Quantification of Fat Concentration and Vascular Response in Brown and White Adipose Tissue of Rats by Spectral CT Imaging

Involvement of phosphatase and tensin homolog-induced putative kinase 1-Parkin-mediated mitophagy in septic acute kidney injury / 中华医学杂志(英文版)

BACKGROUND@#Studies have reported mitophagy activation in renal tubular epithelial cells (RTECs) in acute kidney injury (AKI). Phosphatase and tensin homolog-induced putative kinase 1 (PINK1) and E3 ubiquitin-protein ligase Parkin are involved in mitophagy regulation; however, little is known about the role of PINK1-Parkin mitophagy in septic AKI. Here we investigated whether the PINK1-Parkin mitophagy pathway is involved in septic AKI and its effects on cell apoptosis in vitro and on renal functions in vivo.@*METHODS@#Mitophagy-related gene expression was determined using Western blot assay in human RTEC cell line HK-2 stimulated with bacterial lipopolysaccharide (LPS) and in RTECs from septic AKI rats induced by cecal ligation and perforation (CLP). Autophagy-related ultrastructural features in rat RTECs were observed using electron microscopy. Gain- and loss-of-function approaches were performed to investigate the role of the PINK1-Parkin pathway in HK-2 cell mitophagy. Autophagy activators and inhibitors were used to assess the effects of mitophagy modulation on cell apoptosis in vitro and on renal functions in vivo.@*RESULTS@#LPS stimulation could significantly induce LC3-II and BECN-1 protein expression (LC3-II: 1.72 ± 0.05 vs. 1.00 ± 0.05, P < 0.05; BECN-1: 5.33 ± 0.57 vs. 1.00 ± 0.14, P < 0.05) at 4 h in vitro. Similarly, LC3-II, and BECN-1 protein levels were significantly increased and peaked at 2 h after CLP (LC3-II: 3.33 ± 0.12 vs. 1.03 ± 0.15, P < 0.05; BECN-1: 1.57 ± 0.26 vs. 1.02 ± 0.11, P < 0.05) in vivo compared with those after sham operation. Mitochondrial deformation and mitolysosome-mediated mitochondria clearance were observed in RTECs from septic rats. PINK1 knockdown significantly attenuated LC3-II protein expression (1.35 ± 0.21 vs. 2.38 ± 0.22, P < 0.05), whereas PINK1 overexpression markedly enhanced LC3-II protein expression (2.07 ± 0.21 vs. 1.29 ± 0.19, P < 0.05) compared with LPS-stimulated HK-2 cells. LPS-induced proapoptotic protein expression remained unchanged in autophagy activator-treated HK-2 cells and was significantly attenuated in PINK1-overexpressing cells, but was remarkably upregulated in autophagy inhibitor-treated and in PINK1-depleted cells. Consistent results were observed in flow cytometric apoptosis assay and in renal function indicators in rats.@*CONCLUSION@#PINK1-Parkin-mediated mitophagy might play a protective role in septic AKI, serving as a potential therapeutic target for septic AKI.

Involvement of phosphatase and tensin homolog-induced putative kinase 1–Parkin-mediated mitophagy in septic acute kidney injury / 中华医学杂志(英文版)

Background@#Studies have reported mitophagy activation in renal tubular epithelial cells (RTECs) in acute kidney injury (AKI). Phosphatase and tensin homolog-induced putative kinase 1 (PINK1) and E3 ubiquitin-protein ligase Parkin are involved in mitophagy regulation; however, little is known about the role of PINK1-Parkin mitophagy in septic AKI. Here we investigated whether the PINK1-Parkin mitophagy pathway is involved in septic AKI and its effects on cell apoptosis in vitro and on renal functions in vivo.@*Methods@#Mitophagy-related gene expression was determined using Western blot assay in human RTEC cell line HK-2 stimulated with bacterial lipopolysaccharide (LPS) and in RTECs from septic AKI rats induced by cecal ligation and perforation (CLP). Autophagy-related ultrastructural features in rat RTECs were observed using electron microscopy. Gain- and loss-of-function approaches were performed to investigate the role of the PINK1-Parkin pathway in HK-2 cell mitophagy. Autophagy activators and inhibitors were used to assess the effects of mitophagy modulation on cell apoptosis in vitro and on renal functions in vivo.@*Results@#LPS stimulation could significantly induce LC3-II and BECN-1 protein expression (LC3-II: 1.72 ± 0.05 vs. 1.00 ± 0.05, P < 0.05; BECN-1: 5.33 ± 0.57 vs. 1.00 ± 0.14, P < 0.05) at 4 h in vitro. Similarly, LC3-II, and BECN-1 protein levels were significantly increased and peaked at 2 h after CLP (LC3-II: 3.33 ± 0.12 vs. 1.03 ± 0.15, P < 0.05; BECN-1: 1.57 ± 0.26 vs. 1.02 ± 0.11, P < 0.05) in vivo compared with those after sham operation. Mitochondrial deformation and mitolysosome-mediated mitochondria clearance were observed in RTECs from septic rats. PINK1 knockdown significantly attenuated LC3-II protein expression (1.35 ± 0.21 vs. 2.38 ± 0.22, P < 0.05), whereas PINK1 overexpression markedly enhanced LC3-II protein expression (2.07 ± 0.21 vs. 1.29 ± 0.19, P < 0.05) compared with LPS-stimulated HK-2 cells. LPS-induced proapoptotic protein expression remained unchanged in autophagy activator-treated HK-2 cells and was significantly attenuated in PINK1-overexpressing cells, but was remarkably upregulated in autophagy inhibitor-treated and in PINK1-depleted cells. Consistent results were observed in flow cytometric apoptosis assay and in renal function indicators in rats.@*Conclusion@#PINK1-Parkin-mediated mitophagy might play a protective role in septic AKI, serving as a potential therapeutic target for septic AKI.

Serum epidemiology of five strong pathogenicity human adenoviruses natural infection in Tupaia belangeri / 中国人兽共患病学报

In order to explore the possibility of human adenovirus infection with tree shrews,the neutralizing antibody ti-ters of five kinds of human adenoviruses (HAdv)in the serum of tree shrews were analyzed.The levels of Ad3,Ad4,Ad7, Ad14 and Ad55 neutralizing antibody were detected by virus neutralization test.The results showed that the positive rate of four adenoviruses in group B were higher than Ad4 in group E,and the positive rates respectively were Ad14 (55.88%),Ad3 (47.06%),Ad55 (29.71%),Ad7 (14.71%)and Ad4 (8.82%).The antiserum mainly mixed with Ad3,Ad14 and Ad55 anti-body.Five species of human adenovirus can be naturally infected with tree shrews.Tree shrews are used as experimental ani-mals to establish human adenovirus infection model is alternative.

Effect of safflower yellow on atherosclerosis in rabbits with hyperlipidemia / 国际药学研究杂志

Objective To investigate the effect of safflower yellow injection on atherosclerosis in rabbits with hyperlipidemia.Methods Ninety-six New Zealand rabbits were randomly divided into four groups:the control group, model group, safflower yellowtest group (10.9, 5.45 and 2.725 mg/kg) and the positive control (atorvastatin) group. The control group was fed with normal feed, while the other three groups were fed with high fat diet for 8 weeks, combined with intraperitoneal injection of vitamin D3, to establish hyperlipidemia model. Then, the three-dosage safflower yellow-test groups were given intraperitoneal injection of safflower yellow (10.9, 5.45 and 2.725 mg/kg), respectively, the positive control group was given atorvastatin calcium[2 mg/ (kg·d) ]by intragastric administration, and the control and model groups were orally given an equal volume of normal saline, all once a day every day for 8weeks. After 16 h fasting following the last administration, the body weight, total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), oxidized low density lipoprotein (OX-LDL), matrix metalloproteinases 9 (MMP-9), tissue inhibitors of metalloproteinase 1 (TIMP-1), apolipoprotein E (ApoE), low density lipoprotein receptor (LDL-R), and scavenger receptor class B type1 (SR-B1) levels were measured. The morphological changes of thoracic aortas were examined by HE staining. Results At the 16 th week, compared with the control group, the body weight as well as the TC, TG, LDL-C, OX-LDL, MMP-9, HIF-1α, VEGF, VCAM-1 and PF4 level were all increased significantly (P＜0.05), while the level of HDL-C, ApoE, LDL-R, and SR-B1 decreased significantly (P＜0.05), accompanied with the atherosclerotic changes in the thoracic aortas indicated by the HE staining in the model group. Compared with the model group, the body weight as well as the TC, LDL-C, OX-LDL, MMP-9, HIF-1α, VEGF, VCAM-1 and PF4 level were decreased significantly (P＜0.05), and the TIMP-1 level increased (P＜0.05) in all of the three-dosage safflower yellow-test groups. Meanwhile, compared with the model group, in the 10.9 and 5.45 mg/kg safflower yellow groups, the TG level were decreased and the ApoE and SR-B1 levels were increased significantly (P＜0.05). On the other hand, the LDL-R level significantly increased only in the safflower yellow 10.9 mg/kg group (P＜0.05). HE staining showed a significant reduction in atherosclerorotic changes of the thoracic aorta in the safflower yellow-test groups. Conclusion Safflower yellow may inhibit the progression of atherosclerosis by regulating the lipid metabolism and MMP-9/TIMP-1 balance and also by inhibiting the HIF-1α, VEGF, VCAM-1 and PF4 expression.

Cardio-protective role of gingerol along with prominent anti-diabetic cardiomyopathy action in a streptozotocin-induced diabetes mellitus rat model

Objective: Diabetic cardiomyopathy [DCM] is characterized as a coronary heart disease which expands during diabetes due to alterations in the myocardial function and structure. The currentstudy intends to elucidate the protective effect of gingerol on DCM in a streptozotocin [STZ]-induced diabetes mellitus [DM] rat model

Materials and Methods: In this experimental study, the animals were divided into three groups: normal control, DM control, and DM+gingerol [10 mg/kg]. The body weights of all rats were estimated at regular intervals. The myocardial profile, oxidative stress, and activities of metabolic enzymes were also scrutinized. The proinflammatory cytokine levels together with cellular protein expression connected with apoptosis were estimated via Western blot analysis

Results: The rats that suffered from DCM exhibited abnormal levels of myocardial markers, aberrant metabolic enzymatic activity, elevated concentrations of inflammatory factors, and enhanced oxidative stress parameters along with increased cell death apoptosis. Whereas gingerol showed protective effects on the treated rats by an improved antioxidant defense system

Conclusion: The current findings suggested that gingerol is effective in the treatment of DCM by inhibition of inflammation and oxidative stress

Simultaneous determination of seven constituents in Artemisia argyi leaves by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of neochlorogenic acid,cryptochlorogenic acid,chlorogenic acid,caffeic acid,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C in the leaves of Artemisia argyi Levl.et Vant..METHODS The analysis of 50％ methanol extract of A.argyi leaves was performed on a 30 ℃ Prevail C1s column (4.6 mm ×250 mm,5 μmn),with the mobile phase comprising of acetonitrile-water flowing at 1.0 rnL/min in a gradient elution manner,and the detection wavelength was set at 325 nm.RESULTS Seven constituents showed good linear relationships within their own ranges (r ≥ 0.999 5),whose average recoveries were 98.28％-101.11％ with the RSDs of 1.04％-2.59％.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of A.argyi leaves.

Simultaneous determination of seven constituents in Artemisia argyi leaves by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of neochlorogenic acid,cryptochlorogenic acid,chlorogenic acid,caffeic acid,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C in the leaves of Artemisia argyi Levl.et Vant..METHODS The analysis of 50％ methanol extract of A.argyi leaves was performed on a 30 ℃ Prevail C1s column (4.6 mm ×250 mm,5 μmn),with the mobile phase comprising of acetonitrile-water flowing at 1.0 rnL/min in a gradient elution manner,and the detection wavelength was set at 325 nm.RESULTS Seven constituents showed good linear relationships within their own ranges (r ≥ 0.999 5),whose average recoveries were 98.28％-101.11％ with the RSDs of 1.04％-2.59％.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of A.argyi leaves.

Correlation of hyperhomocysteinemia and cognitive impairment in patients with cerebral infartion combined chronic obstructive pulmonary disease and drug treatment / 中国生化药物杂志

Objective To investigate the relationship between serum homocysteine ( Hcy ) and cognitive impairment in patients with cerebral infarction combined chronic obstructive pulmonary disease (COPD) after stroke, and observe the plasma Hcy levels and cognitive function improvement when treated with folic acid and vitamin B12 .Methods 87 acute cerebral infarction combined with COPD patients as the research object, then the general clinical data were recorded, hematology indexes ( Hcy, folic acid, vitamin B12 ) were determined, and their cognitive function with a simple mental state scale (MMSE) was assayed.According to the plasma Hcy levels, the subjects were divided into Hcy-normal group (n =21) and Hcy-increased group (n=66), then compare the cognitive function between the two groups.Hcy-increased subjects were randomly divided into intervention group (conventional treatment +folic acid 2.5 mg +VitB12 0.5 mg) and control group (conventional treatment).After six months follow-up, we retested plasma Hcy levels and MMSE assessment, comparison of plasma Hcy concentration change and cognitive function improvement between two groups.Results Compared with Hcy-normal group, plasma folic acid, VitB12 levels significantly decreased in Hcy-increased group (P<0.05).And Hcy concentration was negatively correlated with folic acid(r=-0.351,P =0.000)and VitB12(r=-0.242,P=0.015)levels.In addition, the MMSE, directional force and delayed recall score decreased in Hcy-increased group compared with the Hcy-normal group ( P<0.05 ).Hcy levels were significantly lower than the baseline level (P<0.05), MMSE and the sub-project of cognitive function score increased after treated with folic acid and VitB12 for six months, although there was no statistically significant difference.Conclusion Plasma Hcy level is associated with cognitive impairment in patients with cerebral infarction combined chronic obstructive pulmonary disease (COPD), patients treated with folic acid and VitB12 may help slow the recent cognitive dysfunction after stroke in the near future.

Ursolic acid inhibits T-cell activation through modulating nuclear factor-κ B signaling / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of ursolic acid (UA) on T-cell proliferation and activation, as well as to examine its effect on nuclear factor-κB (NF-κB) signaling pathway in T cells.</p><p><b>METHODS</b>T-cells isolated from BALB/c mice were incubated with UA at concentrations ranging from 5-30 μmol/L in the presence of phorbol 12-myristate 13-acetate (PMA) or PMA plus ionomycin. The proliferation of T cells was measured by the MTT assay. The expressions of CD69, CD25, and CD71 on T-cell surface were analyzed using flow cytometry. The level of interleukin-2 (IL-2) in the culture supernatant of activated T cells was quantified by enzyme-linked immunosorbent assay (ELISA). The level of phosphorylated IκB-α (p-IκB-α) in total protein and p65, a subunit of NF-κB, nuclear translocation were measured by Western blot analysis.</p><p><b>RESULTS</b>UA in a dose-dependent manner significantly decreased the proliferation and inhibited the surface expressions of CD69, CD25, and CD71 in murine T lymphocytes upon in vitro activation (P<0.01). Significant reduction of IL-2 production was found in activated T cells treated with UA (P<0.01). The PMA-induced increase in p-IκB-α protein was inhibited, and nuclear translocation of p65 from the cytoplasm was blocked by UA.</p><p><b>CONCLUSION</b>UA is a potent inhibitor for T cell activation and proliferation; these effects are associated with the inhibition of NF-κB signaling pathway.</p>

Analysis of human brucellosis surveillance in Weinan city of Shaanxi province in 2009 / 中国地方病学杂志

Objective To survey and analyze characteristics of brucellosis epidemic in Weinan city of Shaanxi province for the purpose of setting up prevention and control measures for the disease. Methods According to "The Executing Plan for the Work of Surveying Brucellosis Disease in Shaanxi Province", 35 villages(towns) of designated monitoring locations and 24 villages (towns) of randomized monitoring locations in five countries of Weinan were chosen to survey brucellosis disease. The five countries were Chengcheng, Dali, Heyang, Tongguan and Hancheng. High risk populations with a history of contacting livestock and livestock products aged between 7 and 60 underwent clinical and serology examination[rose bengal plate agglutination test(RBPT) and standard tube agglutination test(SAT)]. All manipulation methods and judging standards were in accord with the "Diagnostic Standard for Brucellosis" (WS 269-2007). Results In the designated monitoring location, a total of 8664 people at high risk were investigated, among whom 1407 people were tested by RBPT test and 27 people were positive,the positive rate was 1.92%(27/1407); 27 people were tested by SAT test and 27 people were positive, the positive rate was 100% (27/27); 25 people were diagnosed and the diagnosis rate was 92.59%(25/27). In the randomized monitoring location, a total of 3464 people at high risk were investigated, among whom 411 people were tested by RBPT test and 3 people were positive, the positive rate was 0.73%(3/411 ), 3 people were tested by SAT test which were all positive and made a definite diagnosis. Twenty-eight new cases were made a definite diagnosis and its incidence was 2.06 in a hundred thousand(28/1 361 618). Conclusions The infection of human brucellosis in Weinan city stays at higher level. The governments should increase input for the monitoring,investigating and disinfecting to prevent the disease from increasing and outspreading.

Effects of electroacupuncture at "Zusanli" (ST 36) and "Neiguan" (PC 6) on expression of GAP-43 in cerebral infarction rats / 中国针灸

<p><b>OBJECTIVE</b>To observe the effect of electroacupuncture on the neural plasticity in rats with cerebral infarction and investigate its mechanism.</p><p><b>METHODS</b>Sixty rats were randomly divided into a sham operation group, an acupuncture group and a non-acupuncture group, and each group was randomly divided into a 1-day subgroup, a 7-day subgroup and a 14-day subgroup. Cerebral infarction model was induced by the thread embolism method. The rats in the acupuncture group were treated by electroacupuncture at "Zusanli" (ST 36) and "Neiguan" (PC 6) for 20 minutes, once each day, while the rats in the sham operation group and the non-acupuncture group were just bound and fixed at the same time without acupuncture treatment. Neurological defects were assessed by Neurological Severity Score (NSS) and the changes of the expression of growth associated protein 43 (GAP-43) in peripheral nerve around cerebral infarction area were detected by immunohistochemistry technique.</p><p><b>RESULTS</b>The expression of the positive cells of GAP-43 around cerebral infarction area showed no significant distinction among the three groups at 1st day (P > 0.05), while the GAP-43 expression level of around cerebral infarction area in the acupuncture group (IOD:8. 990 1 +/- 0.098 7, 5.816 1 +/- 0.204 6) were significant higher than those in the sham operation group (IOD: 1.300 2 +/- 0.093 3, 1.362 6 +/- 0.216 6) and in the non-acupuncture group (IOD: 2.753 4 +/- 0.0875, 1.616 5 +/- 0.186 8) at 7th day and 14th day, respectively (all P < 0.01).</p><p><b>CONCLUSION</b>Electroacupuncture at "Zusanli" (ST 36) and "Neiguan" (PC 6) can improve the neural function and promote its remodeling in rats with cerebral infarction and the relevant mechanisms may be involved in enhancement of GAP-43 expressions around ischemic region.</p>

Microarray data analysis of genes related with erectile dysfunction in diabetic rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the changes of gene expression profiles associated with erectile dysfunction in diabetic rats.</p><p><b>METHODS</b>Affymetrix Gene Chip arrays from the Gene Expression Omnibus (GEO) were used to examine the alterations in the gene expression profiles between streptozotocin-induced diabetic rats and littermate controls, and the data were analyzed with GeneSifter microarray analysis software.</p><p><b>RESULTS</b>A total of 661 differentially expressed genes were identified, including 280 up-regulated and 381 down-regulated ones. Among the differentially expressed genes, kruppel-like factor 5 (klf5) was upregulated by 4.01 folds and ceruloplasmin(cp) by 5.14 folds; collagen, type XI, alpha1 was down-regulated by 5.84 folds and collagen, type I, alpha1 by 5.77 folds. The 661 differentially expressed genes involved such functional processes as glycoprotein biosynthesis, collagen fibril organization, angiogenesis in wound healing, triglyceride metabolism, cell proliferation and other important biological processes, and some pathways also involved such as fatty acid metabolism, neurodegenerative disorders, and ECM-receptor interactions.</p><p><b>CONCLUSION</b>Some genes such as klf5, cp, and collagen play important roles in the pathophysiology of diabetes-induced erectile dysfunction. Bioinformatic approaches offer a new means for identifying candidate genes and pathways relevant to the pathophysiology of diabetes-induced erectile dysfunction, highlighting also the potential complexity of this disorder.</p>

Calcitonin gene-related peptide induces phenotypic transformation of corpus cavernosum smooth muscle cells in diabetic rats with erectile dysfunction / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effect of the calcitonin gene related peptide (CGRP) on the phenotypic transformation of corpus cavernosum smooth muscle cells (CCSM) in diabetic rats with erectile dysfunction (ED).</p><p><b>METHODS</b>Models of diabetes and diabetic ED were established in male Sprague-Dawley rats by administration of streptozotocin, and CCSMs were primarily cultured and subjected to immunocytochemical assay. The cells were divided into a diabetic ED and a normal control group, and exposed to 0, 10, 60 and 100 nmol/L of CGRP for 24 hours. Then the relative expressions of calponin 1 (Cnn1) and osteopontin (OPN) mRNA were determined by real-time fluorescence quantitative RT-PCR (qRT-PCR).</p><p><b>RESULTS</b>The rate of SMalpha-actin positive cells in the CCSMs was (95.94 +/- 0.03) %. The expression of Cnn1 mRNA was significantly lower while that of OPN mRNA remarkably higher in the diabetic ED rats (4.41 +/- 0.29 and 5.28 +/- 0.32) than in the normal controls (10.35 +/- 0.62 and 1.32 +/- 0.24) (P < 0.01). Exposure to 100 nmol/L of CGRP significantly upregulated the expression of Cnn1 mRNA and downregulated that of OPN mRNA as compared with the unexposed rats (6.9 +/- 0.22 vs 4.41 +/- 0.29 and 3.26 +/- 0.31 vs 5.28 +/- 0.32, P < 0.01).</p><p><b>CONCLUSION</b>CGRP can transform the phenotype of CCSMs in diabetic ED rats from contractile to synthetic type.</p>

Proteomics study on the essence of wind syndrome caused by gan-yang hyperactivity in Chinese medicine / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To preliminarily study the essence of wind syndrome caused by Gan-yang hyperactivity (WSGH) in Chinese medicine at the protein expression level.</p><p><b>METHODS</b>WSGH was strictly differentiated from wind stirring due to yin deficiency syndrome in patients with intracerebral hemorrhage (ICH) and those with cerebral infarction (CI); from Gan-yang hyperactivity syndrome in patients with cervical spondylosis (CS); from wind syndrome induced by blood deficiency in patients with Parkinson's disease (PD) according to Chinese medicine syndrome typing standard. Control studies were performed. Peripheral blood mononuclear cells (PBMCs) were isolated from all patients of the aforesaid syndromes and healthy subjects. The total proteins were extracted, two-dimensional gel electrophoresis (2-DE) conducted and analyzed by PDQuest software. The peptide mass fingerprint (PMF) was determined using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). The SwissProt database was inquired using Mascot reference system. Proteins of different and same expressions in PBMCs of patients suffering from the same disease of different syndromes, different diseases of the same syndrome, and syndromes of the same kind were compared.</p><p><b>RESULTS</b>The 2-DE map of PBMCs' total proteins in the aforesaid syndrome groups and healthy subjects was established. Through comparison, analysis, and appraisement, there was 1 protein dot of the same expression and 22 protein dots of different expressions between ICH patients of WSGH and ICH patients of wind stirring due to yin deficiency syndrome. There were 6 protein dots of the same expression and 21 protein dots of different expressions between CI patients of WSGH and CI patients of wind stirring due to yin deficiency syndrome. There were 3 protein dots of the same expression and 12 protein dots of different expressions between CS patients of WSGH and CS patients of Gan-yang hyperactivity syndrome. There was no protein dot of the same expression and 12 protein dots of different expressions between PD patients of WSGH and PD patients of wind syndrome induced by blood deficiency. There were 13 protein dots of the same expression in different diseases of the same syndrome. There was 1 protein dot (Thioredoxin-dependent peroxide reductase, TPx) of the same expression in the four diseases of the same kind syndrome.</p><p><b>CONCLUSIONS</b>Different connotations of the essence existed (having multiple different protein expressions) in patients with the same disease of different syndromes. Syndromes of the same kind share the same material bases (having the same protein expression). These suggested that Chinese medicine syndrome has its own material bases and essence findable.</p>

Role of miR-145 in erectile dysfunction in diabetic rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of miR-145 in the corpus cavernosum smooth muscle tissue in the pathogenesis of erectile dysfunction (ED) in diabetic rats.</p><p><b>METHODS</b>The total RNA was extracted from the corpus cavernosum of a diabetic rat model with ED, diabetic rats with normal erectile function and normal rats, and the expression levels of miR145 were compared between the groups.</p><p><b>RESULTS</b>The expression of miR-145 was decreased in the corpus cavernosum of diabetic rats with ED.</p><p><b>CONCLUSION</b>Diabetes mellitus can cause ED in rats, in which process decreased expression of miR145 in the corpus cavernosum may play a role.</p>

A modified tissue culture method for culturing corpus cavernosum smooth muscle cells from the penis of Sprague-Dawley rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To culture rat corpus cavernosum smooth muscle cells in vitro using a modified tissue culture method.</p><p><b>METHODS</b>Fifteen male rats were randomized into 3 equal groups, namely enzyme digestion group, tissue culture group, and modified tissue culture group. The penis of the rats was separated carefully and cut into small pieces, and seeded onto culture flasks and cultured in complete medium consisting of DMEM containing 20% fetal calf serum at 37 degrees C; in a humidified atmosphere with 5% carbon dioxide. The cells growth was observed under phase contrast microscope and the smooth muscle cell specific proteins alpha-SM-actin and desmin were identified immunohistochemically.</p><p><b>RESULTS</b>The alpha-SM-actin-positive cell rate was 96.3% in rat corpus cavernosum smooth muscle and 23.8% in the fibroblasts, and the corpus cavernosum smooth muscle contained 74.4% desmin-positive cells while the fibroblasts showed no desmin positivity. Significant difference was found in the positive cell rate for desmin among the 3 groups, with the highest positive cell rate occurred in modified tissue culture group.</p><p><b>CONCLUSION</b>Desmin may serve as a marker for identifying corpus cavernosum smooth muscle cells. The modified tissue culture method can result in highly purified corpus cavernosum smooth muscle cells with intact structure and functions.</p>

Effect of Bizhongxiao decoction on proteomics of peripheral blood mononuclear cells in patients with rheumatoid arthritis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To probe in the possible acting mechanism of Bizhongxiao Decoction (BZXD) for treatment of early active rheumatoid arthritis (RA) by way of observing the two-dimensional gel electrophoresis map of proteins in peripheral blood mononuclear cells (PBMCs) of healthy persons and RA patients (intervened or un-intervened with BZXD), analyzing the differential proteins and seeking out the RA associated proteins.</p><p><b>METHODS</b>Eighteen patients with early active RA were randomized into the BZXD group and the methotrexate (MTX) group, nine in each group, they were treated with BZXD (contained 15 Chinese herbs, as Herba Hedyotis diffusae, Herba Sarcandrae glabrae, Radix Salviae miltiorrhizae, Caulis Trachelosperi, Rhizoma Drynariae, Semen Coicis, etc.) and MTX combined with nimesulide Tablets respectively, three months as a treatment course, and their blood samples were collected for observation. Besides, blood samples from 9 healthy persons were taken as normal controls. PBMCs were isolated from blood using lymphozytes separation medium, and total protein in the cells was extracted through immobilized pH gradient two-dimensional gel electrophoresis. After Coomassie brilliant blue G250 staining, gel-image analysis was performed using PDQuest software. The differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). Then partial proteins were validated by reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The 2-DE protein profile of PBMCs from healthy persons and RA patients before and 3 months after treatment were obtained, and 23 differential protein spots were found, 14 from 18 differential protein spots were successfully identified, of which 8 proteins were up-regulated and 6 proteins were down-regulated in RA patients as compared with control. After 3-month treatment, 5 differentially expressed proteins showed more obvious in the BZXD group than in the MTX group. RT-PCR verified that the expression of ApoA-I in all the three groups was consistent with the outcomes of 2-DE.</p><p><b>CONCLUSIONS</b>Some differentially expressed proteins exist in the PBMCs of RA patients, which may play a potential role in the pathogenesis of RA; BZXD may treat RA by way of regulating the expression of some differential proteins in patients.</p>

Establishment of a cell culture model of corpus cavernosum smooth muscle cells derived from diabetic rats with erectile dysfunction / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the method for culturing corpus cavernosum smooth muscle cells (CCSMs) derived from diabetic rats with erectile dysfunction (ED) for the study of ED caused by diabetes.</p><p><b>METHODS</b>CCSMs were isolated from the corpus cavernosum of diabetic rats with ED and cultured using a modified method of adherent tissue culture. The cultured cells were identified by immunohistochemistry and the cell morphology and proliferation were observed.</p><p><b>RESULTS</b>The primary culture of CCSM was performed successfully, and the cells were seen to migrate from the small tissue pieces 3 days later, reaching nearly confluence in 16-18 days. A typical "hill-valley" growth pattern was noted in the cell passaging. Immunohistochemical staining for alpha-smooth muscle actin (alpha-SM-actin) and desmin yielded positive results in the cells.</p><p><b>CONCLUSION</b>The modified method for adherent tissue culture is convenient and reliable in establishing the in vitro cell culture model of CCSMs from diabetic rats with ED, and the cultured CCSMs display a faster proliferation than normal CCSMs. No obvious differences in the cell morphology can be found between diabetic and normal CCSMs under light microscope.</p>

Retroperitoneal laparoscopic surgery combined with ureteroscopic lithotomy for treatment of renal and ureteral calculi / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the feasibility of retroperitoneal laparoscopic surgery combined with ureteroscopic lithotomy through the pelvis for treatment of renal and ureteral calculi.</p><p><b>METHODS</b>In February 2010, 2 patients with renal and ureteral calculi underwent retroperitoneal laparoscopic surgery combined with ureteroscopic lithotomy through the pelvis.</p><p><b>RESULTS</b>The operation time in these two cases was 70 and 80 min, and the volume of intraoperative blood loss was about 20 ml. The exposure was excellent, and the patient recovered rapidly without complications or residual calculi.</p><p><b>CONCLUSION</b>Retroperitoneal laparoscopic surgery combined with ureteroscopic lithotomy through the pelvis is feasible for treatment of renal and ureteral calculi.</p>