Effect and mechanisms of Polygonatum kingianum (polygonati rhizome) on wound healing in diabetic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic dermatopathy is one of the most serious and common complications of diabetes. It has been found that high glucose can lead to abnormal glycometabolism. The skin microenvironment pollution caused by the increase in glucose and the oxidative stress mediated by the deposition of advanced glycation end products can lead to invisible skin injury, and the interaction between them is the key factor that makes the skin wounds of diabetic rats difficult to heal. Therefore, the main task of promoting healing is to reduce blood glucose levels and relieve the deposition of advanced glycation end products. Polygonatum kingianum Collett & Hemsl (PK) of Asparagaceae is planted in Yunnan, China, and is used by the Bai, Hani and Wa nationalities as a traditional medicine for preventing and treating diabetes. AIM OF THE STUDY: To study the effects of PK extract on skin wound healing in diabetic rats and to explore the regulatory mechanism of PK on wound microenvironment pollution, the antioxidative stress signaling pathway and latent injury of wound skin tissue. METHODS: First, wounds were prepared after diabetic rats were given PK extract by gavage for 4 weeks, and then gavage was continued for 2 weeks to observe and calculate the wound healing rate. A scanning electron microscope was used to observe the pathomorphological changes in the skin tissue at the edge of the wound. Western blotting was used to detect protein expression. Immunohistochemistry was used to detect the expression of CD34, AGEs, bFGF and VEGF. The Nrf2/HO-1 signaling pathway in skin tissue was detected by fluorescence quantitative PCR. Serum biochemical indicators and inflammatory cytokine levels were detected by a kit. RESULTS: After PK treatment, the wound healing rate increased significantly (P < 0.001), the infiltration of inflammatory cells in skin tissue of DM lesion rats decreased, the number of new blood vessels increased, and the epidermis and dermis thickened. The content of glucose, AGEs, RAGE protein and RAGE mRNA in skin decreased significantly (P < 0.05, P < 0.01, P < 0.001), while the expression of Nrf2 mRNA, HO-1 mRNA, CD34, bFGF and VEGF increased significantly (P < 0.05, P < 0.01, P < 0.001). The levels of SOD, GSH, MMP-9 and MMP-2 in skin decreased (P < 0.05, P < 0.01, P < 0.001), but the level of TIMP-2 increased (P < 0.001). GSP, GHb and ICAM-1 in plasma decreased (P < 0.05, P < 0.01, P < 0.001), while T-AOC, SOD and FINS increased (P < 0.05, P < 0.01). The levels of MDA, TNF-, IL-6, IL-2 and IFN-γ in plasma and wound skin tissue decreased (P < 0.05, P < 0.01, P < 0.001). CONCLUSION: PK can reduce the infiltration of inflammatory cells and glucose content in the skin tissue at the edge of the wound, reduce inflammatory factors in skin and plasma, and increase angiogenesis, thus improving the wound healing rate. PK can alleviate the microenvironment pollution caused by AGEs and glucose metabolism disorder in diabetic rats and induce antioxidant activity through the Nrf 2/HO-1 signaling pathway, thus reducing oxidative damage and offsetting endogenous skin damage and hidden damage.

Effects of ginsenoside Rg3 on epidermal cells and angiogenesis in impaired diabetic wound rats / 中国药理学通报

Aim: To explore the repair mechanism of ginsenoside Rg3 in treating the impaired diabetic wound from improving the vulnerability of diabetic skin of rats. Methods: Male SD rats (n = 32) were injected intraperitoneally streptozotocin to induce hyperglycemia, except for the rats of control group (n = 8). Diabetic rats were randomly divided into model group (equal volume of saline), aminoguanidine group (10 mg · kg-1), ginsenoside Rg3 high dose (15 mg · kg-1) and low group (5 mg · kg-1). After four weeks of oral gavage treatment, full-thickness wound was established. On 21st day after injury, wound samples were collected to observe the pathological changes in wound; the thickness of epidermis and dermis was measured by HE staining; and the epidermal cell proliferation cycle was analysed by flow cytometry; the expression of CD31 in wound tissues was detected by immunohistochemical and image analytical methods. Results: Ginsenoside Rg3 groups showed significantly more fibroblasts, necrotic tissue drops and advanced epithelial, and thickening of epidermis and dermis (P < 0. 01). Model group showed significant increase in G0/G1 phase ratio of epidermal cell cycle (P <0. 01), while reduction in G2/M and S period ratios (P < 0. 01). However, G0/G1 period ratio decreased, while G2/M and S period ratios rose in aminoguanidine group, ginsenoside Rg3 high dose and low dose groups. The decrease of G0/G1 period ratio (P < 0. 05) and increase in G2/M (P <0. 05) and S period ratios (P <0. 01) was found to be significant. The expressions of CD31 in model group was lower than those in control group (P<0. 01). Whereas, it was higher in ginsenoside Rg3 high dose group and aminoguanidine group (P < 0. 05). Conclusions: Ginsenoside Rg3 can effectively promote the repair and healing of impaired diabetic wound. The various mechanisms of repair might be through improving skin pathology, regulating epidermal cell proliferation cycle and promoting angiogenesis.

Immunoregulation on Mice of Low Immunity and Effects on Five Kinds of Human Cancer Cells of Panax japonicus Polysaccharide.

The goal of this study is to investigate the immunoregulative effects of Panax japonicus polysaccharide (PJPS) on mice of low immunity. An orthogonal experiment was designed to determine the best extraction process for PJPS. By the tests of macrophages swallow chicken red blood cells, Delayed-type hypersensitivity (DTH), and serum hemolysin value, we studied the immune adjustment ability of PJPS. MTT was employed to detect the effects of different concentrations of PJPS, respectively, in 24 h, 48 h, and 72 h on five kinds of human cancer cells. The results show that the best extraction process for PJPS was as follows: ratio of solvent consumption to raw material 40, extraction temperature 100°C, re-extracted two times, each extraction time 4 hours. PJPS can significantly improve the immune function of mice processed by cyclophosphamide and PJPS did not work on the above five cancer cells.

Microsurgical treatment of obstructive azoospermia: a report of 76 cases / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the clinical effect of microsurgical vasoepididymostomy and/or vasovasostomy in the treatment of obstructive azoospermia.</p><p><b>METHODS</b>This study included 76 patients with obstructive azoospermia, 53 treated by bilateral vasoepididymostomy (8 involving the epididymal head, 18 involving the epididymal body, 5 involving the epididymal tail, and 22 involving the epididymal head, body and tail), 14 by unilateral vasoepididymostomy, and the other 9 by unilateral vasoepididymostomy + unilateral vasovasostomy (including cross anastomosis). We followed up the patients for 2 to 16 months for the patency rate, routine semen parameters, and pregnancy outcomes.</p><p><b>RESULTS</b>The success rate of bilateral vasoepididymostomy, unilateral vasoepididymostomy, and unilateral vasoepididymostomy + unilateral vasovasostomy (including cross anastomosis) were 62.26% (33/53), 35.71% (5/14), and 77.78% (7/9), respectively. The average sperm concentrations in the three groups of patients were (27.9 +/- 5.74), (11.8 +/- 8.33), and (19.9 +/- 7.53) x 10(6)/ml, the average total sperm counts were (65.6 +/- 13.71), (28.0 +/- 15.86), and (69.2 +/- 28.59) x 10(6), and the mean rates of progressively motile sperm were (22.3 +/- 3.18), (11.0 +/- 9.77), and (15.8 +/- 5.05)%, respectively. The success rates of bilateral vasoepididymostomy that involved the epididymal head, body, tail, and all the three parts were 62.5, 72.22, 60, and 54.55%, respectively. Natural pregnancy was achieved in 8 (10.53%) of the total number of cases.</p><p><b>CONCLUSION</b>Microsurgery is effective for the treatment obstructive azoospermia. Unilateral vasoepididymostomy + unilateral vasovasostomy is superior to the other procedures, followed by bilateral vasoepididymostomy. Bilateral vasoepididymostomy involving the epididymal body may achieve a slightly better effect than that involving the other epididymal parts.</p>

Establishing a mouse model of Sertoli-cell-only syndrome by administration of busulfan / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish a stable and reliable model of Sertoli-cell-only syndrome in mice.</p><p><b>METHODS</b>We randomly divided 60 NIH mice into two groups of equal number to receive intraperitoneal injection of busulfan (30 mg/kg) and 30 or 60 minutes of testis cooling. At 2, 4 and 8 weeks after treatment, we recorded the survival rate of the mice, weight of the testis and Johnsen scores, and conducted quantitative analysis on the degrees of spermatogenetic failure.</p><p><b>RESULTS</b>There were no significant differences in the baseline body weight and survival rate between the intervention and control groups (P > 0.05). At 4 and 8 weeks, the testis weight and Johnsen score were significantly lower in the intervention group than in the control ([0.04 +/- 0.01] g and [0.05 +/- 0.01] g vs [0.09 +/- 0.03] g and [0.11 +/- 0.02] g, P < 0.05; 3.86 +/- 0.50 and 2.70 +/- 0.67 vs 9.60 +/- 0.25 and 9.76 +/- 0.43, P < 0.01). At 2, 4 and 8 weeks, the testis weights were (0.07 +/- 0.02) g, (0.06 +/- 0.01) g and (0.09 +/- 0.01) g, respectively, in the 30-min cooling group and (0.05 +/- 0.01) g, (0.04 +/- 0.02) g and (0.04 +/- 0.02) g in the 60-min cooling group, significantly lower than in the control side at the same time points ([0.11 +/- 0.01] g, [0.11 +/- 0.01] g and [0.12 +/- 0.00] g) (P < 0.05), and the Johnsen scores were 4.70 +/- 0.67, 2.70 +/- 0.84 and 6.10 +/- 1.14 in the 30-min and 1.67 +/- 0.58, 1.20 +/- 0.45 and 1.00 +/- 0.00 in the 60-min cooling group, remarkably lower than in the control side (9.60 +/- 3.23, 9.60 +/- 0.55 and 9.70 +/- 0.45) (P < 0.01). Histopathological examination of the cooled testes revealed considerable atrophy of seminal tubules, necrosis of seminiferous epithelia and peritubular fibrosis.</p><p><b>CONCLUSION</b>Administration of busulfan has no obvious influence on the survival of mice, and is a reliable method for constructing a mouse model of Sertoli-cell-only syndrome.</p>

DAZL gene polymorphisms and astheno-teratozoospermia / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the association between single nucleotide polymorphism (SNP) of the DZAL gene in infertile Han Chinese males with astheno-teratozoospermia.</p><p><b>METHODS</b>We collected semen samples from 173 infertile Han Chinese men with astheno-teratozoospermia (case group) and 175 age-matched normal male volunteers (control group) for semen routine and morphological analyses. We obtained genomic DNA, genotyped the polymorphisms of the DAZL gene A260G and A386G via the Sequenom MassARRAY system, and compared the frequencies of the genotypes between the case and control groups.</p><p><b>RESULTS</b>The AA nucleotide variant was found in the A260G and A386G polymorphisms of the DZAL gene in both the cases and controls, but the heterozygous AG variant in neither.</p><p><b>CONCLUSION</b>The A260G and A386G polymorphisms of the DAZL gene are not correlated with astheno-teratozoospermia-induced male infertility in the Han Chinese population, and therefore could not be considered as molecular markers of male infertility.</p>

Efficacy of low-dose tadalafil on ED assessed by Self-Esteem and Relationship Questionnaire / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of low-dose oral tadalafil on self-esteem, confidence and sexual relationship in ED patients.</p><p><b>METHODS</b>We treated 17 ED patients with oral tadalafil at the low dose of 5 mg once daily for 12 weeks, and used the paired t test to compare their scores on The Self-Esteem and Relationship Questionnaire (SEAR) and IIEF-5 and the results of nocturnal penile tumescence (NPT) obtained by nocturnal electrobioimpedance volumetric assessment (NEVA) before and after the medication.</p><p><b>RESULTS</b>The scores on SEAR and IIEF-5 were significantly increased (P < 0.01) and NPT markedly improved (P < 0.05) after tadalafil treatment as compared with the baseline.</p><p><b>CONCLUSION</b>Low-dose oral tadalafil once daily can significantly improve the self-esteem, confidence, sexual relationship satisfaction and NPT of ED patients.</p>

Effects of 17beta-estradiol on alkaline phosphatase and osteoprotegerin expression in human periodontal ligament cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of 17beta-estradiol on the expression of alkaline phosphatase (ALP) and osteoprotegerin in human periodontal ligament cells.</p><p><b>METHODS</b>Human periodontal ligament cells (hPDLC) were obtained from periodontal tissue explants of teeth extracted for orthodontic treatment ALP activity was determined by PNPP, and OPG protein and corresponding mRNA levels were quantitatively detected by ELISA and RT-PCR RESULTS: ALP activity was significantly increased at 14 days and 21 days (P <0.05). 17beta-E2 of physiological concentration promoted secretion of OPG protein and expression of OPG mRNA (P <0.05). 17beta-E2 with high-dose showed no effect on OPG protein secretion and decrease OPG mRNA expression.</p><p><b>CONCLUSIONS</b>17beta-E2 may have a positive impact on periodontium through promoting expression of ALP and OPG in hPDLC.</p>

Insulin-like growth factor-II and basic fibroblast growth factor affect periodontal ligament cells expressing osteoprotegerin in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study was carried out to investigate the effects of insulin-like growth factor-II (IGF-II) and basic fibroblast growth factor (bFGF) on osteoprotegerin (OPG) secretion of periodontal ligament cells (PDLCs).</p><p><b>METHODS</b>Healthy human premolars extracted for orthodontic reasons from 12-14 years old donators were obtained, and periodontal tissues were collected and cultured to obtain PDL cells. Primary or first passage PDLCs were cloned by means of limited dilutions. PDLCs with osteoblastic phenotypes were characterized as follows: Alkaline phosphatase activity, collagen III production and bone-like nodules formation. IGF-II and bFGF were added into culture media and their effects on PDLCs proliferation and OPG secretion were observed. The OPG concentrations in cell culture supernatants were detected by sandwich ELISA. Living cell numbers were demonstrated by MTT test. The average levels of OPG secretion by a single cell were calculated by dividing OPG concentration with MTT-test result.</p><p><b>RESULTS</b>Both IGF-II and bFGF upregulated the mtt values (P < 0.05), but ICF-II downregulated the opg/mtt values (P < 0.05), whereas bFGF had no significant effect on opg/mtt values (P > 0.05).</p><p><b>CONCLUSION</b>IGF-II enhances the proliferation of PDL cells but prohibits OPG secretion. Although bFGF has the same effect on the proliferation of PDL cells, it has no effect on OPG secretion. Before cytokines were used to enhance periodontal regeneration, their effects on local bone balance should also be studied.</p>

Tumor necrosis factor and lipopolysaccharide affect periodontal ligament cells expressing osteoprotegerin in vitro / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of LPS and/or TNF-alpha on periodontal ligament cell (PDLC) proliferation and OPG secretion.</p><p><b>METHODS</b>Healthy premolars extracted for orthodontic reasons from a 12 years old boy were obtained, and periodontal tissues were collected and cultured to obtain PDLCs. Cloned PDLCs were obtained by means of limited dilutions, and were characterized as follows: alkaline phosphatase activity, collagen III production and bone-like nodules formation. LPS and rhTNF-alpha were added into culture media and their effects on PDLC proliferation and OPG secretion were observed. The OPG concentrations in cell culture supernatants were detected by sandwich ELISA. Living cell numbers were demonstrated by MTT test. The average levels of OPG secretion by a single cell were calculated by dividing OPG concentration with MTT result.</p><p><b>RESULTS</b>rhTNF-alpha above 10 micro g/L decreased the mtt and opg detecting results, but increased the opg/mtt values (P < 0.05). However, LPS had no effect on mtt, opg or opg/mtt values. Neither it had any interaction with rhTNF-alpha (P > 0.05).</p><p><b>CONCLUSIONS</b>TNF-alpha prohibits the proliferation of PDLCs but enhances their OPG secretion. However, LPS has no effect on neither side. Our works support the hypothesis that there may be an inverse feedback regulation pattern of increasing periodontal OPG production against local bone resorption activity. PDLCs might not be the natural target cells of LPS' direct cytotoxic effect.</p>