Effects of timosaponin B-II on differentiation of rat neural stem cells into tyrosine hydroxylase posi-tive neurons in vitro / 中华行为医学与脑科学杂志

Objective To explore the effect of timosaponin B-II ( TB-II) on the differentiation of neural stem cells (NSCs) into tyrosine hydroxylase (TH) positive neurons in neonatal rats. Methods The biological functions of self-proliferation and multi-differentiation of NSCs were identified by primary culture, cell proliferation counting,morphological observation and immunology. NSCs of SD rats were cultured in vitro and treated with different concentrations of TB-II (10 μg/ml,30 μg/ml ,100 μg/ml) for 7 days. Immuno-histochemistry was used to detect the effect of TB-II on the differentiation of NSCs into TH-positive neurons, and Western blot was used to detect the expression of TH protein in neurons. Results ( 1) The cultured cells had the ability to self-proliferation,expressed nestin protein and differentiated into neurons and glial cells. So the cultured cells were conformed to the biological function of neural stem cells. (2)Compared with the control group,the TH positive cell ratio of TB-II 30 μg/ml group and TB-II 100 μg/ml group increased ((10. 03± 1. 36)%),( 20. 01± 3. 37)%),(31. 32± 3. 98)%) ,the difference was significant ( t=6. 15, 16. 54,both P<0. 05). There was no significant difference between TB-II 10 μg/ml group and control group (P>0. 05). (3)Western results showed that the relative expression of TH protein in TB-II 30 g/ml group and TB-II 100 μg/ml group was higher than that in control group,the difference was statistically significant (con-trol group: (1. 02±0. 24),TB-II 30μg/ml group: (3. 64±1. 78),TB-II 100 μg/ml group: (5. 88±2. 34);t=12. 58,9. 15,both P<0. 05). There was no significant difference between TB-II 10 μg/ml group and con-trol group (P>0. 05). Conclusion TB-II can promote the differentiation of NSCs into TH-positive neurons.

Clinical evaluation of endoscopic anti-reflux mucosectomy for gastroesophageal reflux disease / 中华消化内镜杂志

Objective@#To assess the efficacy and safety of the endoscopic anti-reflux mucosectomy for gastroesophageal reflux disease.@*Methods@#Data of 18 patients with gastroesophageal reflux disease who underwent endoscopic anti-reflux mucosectomy at the First Affiliated Hospital of ZhengZhou University from December 2015 to July 2018 were retrospectively studied. The therapeutic effects (improvement of heartburn and reflux symptoms, 24 h esophageal pH monitoring) and complications were analyzed.@*Results@#Anti-reflux mucosectomy was performed successfully in all patients with successful rate of 100%. ESD was performed in 8 cases and EMR in 10 cases.24 h esophageal pH monitoring results showed that the Demeester score, the time percentage of pH < 4, total reflux events and reflux times of pH < 4 with time longer than 5 minutes after treatment were significantly lower than those before treatment (20.16±9.12 VS 74.16±20.03, (2.70±0.88)% VS (6.42±1.37)%, 43.78±19.68 VS 156.56±41.22, 2.89±1.68 VS 9.89±2.95, all P<0.05). No bleeding, perforation or infection was observed after the procedure. During a follow-up period of 3-34 months, symptom relief rate was 89% (16/18). A tightened cardiac was noted in 18 cases and recovery of mucosal damage was found in 16 patients.@*Conclusion@#Anti-reflux mucosectomy is safe, effective and easy to operate for gastroesophageal reflux disease.

Clinical evaluation of endoscopic anti-reflux mucosectomy for gastroesophageal reflux disease / 中华消化内镜杂志

Objective To assess the efficacy and safety of the endoscopic anti-reflux mucosectomy for gastroesophageal reflux disease. Methods Data of 18 patients with gastroesophageal reflux disease who underwent endoscopic anti-reflux mucosectomy at the First Affiliated Hospital of ZhengZhou University from December 2015 to July 2018 were retrospectively studied. The therapeutic effects ( improvement of heartburn and reflux symptoms, 24 h esophageal pH monitoring) and complications were analyzed. Results Anti-reflux mucosectomy was performed successfully in all patients with successful rate of 100％. ESD was performed in 8 cases and EMR in 10 cases. 24 h esophageal pH monitoring results showed that the Demeester score, the time percentage of pH < 4, total reflux events and reflux times of pH < 4 with time longer than 5 minutes after treatment were significantly lower than those before treatment (20. 16±9. 12 VS 74. 16±20. 03, (2. 70±0. 88)％ VS (6. 42±1. 37)％, 43. 78±19. 68 VS 156. 56±41. 22, 2. 89±1. 68 VS 9. 89±2. 95, all P<0. 05) . No bleeding, perforation or infection was observed after the procedure. During a follow-up period of 3-34 months, symptom relief rate was 89％ (16/18). A tightened cardiac was noted in 18 cases and recovery of mucosal damage was found in 16 patients. Conclusion Anti-reflux mucosectomy is safe, effective and easy to operate for gastroesophageal reflux disease.

Long-term efficacy of peroral endoscopic circular myotomy and full-thickness myotomy on treatment of achalasia of cardia / 中华消化内镜杂志

Objective To compare the long-term efficacy and complications of peroral endoscopic circular myotomy and full-thickness myotomy for patients with achalasia of cardia. Methods A retrospective analysis was performed on the data of 53 patients with achalasia of cardia, who underwent peroral endoscopic myotomy in the First Affiliated Hospital of Zhengzhou University from June 2012 to December 2014 and were followed-up regularly.Twenty-one patients underwent circular myotomy, and the other 32 patients underwent full-thickness myotomy. The postoperative long-term efficacy and gastroesophageal reflux complications of the two groups were compared. Results The effective rate of the circular myotomy group and the full-thickness myotomy group was 90. 5%( 19/21) and 100. 0%( 32/32), respectively ( P＝ 0. 152). There were no significant differences between the two groups on postoperative Eckardt scores, lower esophageal sphincter pressure and 4 s integrated relaxation pressure ( P ＞ 0. 05 ). The incidence of clinically relevant gastroesophageal reflux of full-thickness myotomy group was higher than that of circular myotomy group (40. 6% VS 14. 3%, χ2＝4. 174, P＝0. 041). Conclusion The long-term efficacy of circular myotomy is similar to that of full-thickness myotomy, but the incidence of clinically relevant gastroesophageal reflux is higher in full-thickness myotomy.

Analysis of cell morphology and immunophenotypic characteristics in 47 cases of multiple myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was to investigate the cell morphology and cell immune phenotypic characteristics in patients with multiple myeloma (MM).</p><p><b>METHODS</b>The flow cytometry with multiparametric direct immunofluorescence technique, and CD45/SSC and CD38(+)(+)/CD138(+) gating were used to measure cell markers CD138, CD38, CD56, CD117, CD3, CD13, CD33, CD19, CD7, CD20, CD22, CD34, CD28 in 47 MM patients. At the same time the morphology examination of bone marrow cells was performed.</p><p><b>RESULTS</b>The suspicious myeloma cell ratio in MM patients was 9.42%-74.25% detected by flow cytometry, moreover, the myeloma cell ratio detected by morphology examination was 11.0%-80.6%, there was a good correlation between the two detection methods (r(2) = 0.54, P < 0.001). The ratio of antigen positive expression was as follows: 74.46% for CD138, 100% for CD38, 57.44% for CD56, 40.42% for CD117, 6.38% for CD13, 19.15% for CD33, 8.51% for CD20, 27.66% for CD28, 2.12% for CD22, 4.25% for CD34, 0% for CD3, 0% for CD19, 0% for CD7.</p><p><b>CONCLUSIONS</b>CD45/SSC and CD38(+)/CD138(+) gating technique can accurately gate multiple myeloma cell sets which need analysis, the majority of myeloma cells expreses CD138, CD38, CD56 antigens. The immunophenotypic analysis combined with the cell morphology examination more contribute to the diagnosis and differential diagnosis of multiple myeloma.</p>

Phenotypic modulation of vascular smooth muscle cells in diabetes mellitus and intervention of traditional Chinese medicines / 中国中药杂志

Proliferation and migration of vascular smooth muscle cells (VSMC) are common pathological features of diabetic vascular complications,such as atherosclerosis and hypertension. Phenotypic modulation of VSMC is the basis for VSMC proliferation and migration. Therefore, studies on VSMC phenotypic modulation and its mechanisms in diabetes mellitus were of important significance to the prevention and therapy of diabetic vascular complications. This paper introduces VSMC phenotypic modulation and the underlying mechanisms in diabetes mellitus, and summarizes advance of studies on traditional Chinese medicine intervention upon VSMC phenotypic modulation, so as to provide reference for preventing and treating diabetic vascular complications with traditional Chinese medicines.

Metabolic shift of Corynebacterium acetoacidophilum-deltaldh under oxygen deprivation conditions / 生物工程学报

Lactate and succinate were produced by Corynebacterium acetoacidophilum from glucose under oxygen deprivation conditions. To construct knockout mutant, lactate dehydrogenase gene (ldh) of C. acetoacidophilum was deleted by double-crossover chromosome replacement with sacB gene. Comparing with the wild strain ATCC13870, ldhA-deficent mutant produced no lactate with glucose consumption rate decreased by 29.3%, while succinate and acetate concentrations were increased by 45.6% and 182%, respectively. Moreover, the NADH/NAD+ rate was less than 1 (about 0.7), and the activities of phosphoenolpyruvate carboxylase and acetate kinase of the ldhA-deficent mutant were enhanced by 84% and 12 times, respectively. Our studies show that succinicate and acetate production pathways are strengthened by blocking lactate synthesis. It also suggests that improving NADH supply and eliminating acetate generation are alternative strategies to get high succinate-producer.

A microarray analysis of angiogenesis modulation effect of Xuefu Zhuyu Decoction on endothelial cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the angiogenesis modulation mechanism of Xuefu Zhuyu Decoction () on the endothelial cell line ECV304.</p><p><b>METHODS</b>ECV304 cells were treated with 2.5% Xuefu Zhuyu Decoction-containing serum (XFZYD-CS) for 24 h, 48 h or 72 h. Thiazolyl blue tetrazolium bromide (MTT), fluorescence activating cell sorter (FACS), migration, adhesion and in vitro tube formation assays were conducted to confirm an angiogenesis effect of XFZYD at 3 time points. An analysis of angiogenesis regulator profiles was performed at 3 times with real-time polymerase chain reaction (RT-PCR) superarray.</p><p><b>RESULTS</b>At 48 h, XFZYD-CS induced ECV304 significantly improved cell viability, number in S phase, migration, adhesion and tube formation. At 24 h and 72 h, only cell migration was elevated. Microarray results showed that the expression of 27 angiogenesis-related genes was changed.</p><p><b>CONCLUSION</b>XFZYD-CS treatment induced angiogenesis on ECV304 cells with significant cellcular changes occurring at 48 h and genetic changes as early as 24 h.</p>

Study on function of VEGF pathway in promoting angiogenesis with Xuefu Zhuyu Tang / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the function of VEGF pathway in promoting angiogenesis with Xuefu Zhuyu Tang (XFZYT).</p><p><b>METHOD</b>Serum pharmacology technique was adopted to treat endothelial cells ECV304 with XFZYD containing serum and blank serum with concentrations of 1.25%, 2.5% and 5%. Methyl thiazolyl tetrazolium (MTT) assay, boyden chamber migration assay and in vitro vessel formation assay were used to test endothelial cell proliferation, migration and angiogenesis capacities. ELISA, immunohistochemistry and RT-PCR were used to detect secretion and expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR2).</p><p><b>RESULT</b>XFZYT-CS with a concentration of 1.25% only affected angiogenesis capacity in vitro. XFZYT-CS with a concentration of 2. 5% promoted cell proliferation, migration and angiogenesis capacities and significantly increased VEGF level and VEGF/VEGFR2 expressions. XFZYT-CS with a concentration of 5% only up-regulated intracellular VEGF expression and thereby improving endothelial cell proliferation, migration and angiogenesis.</p><p><b>CONCLUSION</b>XFZYT can induce endothelial cell proliferation, migration and angiogenesis by up-regulating VEGF-VEGFR2 pathway, which partially proves its angiogenesis effects.</p>

Experimental study on the suppression of sodium nitroprussiate-induced chondrocyte apoptosis by Tougu Xiaotong Capsule (透骨消痛胶囊)-containing serum / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the mechanism of action of Tougu Xiaotong Capsule (透骨消痛胶囊, TGXTC) ex vivo in suppressing chondrocyte (CD) apoptosis induced by sodium nitroprussiate (SNP).</p><p><b>METHODS</b>Thirty New Zealand rabbits, 2 months old, were randomized by lottery into five groups, six in each: the blank group treated with saline, the positive control group treated with Zhuanggu Guanjie Pill (壮骨关节丸, 70 mg/kg), and the three experimental groups, EGA, EGB, and EGC, treated with low dose (35 mg/kg), moderate dose (70 mg/kg), and high dose (140 mg/kg) of TGXTC, respectively. All treatments were administered via gastrogavage twice a day for 3 days. Arterial blood was collected from the abdominal aorta and drug or drug metabolites-containing serum was prepared. CDs obtained from knee joints of 16 four-week-old New Zealand rabbits were cultured to the third passage and confirmed by toluidine blue staining. SNP of various final concentrations (0, 0.5, 1.0, and 2.0 mmol/L) was used to induce CD apoptosis, and the dosage-effect relationship of SNP in inducing CD apoptosis was determined. Serum samples from the blank, control, and three dosages of TGXTC-treated rabbits were tested in the CD culture in the presence of SNP. Cell apoptosis was determined by Hoechst 33342 staining, viability of CDs was quantified by MTT, CD apoptosis rate was determined by annexin V-FITC/PI staining, levels of p53 and Bcl-2 mRNA expression in CDs were determined with RT-PCR, and contents of caspase-3 and caspase-9 proteins were determined by colorimetry.</p><p><b>RESULTS</b>CD apoptosis was induced by SNP at all concentrations tested and in a dose-dependent manner. The SNP concentration of 1 mmol/L and treatment duration of 24 h appeared to be optimal and were selected for the study. Serum samples from the positive control rabbits and from the two higher doses of TGXTC-treated rabbits showed reduction of SNP-induced CD apoptosis, decrease in p53 mRNA expression, inhibition of catalytic activities of caspase-3 and caspase-9, and increase in Bcl-2 mRNA expression when compared with the serum from the blank group (P<0.05).</p><p><b>CONCLUSION</b>TGXTC-containing sera antagonized SNP-induced CD apoptosis and the molecular basis for the action was associated with up-regulation of Bcl-2, down-regulation of p53 expression, and inhibition of caspase-3 and caspase-9 catalytic activities.</p>

Effects of time factors on acupoint sticking therapy for preventing and treating bronchial asthma / 中国针灸

<p><b>OBJECTIVE</b>To observe the influence of time factors on acupoint sticking therapy for preventing and treating bronchial asthma.</p><p><b>METHODS</b>Seventy-one cases were randomly divided into a dog days group (n= 30), a Sanjiu days group (n=21) and a daily days group (n=20). They were all treated with ginger moxibustion plus acupoint sticking of Chinese medicine at Dazhui (GV 14) and Feishu (BL 13) etc. This treatment was applied once at the beginning of the first dog days, the middle dog days and the last dog days respectively in the dog days group, and once at the beginning of the first nine days, the middle nine days and the last nine days in coldest days of winter respectively in the Sanjiu days group, and once every other 10 days during 30 days except the dog days or the Sanjiu days in the daily days group. Their therapeutic effects and quality of life and changes of serum level of interleukin 13 (IL-13) were observed.</p><p><b>RESULTS</b>The total effective rate of the dog days group was 83.3% (25/30), the Sanjiu days group and the daily days group were 61.9% (13/21) and 65.0% (15/20) respectively, with no significant differences among three groups (all P>0.05). After treatment, there were no significant differences in quality of life and changes of serum level of IL-13 among three groups, but there were significant differences between before and after treatment (P<0.01, P<0.001).</p><p><b>CONCLUSION</b>Acupoint sticking is an effective therapy for bronchial asthma. It can be practiced in the whole year for the result of this study that medicines and acupoints are the leading factors of this therapy and the time factors have no influence on therapeutic effect.</p>

Effect of Different Carbon Sources on Succinic Acid Production of Actinobacillus succinogenes and Metabolic Flux Analysis / 微生物学通报

In anaerobic bottles fermentation,glucose,fructose,xylose,lactose,maltose,sucrose and sugar alcohols could be used to produce succinic acid with Actinobacillus succinogenes. When sorbitol was utilized as the carbon source in the batch fermentation,more succinate and ethanol were produced compared with those using glucose,while producing less acetate and formate. The metabolic flux analysis results showed that the flux partitioning at PEP node was stable when glucose was replaced by sorbitol,but the flux partitioning at PYR and AcCoA nodes changed a lot because more reducing power(NADH) was generated to meet the more requirement the synthesis of succinate and ethanol.

Breeding Actinobacillus succinogenes with Acid-tolerance by Genome Shuffling / 微生物学通报

A strain Actinobacillus succinogenes CGMCC 1593 was selected as the parent strain.After UV-EMS and UV-DES treatments respectively,seven mutated strains with subtle improvements in acid tol-erance were obtained,and were subjected for recursive protoplast fusion.Through three rounds of genome shuffling,four shuffled strains with both higher yield and acid tolerance were obtained.The shuffled strain namely F3-21 could even survive at pH 5.2.The comparison of the shuffled strains and the parent strain for succinic acid production was also studied here.After 48 h of shake-flask fermentation,the succinic acid concentration of F3-21 was 48% higher than that of the parent strain.When F3-21 was carried out in a 5 liter stirred bioreactor with pH controlled 5.6~6.0,the accumulation of succinic acid in 48 h fermentation attained 38.1 g/L,which was increased by 45% compared with that of the parent strain(26.2 g/L).While pH was controlled at 6.5~7.0,the production of succinic acid in 32 h fermentation attained 40.7 g/L.When F3-21 was carried out in fed-batch fermentation,succinic acid concentration of 67.4 g/L was reached in 72 h fer-mentation.These results indicated that the genome shuffling could improve the acid tolerance and the suc-cinic acid production of A.succinogenes CGMCC 1593.

Inflammatory reaction versus endogenous peroxisome proliferator-activated receptors expression, re-exploring secondary organ complications of spontaneously hypertensive rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The chronic pathological changes in vascular walls of hypertension may exert destructive effects on multiple organ systems. Accumulating evidence indicates that inflammatory reactions are involved in the pathological changes of hypertension. Three peroxisome proliferator-activated receptors (PPARs) have been identified: PPARalpha, PPARbeta/delta, and PPARgamma, all of which have multiple biological effects, especially the inhibition of inflammation. The aim of this study was to evaluate PPAR isoforms expression profile in important organs of spontaneously hypertensive rats (SHR) and to understand the modulation of endogenous PPAR isoforms under inflammatory condition.</p><p><b>METHODS</b>Tissues (kidney, liver, heart, and brain) were dissected from SHR and age-matched control Wistar-Kyoto rats (WKY) to investigate the abundance of PPAR isoforms and PPAR-responsive genes (acyl-CoA oxidase and CD36). The expression of CCAAT/enhancer-binding protein delta (C/EBPdelta), which can trans-activate PPARgamma expression, was also observed. The inflammatory response was analyzed by the expression of inflammatory mediators inducible nitric oxide synthase (iNOS), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin, interleukin-1 beta (IL-1beta), and tumor necrosis factor alpha (TNFalpha), and formation of carbonyl and nitrated proteins.</p><p><b>RESULTS</b>The expressions of 3 PPAR isoforms and PPAR-responsive genes were markedly upregulated in SHR compared with those of WKY. Specifically, the expression of PPARalpha protein in the kidney, liver, heart and brain increased by 130.76%, 91.48%, 306.24%, and 90.70%; PPARbeta/delta upregulated by 109.34%, 161.98%, 137.04%, and 131.66%; PPARgamma increased by 393.76%, 193.17%, 559.29%, and 591.18%. In consistent with the changes in PPARgamma, the expression of C/EBPdelta was also dramatically elevated in SHR. Inflammatory mediators expressions were significantly increased in the most organs of SHR than WKY. As a consequence, increased formation of carbonyl and nitrated proteins were also observed in the most organs of SHR.</p><p><b>CONCLUSIONS</b>These findings suggest an enhanced inflammatory response in the organs of SHR, which might play a key role in pathogenesis of hypertension and secondary organ complications. Changes (increases) in PPARs expression may reflect a compensatory mechanism to the inflammatory status of hypertensive rats.</p>

Environmental factors affecting the succinic acid production by Actinobacillus succinogenes CGMCC 1593 / 生物工程学报

Actinobacillus succinogenes is a promising candidate for the production of bio-based succinic acid. Previously, we isolated a succinic acid-producing strain Actinobacillus succinogenes CGMCC 1593 from bovine rumen. In this paper, the influence of the environmental factors such as gas phase, pH, ORP, on succinic acid production by A. succinogenes CGMCC 1593 was studied. The results showed that CO2 was the optimum gas phase for anaerobic fermentation ofA. succinogenes CGMCC 1593 as well as one of the substrate for the succinic acid synthesis. Using MgCO3 as a pH regulator, the pH was maintained within 7.1-6.2 during the anaerobic fermentation for the cell growth and acid production of A. succinogenes CGMCC 1593. Our results showed that low initial ORP was disadvantageous for the growth of A. succinogenes CGMCC 1593 and an ORP of -270 mV was demonstrated to be beneficial to the succinic acid production. By adding Na2S.9H2O to decrease ORP to -270 mV at the end of exponential growth phase in batch culture of A. succinogenes CGMCC 1593, the succinic acid concentration reached 37 g/L and the yield of succinic acid was 129% at 48 h. This work might provide valuable information for further optimization of succinic acid fermentation by A. succinogenes CGMCC 1593.

Breeding of monofluoroacetate-resistant strains of Actinobacillus succinogenes and the mechanism based on metabolic flux analysis / 生物工程学报

Succinic acid has received a great deal of attention as an important green chemical stock for the manufacture of synthetic resins, biodegradable polymers and chemical intermediates. In this paper, the breeding mechanism of Actinobacillus succinogenes based on metabolic flux analysis was demonstrated to improve the yield of succinic acid by fermentation. After the NTG treatment, mutants from A. succinogenes CGMCC 1593 which were able to grow in medium containing concentrations of about 50-100 mmol/L of sodium monofluoroacetate were obtained. Among them, a mutant SF-9 was selected for producing more succinic acid and less acetic acid. When fermentations were conducted in a 5 L bioreactors, the final succinic acid concentration of SF-9 (34.8 g/L) increased 23.4%, and the mass ratio of succinic acid/acetic acid increased from 3.3 to 9 compared with those of the parent strain. Based on the metabolic flux analysis of A. succinogenes, PEP was found to be a key node which has an important effect on the production of succinic acid, and the flux ratio of by-productions (acetic, formic, lactic acid) was influenced by PYR node. Compared with the parent strain, the flux to succinic acid of mutant (A. succinogenes SF-9) was significantly increased, while the flux to by-productions had an obvious decline. Therefore, PEP and PYR are not rigid nodes in the metabolic regulation of A. succinogenes.

Experimental study on the effects of inactivated and un-inactivated pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction on the proliferation of osteoblast cultured in vitro / 中国骨伤

<p><b>OBJECTIVE</b>To study the effect of inactivated and un-inactivated pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction on the proliferation of osteoblast cells (OB)cultured in vitro.</p><p><b>METHODS</b>OB was isolated from the skull of newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion and identified by image analysis under inverted microscope, V-G collagen staining, ALP staining, calcification nod staining etc. After the OB was identified, in activated and un-inactivated pharmaco-serum of diabetic rats fed with Qianggubao decoction of ferent phase (rats were fed with medicine 3 days or 5 days after last fed with medicine 1 hour or 3 hours) and concentration (5%, 10%, 20%) were added to the OB and incubated. After determined times, the effects of the proliferation of osteoblasts were detected by MTT analysis.</p><p><b>RESULTS</b>There was significant difference between un-inactivated pharmaco-serum and inactivated pharmaco-serum on the proliferation of osteoblasts, and un-inactivated serum had stronger effects to improve the proliferation of osteoblasts (P < 0.01 or P < 0.05).</p><p><b>CONCLUSION</b>Un-inactivated and inactivation pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction can influence the proliferation of, and the un-inactivated pharmaco-serum has stronger effects.</p>

Protection of apoptosis of osteoblast cultured in vitro by Morinda Root Polysaccharide / 中国骨伤

<p><b>OBJECTIVE</b>To explore the protection on apoptosis and the mechanism of promoting the cytoactive of osteoblast by Morinda Root Polysaccharide through the observations of the cultured osteoblast in vitro.</p><p><b>METHODS</b>Prepared blood serum with Morinda Root Polysaccharide and Morinda Root aqueous extract and cultured Osteoblast in vitro with it. The second generation osteoblasts in vitro were separated from the cranium of 24-hours newborn SD rat, which were divided into control group (adding only rat serum during cultivation), induction apoptosis group (adding trans-retinoic acid in control group), Morinda Root aqueous extract group (adding serum prepared by Morinda Root aqueous extract in induction apoptosis group) and Morinda Root Polysaccharide group (adding serum prepared by Morinda Root Polysaccharide in induction apoptosis group). Adopting fluorescence microscope, apoptosis detected by flow cytometry and gene expression of Bcl-2 and Bax detected by RT-PCR, to evaluate the effect of Morinda Root Polysaccharide on the course of osteoblast apoptosis.</p><p><b>RESULTS</b>The apoptotic rate of Morinda Root aqueous extract group and Morinda Root Polysaccharide group were significantly lower than that of induction apoptosis group (P < 0.01). The apoptosis ratio of Morinda Root Polysaccharide group was lower than that of Morinda Root aqueous extract group (P < 0.05). Expression level of Bcl-2 mRNA of apoptosis cell: control group > Morinda Root Polysaccharide group > Morinda Root aqueous extract group > induction apoptosis group (P < 0.01). Expression level of Bax mRNA: induction apoptosis group > Morinda Root aqueous extract group > control group > Morinda Root Polysaccharide group (P < 0.01). Bcl-2/Bax: control group > Morinda Root Polysaccharide group > Morinda Root aqueous extract group > induction apoptosis group (P < 0.01).</p><p><b>CONCLUSION</b>Morinda Root can inhibit the apoptosis of osteoblast induced by trans-retinoic acid in some extent. The above role of Morinda Root Polysaccharide is significant better than that of Morinda Root aqueous extract. It is indicated that Morinda Root Polysaccharide is one of the essential component of inhibiting osteoblast apotosis.</p>

Experimental study on the effects of pharmaco-serum of diabetic rats fed with Chinese herbs qianggubao decoction (Chinese characters: see text) on osteoblast culture in vitro / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the optimum phase and dose of pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) on the differentiation and mineralization of osteoblast (OB). METHODS (OB) was isolated from the skull of 10 newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion. After the OB was identified, different kinds of pharmaco-serum of diabetic rats fed with inactive Qianggubao decoction ([Chinese characters: see text]) of different phase (rats were fed with medicine three days or five days after last fed with medicine one hour or three hours) and concentration (5%, 10%, 20%) were added to the OB and incubated. After 7 days and 18 days of culture,the effects of the differentiation and mineralization of osteoblast were detected.</p><p><b>RESULTS</b>The secretion of ALP and formation of mineralized nodules of osteoblast in the different doses of pharmaco-serum groups were almost the same as that of normal control group, but were superior to that in the model control group. And the group with concentration of 20% pharmaco-serum was the best in the secretion of ALP and formation of mineralized nodules of osteoblast. As to the phases of pharmaco-serum, the best one on the differentiation and mineralization of osteoblast was the serums from diabetic rat-model fed with Qianggubao decoction ([Chinese characters: see text]) three days or five days, after one hour of last fed with medicine.</p><p><b>CONCLUSION</b>The pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) can promote the differentiation and mineralization of osteoblast. Allow for time and the cost of experiment,we presume that pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) three days, after one hour of last fed, with concentration of 20% and not-inactivation is the optimum on the differentiation and mineralization of osteoblast.</p>

Experimental study on effect of Xuefu Zhuyu Decoction on bone marrow hematopoietic stem cells of mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Xuefu Zhuyu Decoction (XFZYD) on the number, phenotype, cell cycle and colony formation of bone marrow hematopoietic stem cells (HSC) in mice.</p><p><b>METHODS</b>Kunming mice were randomly divided into 4 groups: the control group, the low- (3.25 g/kg), middle- (6.5 g/kg) and high-dose (13.0 g/kg) XFZYD groups. After they were medicated by gastrogavage respectively with saline or corresponding dose of XFZYD for 7 days, their bone marrow HSC were separated and counted. The phenotype Sca and cell cycle of HSC were detected by flow cytometer, and the colony formation was determined with semisolid methyl media culture.</p><p><b>RESULTS</b>No obvious difference in the number of mononuclear cell, suspended cell and colony production was found among all the groups (P > 0.05); while the expression of CD34 and Sca-1 increased in the low-dose XFZYD group, but in the middle-dose XFZYD group increase only showed in Sca-1 expression.</p><p><b>CONCLUSION</b>XFZYD plays a role of removing blood stasis and promoting regeneration through improving hematopoietic function by means of increasing the number and enhancing the function of premature HSC.</p>