Cloning and quantitative expression analysis of GMPP gene from Dendrobium huoshanense / 中国中药杂志

In order to understand the function of GDP-mannose pyrophosphorylase(GMPP) function and its regulation in polysaccharide biosynthesis mechanism in Dendrobium. D. huoshanense was used to clone GMPP gene. GMPP gene expression in D. huoshanense,D. officinale and D. moniliforme was also determined by qPCR. The results showed that the length of D. huoshanense GMPP gene c DNA sequence is 1 867 bp,containing 1 245 bp open reading frame(ORF),encoding 415 amino acids. Phylogenetic tree analysis showed that D. huoshanense,D. officinale and D. moniliforme are closely related with GMPP taken into consideration. Bioinformatics analysis demonstrated that GMPP sequence similarity among the three species reached as high as 99%. qPCR results indicated that GMPP genes was highly expressed in stem of D. huoshanense compared with its leaf,flower and root. According to GMPP gene expression profile in D. huoshanense,D. officinale and D. moniliforme grown in Huoshan area,it was clear that GMPP in D. huoshanense showed the highest expression level. Furthermore,our findings of GMPP gene expression profile will facilitate future researches into its polysaccharide biosynthetic mechanism.

Clinical Study on Electronic Moxibustion in Treating Diabetic Peripheral Neuropathy / 上海针灸杂志

Objective To observe the clinical efficacy of electronic moxibustion in treating diabetic peripheral neuropathy due to liver-kidney deficiency. Method Ninety patients with diabetic peripheral neuropathy due to liver-kidney deficiency were randomized into a treatment group of 44 cases and a control group of 46 cases. The treatment group was intervened by acupoint therapy by warm electronic moxibustion, while the control group was intervened by foot bath with Chinese medication. For both groups, 10 treatment sessions were taken as a course of treatment, for 3 courses in total. Before and after the treatment, the whole blood viscosity, blood lipids, fasting blood glucose (FBG), glycosylated hemoglobin (GHb) and Michigan Neuropathy Screening Instrument (MNSI) were evaluated in the two groups. Result The whole blood viscosity indexes, blood lipids levels, FBG level and GHb level didn't show significant differences after the treatment in the two groups (P>0.05). The intra-group and inter-group comparisons of MNSI score showed significant differences after the treatment (P<0.01); there was a between-group difference in the change of MNSI score after the treatment (P<0.01). Conclusion Electronic moxibustion is an effective approach in treating diabetic peripheral neuropathy.

Observations on the Therapeutic Effect of Electronic Moxibustion on Diabetic Peripheral Neuropathy / 上海针灸杂志

Objective To investigate the clinical efficacy of electronic moxibustion in treating diabetic peripheral neuropathy (liver-kidney deficiency type).Methods Ninety patients with diabetic peripheral neuropathy (liver-kidney deficiency type) were randomized to a treatment group (44 cases) and a control group (46 cases).The treatment group received electronic moxibustion and the control group,Chinese herbal fumigation of feet.Both groups were treated 10 times as a course for a total of three courses.The TCM symptom total score and the Michigan Neuropathy Screening Instrument (MNSI) score were recorded in the two groups before and after treatment.The clinical therapeutic effects were compared between the two groups.Results There was a statistically significant pre-/post-treatment difference in the TCM symptom total score in both groups after one,two and three courses of treatment (P＜0.01).The TCM symptom total score improvement rate was (60.9±16.7)％ in the treatment group and (56.6±19.2)％ in the control group;there was a statistically significant difference between the two groups (P＜0.01).The total efficacy rate was 97.7％ in the treatment group and 95.7％ in the control group;there was no statistically significant difference between the two groups (P＞0.05).The pre-/post-treatment MNSI score difference value was (2.98±3.66) in the treatment group and (0.78±2.92) in the control group;there was a statistically significant difference between the two groups (P＜0.01).Conclusions Electronic moxibustion is an effective way to treat diabetic peripheral neuropathy.Its therapeutic effect is slightly better than that of Chinese herbal fumigation.

MicroRNA-210 Plays a Critical Role in the Angiogenic Effect of Isoprenaline on Human Umbilical Vein Endothelial Cells via Regulation of Noncoding RNAs / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>β-adrenoceptors play a crucial regulatory role in blood vessel endothelial cells. Isoprenaline (ISO, a β-adrenergic agonist) has been reported to promote angiogenesis through upregulation of vascular endothelial growth factor (VEGF) expression; however, the underlying mechanism remains to be investigated. It is widely accepted that certain noncoding RNAs, including microRNAs (miRNAs) and long noncoding RNAs (lncRNAs), can regulate endothelial cell behavior, including their involvement in angiogenesis. Therefore, we aimed to investigate whether noncoding RNAs participate in ISO-mediated angiogenesis using human umbilical vein endothelial cells (HUVECs).</p><p><b>METHODS</b>We evaluated VEGF-A messenger RNA (mRNA) and protein levels in ISO-treated HUVECs by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. To establish whether noncoding RNAs are associated with ISO-mediated angiogenesis, we measured expression of the miRNAs miR-210, miR-21, and miR-1, as well as that of the lncRNAs growth arrest-specific transcript 5 (GAS5), maternally expressed 3 (MEG3), and metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in HUVECs exposed to ISO. Furthermore, to ascertain its importance in ISO-mediated angiogenesis, we constructed the HUVECs with overexpressing miR-210 and detected the subsequent expression of VEGF-A and noncoding RNAs. All statistical analyses were performed using SPSS 16.0 software. Intergroup comparisons were carried out by one-way analysis of variance.</p><p><b>RESULTS</b>VEGF-A mRNA levels were elevated in the ISO group (1.57 ± 0.09) compared to those in the control group (P < 0.01). Moreover, concentrations of VEGF-A in culture supernatants significantly differed between the control (113.00 ± 19.21 pg/ml) and ISO groups (287.00 ± 20.27 pg/ml; P< 0.01). Expression of miR-1, miR-21, and miR-210 was higher (3.89 ± 0.44, 2.87 ± 087, and 3.33 ± 1.31, respectively) in ISO-treated cells than that in controls (P < 0.01), whereas that of GAS5 and MEG3 (0.22 ± 0.10 and 0.58 ± 0.16, respectively) was lower as a result of ISO administration (P < 0.05). There was no significant difference in the expression of MALAT1 between the groups. Interestingly, miR-210 overexpression heightened the levels of VEGF-A and miR-21 (5.87 ± 1.24 and 2.74 ± 1.15, respectively; P< 0.01) and reduced those of GAS5 and MEG3 (0.19 ± 0.01 and 0.09 ± 0.05, respectively; P< 0.01).</p><p><b>CONCLUSIONS</b>ISO-mediated angiogenesis was associated with altered expression of miR-210, miR-21, and the lncRNAs GAS5 and MEG3. The effects of miR-210 on the expression of VEGF-A and noncoding RNAs were similar to those of ISO, indicating that it might play an important role in ISO-mediated angiogenesis.</p>

Negative Association of Circulating MicroRNA-126 with High-sensitive C-reactive Protein and Vascular Cell Adhesion Molecule-1 in Patients with Coronary Artery Disease Following Percutaneous Coronary Intervention / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Percutaneous coronary intervention (PCI) causes endothelial damage, resulting in an inflammatory response with elevation of markers such as high-sensitive C-reactive protein (hs-CRP) and vascular cell adhesion molecule-1 (VCAM-1), which are associated with restenosis after PCI. Evidence suggests that microRNA-126 (miR-126) plays an important role in vascular inflammation, but its correlation with PCI-mediated inflammation has not been investigated. In this study, we investigated the effect of PCI on circulating miR-126 and inflammation markers such as hs-CRP and VCAM-1.</p><p><b>METHODS</b>We enrolled 130 patients with coronary artery disease (CAD) in the Second Hospital of Jilin University from October 2015 to December 2015. Among them, 82 patients with CAD, defined as at least one major epicardial vessel with >70% stenosis who planned to undergo PCI, were divided into acute coronary syndrome (ACS) group (46 patients) and stable angina (SA) group (36 patients). Forty-eight patients confirmed by coronary angiography without PCI were used as controls. The plasmas of all patients were collected prior to PCI and at 30 min, 24 h, and 72 h after PCI. The plasma VCAM-1 and hs-CRP were detected by enzyme-linked immunosorbent assay, and the miR-126 was evaluated by quantitative reverse transcription-polymerase chain reaction.</p><p><b>RESULTS</b>Plasma concentrations of hs-CRP and VCAM-1 in patients with either ACS (n = 46) or SA (n = 36) were significantly higher than in controls (n = 48) (P < 0.01) prior to PCI, and increased further at 24 h and 72 h after PCI, compared with prior PCI. Moreover, VCAM-1 was positively correlated with balloon time and pressure. In contrast, the plasma concentration of miR-126 was significantly lower in patients with CAD than in controls, and further decreased with time post-PCI. A negative correlation was observed between miR-126 and hs-CRP and VCAM-1 at 72 h after PCI.</p><p><b>CONCLUSION</b>There was a negative correlation of miR-126 with the PCI-induced markers of inflammation such as hs-CRP and VCAM-1.</p>

Heterogenous Gene Expression of Methyl Parathion Hydrolase and Analysis of the Enzyme Activity / 微生物学通报

Methyl parathion hydrolase (MPH, E.C.3.1.8.1) coding gene mph from Pseudomonas sp. WBC-3, isolated and identified by our lab, was successfully expressed in E. coli AD494 (DE3)/ pET32a(+) system as soluble fusion form at high level. The recombinant MPH showed nearly 4～5 fold higher specific activity to parathion than the enzyme from Pseudomonas sp. WBC-3. In addition, the thermal stability of the recombinant enzyme was improved comparing with the wild type enzyme.