Quantitative Analysis of Blended Gasoline Octane Number Using Raman Spectroscopy with Backward Interval Partial Least Squares Method / 分析化学

The feature of gasoline Raman spectra which were used to study the quantitative analysis of the research octane number (RON) were extracted for the first time using backward interval partial least squares (BiPLS). In the experiment, the sample set partitioning based on joint x-y distances (SPXY) method was used to divide the training set, the cross validation set and the test set. And the robust regression algorithm was used to remove the abnormal sample. The partial least squares model was established using feature selected by the BiPLS algorithm. Compared with the model without feature selection, it was shown that the backward interval partial least squares algorithm could reduce the input dimension by 50.00%, and the root mean square error of cross validation(RMSECV) by 18.92% and the root mean square error of prediction (RMSEP) by 13. 86%. The backward interval partial least squares algorithm can effectively extract the feature from gasoline Raman spectrum,reduce the model complexity, and improve the prediction accuracy of the model,and has great application prospect in the quantitative analysis of research octane number.

Therapeutic Role of Tangshenkang Granule () in Rat Model with Diabetic Nephropathy / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the renal protective effect of Tangshenkang Granule () in a rat model of diabetic nephropathy (DN).</p><p><b>METHODS</b>Forty male Sprague-Dawley rats were randomly divided into control, DN, Tangshenkang and benazepril groups. DN model was established in the rats of DN, Tangshenkang and benazepril groups. Tangshenkang Granule solution and benazepril hydrochloride solution were intragastrically administered daily to the rats in the Tangshenkang and benazepril groups for 8 weeks, respectively. Urinary albumin and creatinine were detected. The albumin/creatinine (ACR) was calculated in addition to 24 h urinary protein (24-h UPr), serum creatinine (Scr), blood urea nitrogen (BUN), total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), and creatinine clearance rate (Ccr). Right kidneys were harvested for pathological observation using periodic acid-silver methenamine-Masson staining. The average glomerular diameter (DG), average glomerular (AG) and mesangial areas (AM) were measured. The thickness of glomerular basement membrane (TGBM) was detected using transmission electron microscope.</p><p><b>RESULTS</b>Compared with rats in the control group, rats in the DN group showed significantly decreased body weight, increased hypertrophy index, 24-h urinary volume, 24-h UPr, ACR, Scr, BUN, Ccr, blood lipids as well as renal pathological indices including DG, AG, AM, AM/AG and TGBM (P <0.05). Compared with the DN group, the weights of rats in the Tangshenkang and benazepril groups were significantly increased, and the renal hypertrophy indices were significantly decreased (P <0.05). The 24-h urinary volumes, ACR, 24-h UPr, Scr, BUN, Ccr, LDL, DG, AG, AM and TGBM were obviously decreased (P <0.05). Compared with the benazepril group, the Tangshenkang group showed significantly decreased levels of ACR, 24-h UPr, AG and AM (P <0.05).</p><p><b>CONCLUSIONS</b>Tangshenkang Granule decreased the urinary protein, attenuated the high glomerular filtration rate and improved lipid metabolism in DN rats, and prevented further injury induced by diabetic nephropathy.</p>

Effect of Tanshinone ⅡA injection on intestinal mucosal tight junction protein in severe rat septic models / 中国中西医结合急救杂志

Objective To discuss the influence of Tanshinone ⅡA on the tight junction protein of intestinal mucosal epithelial cells in rat severe septic models. Methods Seventy-five Sprague-Dawley (SD) rats were randomly divided into sham operation group, model group and Tanshinone ⅡA injection high (20 mg/kg), medium (10 mg/kg) and low (5 mg/kg) dose groups, each group 15 rats. Sepsis rat models were established by cecal ligation and puncture (CLP) method, in sham operation group, only switched abdominal surgery was performed without CLP. In Tanshinone ⅡA injection groups, different doses of Tanshinone ⅡA were injected intraperitoneally after modeling for 10 minutes and 6 hours; in sham operation and model groups, equal volume of normal saline was injected intraperitoneally at the same times as above. After operation, 3 L/kg of normal saline was injected into the caudal vein in all rats for fluid resuscitation.Twelve hours after operation, the rats were killed, the abdominal lymph nodes, liver, spleen and kidney tissues were taken for bacterial culture and calculating the rate of bacterial translocation; under microscope, the histopathological changes of ileum mucosal tissues were examined and Chiu scoring was carried out; TdT-mediated dUTP nick end labeling (TUNEL) was applied to detect the ileum mucosal epithelial cell apoptosis and calculating the index (AI);fluorescence immunoassay and Western Blot methods were used to measure the contents and protein expression levels of tight junction protein, junctional adhesion molecule-1 (JAM), Claudin-1, Zonula occludens-1 (ZO-1), Occludin, c-Fos and Tryptase. Results ① In bacterial cultures of abdominal lymph node, liver, spleen and kidney, the positive rate of mesenteric lymph node was the highest, followed by liver and spleen, mainly Escherichia coli, Proteus mirabilis, etc. The highest positive rate of bacterial culture was in model group (38.8%), followed by low dose of Tanshinone ⅡA injection group (35.0%), and the lowest was 16.6% in high dose Tanshinone ⅡA injection group, the differences being statistically significant in comparisons between any pair of groups (all P < 0.05). ② Pathological examination showed that the pathological changes of ileum mucosa were obvious and the Chiu score (4.17±0.98 vs. 0) and AI (11.70±2.87 vs. 2.17±0.80) in model group were significantly higher than those in sham group (all P < 0.05); with the increase of dosage of Tanshinone ⅡA injection, the pathological changes of rat ileum mucosa were improved gradually, the Chiu score and AI were decreased gradually, and the degrees of decrease in high dose Tanshinone ⅡA group were more significant than those in model group (Chiu score: 1.12±0.79 vs. 4.17±0.98, AI: 3.65±1.98 vs. 11.70±2.87, both P < 0.05).③ Immunofluorescence staining showed that the positive staining of protein JAM, ZO-1 and c-Fos were all green in color, Claudin-1, Occludin and Tryptase were all red in color, the localizations of all of them were in the cytoplasm, the protein expression of JAM, Claudin-1, ZO-1, Occludin from strong to weak in turn were Sham group, high, medium, low dose Tanshinone ⅡA group and model group, the expression of c-Fos, Tryptase from strong to weak in turn were model group, low, medium, high dose Tanshinone ⅡA group and Sham group. ④ Western Blot showed that the expressions of ileum tissue JAM, Claudin-1, ZO-1 and Occludin in model group were all significantly lower than those of the sham group, while the expressions of c-Fos, Tryptase were obviously higher than those of the sham group, with the increase of dosage of Tanshinone ⅡA, the expressions of JAM, Claudin-1, ZO-1 and Occludin were increased gradually and the protein expressions of c-Fos and Tryptase were gradually decreased, and the changes in high dosage group of Tanshinone ⅡA were more significant than those in low and moderate groups [JAM (gray value): 25.39±1.82 vs. 12.41±1.34, 19.45±1.66, Claudin-1 (gray value): 28.44±1.56 vs.17.26±1.46, 21.23±1.34, ZO-1 (gray value): 28.84±1.59 vs. 16.45±1.21, 24.22±1.46, Occludin (gray value): 25.49±1.63 vs. 13.34±1.45, 19.45±1.37, c-Fos (gray value):15.76±1.36 vs. 27.84±1.36, 21.22±1.73, Tryptase (gray value): 14.44±1.41 vs. 28.14±1.38, 22.32±1.57], all the above comparisons of different dosage groups were statistically significant (all P < 0.05). Conclusion Tanshinone ⅡA injection may improve intestinal wall structure and reduce bacterial translocation by improving the intestinal mucosal tight junction protein in sepsis model rats, and this effect is positively correlated to Tanshinone ⅡA dosage.

An observation on interference mechanism of Shenfu injection on ghrelin in rats with severe sepsis / 中国中西医结合急救杂志

Objective To observe the effect of Shenfu injection on intestinal function in rats with sepsis. Methods Forty Sprague-Dawley (SD) rats were randomly divided into four groups: sham operation, sepsis model, low and high concentration Shenfu injection groups, each groupn = 10. The sepsis model was replicated by cecal ligation and puncture (CLP), while the rate in sham operation group just underwent abdominal incision without CLP. Ten minutes after CLP, the low and high dose Shenfu injection groups were given 5 mL/kg and 10 mL/kg Shenfu intravenous injection via a tail vein respectively. The rats in the model group were treated by intravenous injection of 10 mL/kg normal saline through a tail vein in 10 minutes after CLP. Twelve hours later, the rats were sacrificed. The levels of Ghrelin, Gastrin, tumor necrosis factor-α (TNF-α), high mobility group B1 protein (HMGB1), myeloperoxidase (MPO) and diamine oxidase (DAO) activity in serum were detected by enzyme linked immunosorbent assay (ELISA). The levels of protein of Ghrelin and gastrin receptor (GHSR) were detected by Western Blot. Under light microscope, the histopathological changes in intestinal mucosa were investigated, and Chiu score was determined, and the apoptosis index (AI) of intestinal mucosal epithelial cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Results Compared with sham operation group, in model group, the levels of Ghrelin and Gastrin in serum were significantly decreased [Ghrelin (ng/L): 121.23±3.53 vs. 146.28±5.43, Gastrin (ng/L): 81.78±3.27 vs 102.78±4.07], the serum levels of TNF-α and HMGB1 were markedly increased [TNF-α (mg/L): 93.71±3.66 vs. 11.69±1.44, HMGB1 (mg/L): 76.25±4.12 vs. 22.41±3.08], the DAO activity and protein expressions of Ghrelin and GHSR of intestinal tissue were obviously decreased [DAO (U/mL): 14.64 ±0.68 vs. 25.13±1.98, Ghrelin (grey value): 0.12±0.02 vs. 0.23±0.04, GHSR (grey value): 0.18±0.02 vs. 0.32±0.03], the MPO activity in intestinal tissue, Chiu score of intestinal mucosa and AI of ileum mucosal epithelial cells were remarkably increased [MPO (mg/L): 175.98±6.95 vs. 45.64±4.48, Chiu score: 3.90±0.52 vs. 0.30±0.30, AI: 29.31±1.65 vs. 5.45±1.35, allP 0.05). Under light microscope, the pathological changes were seen as follows: destruction and obvious edema of intestinal mucosal villi, ulcer formation, significant perivascular hemorrhage, presence of neutrophil infiltration and fracture of basement membrane in model group, while in low and high Shenfu groups, the intestinal villi had little defect, focal necrosis, small amounts of hemorrhage and neutrophil infiltration. Conclusions Shenfu injection can significantly improve the abnormal expressions of serum Ghrelin, reduce the levels of serum TNF-α and HMGB1, lowered MPO activity and enhance DAO activity in intestinal tissue, alleviate pathological changes in ileum mucosa, and decrease AI of ileum mucosal epithelial cells in rats with sepsis. And the degree of therapeutic effect is proportional to the Shenfu injection dose.

Effect of electro-acupuncture at zusanli (ST36) on the expression of ghrelin and HMGB1 in the small intestine of sepsis rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the expression of Ghrelin and high mobility group protein B1 (HMGB1) in the serum and the intestinal tissue of sepsis model rats, and to evaluate the effect of electro-acupuncture (EA) at Zusanli (ST36) on the expression of HMGB1 and Ghrelin.</p><p><b>METHODS</b>Forty-eight male Wistar rats were randomly divided into four groups, i.e., the sham-operation (sham), the cecal ligation and puncture group (CLP), the CLP + EA at Zusanli (ST36) group (EA), and the CLP + Ghrelin receptor blocking agent + EA group (GHSRA), 12 in each group. A sepsis rat model was prepared by CLP. The incision of the abdominal wall was immediately sutured along the ventral midline for rats in the Sham group. In the EA group EA at Zusanli (ST36) was performed 20 min after CLP surgery with the constant voltage (2 - 100 Hz, 2 mA) for 30 min. In the GHSRA group, Ghrelin receptor blocking agent, [D-Arg1, D-Phe5, D-Trp79, Leu11]-substance P (700 nmol/kg), was administered through intravenous injection immediately after CLP, and 20 min later, EA at Zusanli (ST36) was performed in the same way as for rats in the EA group. Blood samples were withdrawn 12 h after CLP. The serum levels of Ghrelin and HMGB1 were detected using ELISA. Ghrelin expressions and the number of Ghrelin immunopositive cell in the jejunum were determined by immunohistochemistry. HMGB1 contents of the jejunum tissue were detected by Western blotting.</p><p><b>RESULTS</b>Compared with the Sham group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased and levels of Ghrelin and the expression rate of immunopositive cells significantly decreased in the CLP group (P < 0.05). Compared with the CLP group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly decreased, but levels of Ghrelin and the expression rate of immunopositive cells significantly increased in the EA group (P < 0.05). Compared with the EA group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased in the GHSRA group (P < 0.05), but there was no statistical difference in levels of Ghrelin between the two groups (P > 0.05). The serum level of HMGB1 was negatively correlated with Ghrelin in the Sham group, the CLP group, and the EA group (r = -0. 528, P < 0.01).</p><p><b>CONCLUSIONS</b>EA at Zusanli (ST36) could inhibit the expression of HMGB1 in the jejunum of septic rats, and promote the expression of Ghrelin. The expression of HMGB1 was inhibited by Ghrelin receptor blocking agent, which suggested that the anti-inflammation of EA at Zusanli (ST36) might be associated with Ghrelin.</p>

Meiosis related gene expression in rat spermatogenesis / 中华男科学杂志

<p><b>OBJECTIVE</b>To find the expression of specific genes related to the meiosis of germ cells during spermatogenesis in the rat testis.</p><p><b>METHODS</b>Segments of seminiferous tubules were obtained from the adult male SD rats, at stages XIII - I of meiosis, and the interstitial cells of the same testis were isolated under the stereomicroscope. The total RNAs of stages XIII - I segments and the testicular interstitial cells were extracted respectively, and mRNA differential display RT-PCR (DDRT-PCR) was conducted. The obtained cDNA fractions were purified and recovered, the reverse dot blot hybridization, sub-clones and screens of blue/white dots performed, and the results of sub-clones were identified by restriction endonuclease EcoR I digestion.</p><p><b>RESULTS</b>Sixteen differential cDNA fractions were obtained through primary DDRT-PCR, 7 from stages XIII - I segments and 9 from the testicular interstitial cells. Another 11 were selected for further screening by reverse dot blot hybridization, their size ranging from 200 to 500 bp, of which 6 were from the stages XIII - I segments of seminiferous tubules and the other 5 from the rat testicular interstitial cells. All of the 11 cDNA fractions were sub-cloned and screened by blue/white dots.</p><p><b>CONCLUSION</b>Specifically expressed differential cDNA fractions can be obtained and primarily identified from testicular interstitial cells and the seminiferous tubules, which, as the sequence tags of the testicular meiotic expression, deserve further investigation.</p>

Specific expression of beta-actin during spermatogenesis in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To screen the stage-specific expression proteins during rats spermatogenesis, and to investigate the beta-actin expression and localization in the tissues of rat testicular.</p><p><b>METHODS</b>Highly enriched type A spermatogonia, pachytene spermatocytes and round spermatids were isolated by STAPUT method (sedimentation velocity at unit gravity, with 2% - 4% BSA gradient in DMEM/F12 medium) respectively to get the total proteins. The difference of protein expression between the three kinds of cells was analyzed by two-dimensional electrophoresis. Then the distribution of beta-actin in rat testicular tissues was investigated using specific anti-beta-actin antibodies by immunohistochemical method.</p><p><b>RESULTS</b>beta-actin was identified as a stage-specific expression protein by two-dimensional electrophoresis. beta-actin protein was more strongly expressed in type A spermatogonia and pachytene spermatocytes, but not in round spermatids. The immunohistochemical results showed that beta-actin was mainly located in the cytoplasm of type A spermatogonia and pachytene spermatocytes and in the nuclei of nearly mature spermatids.</p><p><b>CONCLUSION</b>beta-actin protein is a stage-specific expressed protein and may play an important role in spermatogenesis.</p>

Effect of experimental varicocele on structure and function of epididymis in adolescent rats / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To study the effect of experimental left varicocele (ELV) on epididymal structure and function in adolescent Sprague-Dawley rats.</p><p><b>METHODS</b>ELV was induced by partial ligation of the left renal vein. Sham-operated animals served as the controls. Four and 8 weeks after the operation, the histological, ultrastructural and biochemical (alpha-glucosidase activity and carnitine content) changes in different segments of the epididymis were observed.</p><p><b>RESULTS</b>In the treated animals, there were degeneration of the epididymal epithelium and edema of the interstitial tissue; numerous shedding cells, residual bodies, deformed sperm and macrophages appeared in the epididymal lumen. Morphometric measurement indicated a significant reduction in the epididymal tubular diameter (P<0.05) and a significant increase in the epididymal interstitial area (P<0.05) compared with the controls. Ultrastructural study showed sparse microvilli of the columnar epithelium, increased and enlarged lysosomes in the principal cells with defected organelles and the presence of large cytoplasmic vacuoles. The protein and carnitine contents and the alpha-glucosidase activity in the caput, corpus and cauda epididymis of the ELV rats were lower than those of the controls (P<0.05).</p><p><b>CONCLUSION</b>There were structural and functional changes in the epididymis of adolescent ELV rats, which may contribute to the infertility caused by varicocele.</p>