAs ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Scavenger Receptors, Class E ; genetics ; metabolism ; Transfection

ObjectiveTo investigate the survival and differentiation of autologous bone-marrow mononuclear cell (ABM-MNCs) after transplanted to infarcted area and border area, and the effect of ABM-MNC on the cardiac function.Methods40 male big-ear Japanese rabbits were divided randomly into the transplanted group and control group with 20 animals in each group. Acute myocardial infarction model was made by ligating left anterior descending artery. 7 days later, Brdu labeled ABM-MNCs were injected into myocardium in the transplanted group, while the control rabbits were injected with saline. Six weeks later, tests of histology and immunohistochemistry were performed.ResultsViable cells labeled with Brdu can be identified in the infarcted area, and myocytes and endothelial cells labeled with Brdu can also be found in the border area, these cells demonstrated myogenic differentiation with the expression of α-actin by immunostaining. While, no cells labeled with Brdu were found in the control group. Moreover, the vessel density of the transplanted group in the borders of the infarction was higher than the control group (P<0.05), but there was no difference in infarcted area between two groups (P>0.05).At the 6 weeks after experiment, the cardiac function was improved in both groups, but there was a significant difference between two groups (P<0.05).ConclusionABM-MNCs injected into the infarcted myocardium can survive in both the infarcted and border areas, and differentiate into endothelial cells and other cells which are able to obtain the characters of myocytes, and increase the vessel density in border area, improve the cardiac function.