Clinical observation on the neurotransmitters regulation in patients of insomnia differentiated as yang deficiency pattern treated with warm acupuncture and auricular point sticking therapy / 中国针灸

<p><b>OBJECTIVE</b>To evaluate the effect mechanism of warm acupuncture combined with auricular point sticking therapy and its efficacy on insomnia by monitoring the level of brain neurotransmitters in the insomnia patients.</p><p><b>METHODS</b>One hundred and thirty patients with insomnia were randomized into an observation group and a control group, 65 cases in each one. In the observation group, based on the treating principle of warming yang and benefiting qi, acupuncture was applied to Xinshu (BL 15), Pishu (BL 20), Shenshu (BL 23), Yaoyangguan (GV 3), Baihui (GV 20), Neiguan (PC 6) and Shenmen (HT 7). Warm acupuncture was supplemented at Xinshu (BL 15), Pishu (BL 20), Shenshu (BL 23) and Yaoyangguan (GV 3). The treatment was given once every day. In the control group, estazolam tablets, 0. 5 to 1 mg were prescribed for oral administration, 30 min before sleep at night. The treatment of 14 days was taken as one session and 2 sessions were required in the two groups. The encephal of luctuograph technology was used to observe the sleep quality and brain neurotransmitters before and after treatment in the two groups and the efficacy was compared between the two groups.</p><p><b>RESULTS</b>The total effective rate was 83.1% (54/65) in the observation group and 87.7% (57/65) in the control group. The efficacy was similar between the two groups (P > 0.05). In the observation group, after treatment, 5-HT and GABA/Glu were all increased compared with those before treatment (P < 0.05, P < 0.01) and norepinephrine (NE) was reduced compared with that before treatment (P < 0.05). The level of each index did not change significantly before and after treatment in the control group (all P > 0.05). The regulations of 5-HT, GABA/Glu in the observation group were superior to those in the control group (all P < 0.05).</p><p><b>CONCLUSION</b>The combined therapy of warm acupuncture and auricular point sticking method for warming yang and benefiting qi effectively improves brain neurotransmitters and essentially improves sleep quality of the patients with insomnia differentiated as yang deficiency pattern.</p>

Clinical observation on strong stimulation of acupoints around eyes for treatment of adolescent myopia / 中国针灸

<p><b>OBJECTIVE</b>To observe the therapeutic effect and influential factors of strong stimulation of acupoints around eyes by reducing method of twisting the needle for treatment of adolescent myopia, and to compare the different curative effects of both weak and strong stimulus intensities and wearing glasses in daytime.</p><p><b>METHODS</b>According to the principle of knowing and permission, the patients were divided into a strong stimulation group (60 cases, 120 eyes), a weak stimulation group (60 cases, 120 eyes) and a wearing glasses group (60 cases, 120 eyes). Both the strong stimulation group and the weak stimulation group were treated by reducing method of twisting the needle, with Cuanzhu (BL 2), Yuyao (EX-HN 4), Sizhukong (TE 23), Sibai (ST 2) and Taiyang (EX-HN 5) selected. Each acupoint was twisted 360 and 90 times per min for the strong stimulation group, and 90 degrees and 60 times per min for the weak stimulation group; the wearing glasses group were not treated with acupuncture, but with wearing glasses in daytime.</p><p><b>RESULTS</b>After treatment with the reducing method of twisting the needle, the vision improved (P < 0.01), with more obviously increased in the strong stimulation group than the weak stimulation group (P < 0.01), and there was no significant change of vision in the wearing glasses group. The age and the myopic degree of the patient before treatment were closely related with the change of vision after treatment (P < 0.01).</p><p><b>CONCLUSION</b>The reducing method of twisting the needle has a definite clinical therapeutic effect on adolescent myopia; strong stimulation has a better effect than the weak one; the age and the myopic degree of the patient possibly influence the therapeutic effect.</p>

Comparison between protein detection methods for two-dimensional gel electrophoresis / 中国实验血液学杂志

The aim of this study was to compare and analyze protein staining in order to select the optimal staining method for proteomic research. Proteins from acute promyelocytic leukemia cell line NB4 and protein molecular weight marker were separated respectively by 2-D or 1-D electrophoresis and detected respectively by the typical Coomassie brilliant blue, the colloidal Coomassie brilliant blue, the modified Coomassie brilliant blue and the silver staining protocols. The protein detection sensitivity, compatibility with mass spectrometry (MS) and facility of the four staining protocols were compared. The results indicated that the silver staining exhibited the highest sensitivity and MS showed the lowest compatibility 10% of protein identification rate. The detection sensitivity of the modified Coomassie brilliant blue staining was superior to that of other two Coomassie brilliant blue stainings, close to but lower than the silver staining, however the compatibility with MS was better (protein identification rate about 55%). It is concluded that the protein detection sensitivity of the modified Coomassie brilliant blue staining is high, and its compatibility with MS is better, this modified Coomassie brilliant blue staining is an optimal staining method for proteomic research.

Prokaryotic expression of extracellular ligand binding domains of chick tie-2 and its anti-angiogenesis effect / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the prokaryotic expression of extracellular ligand binding domains of chick tie-2, the purification, refolding conditions of the recombinant protein, and its anti-angiogeneic effect.</p><p><b>METHODS</b>A DNA fragment encoding extracellular ligand binding domains of chick tie-2 was obtained by PCR amplification using a previous constructed plasmid as a template. The amplified fragment was then inserted into prokaryotic expression vector pQE30, and was expressed in E.Coli XL-1 blue by adding isopropyl-beta-D-thiogalactoside(IPTG). The recombinant protein in inclusion bodies was purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography under denatured conditions. Then the refolding of the purified protein was performed with gradient dialysis. The target protein was injected s.c. into mouse, and the antibody was detected by ELISA and Western blot analysis. The antibody was purified from the antiserum and then incubated with human umbilical endothelial vein cell (HUEVC) to find its anti-angiogenesis in vitro by using propidium iodide(PI) dying through FACS. Alginate encapsulated tumor cell assays were performed and micro-vessel density was determined by counting per high power field in the sections stained with an antibody reactive to CD31 to test its inhibition of angiogenesis.</p><p><b>RESULTS</b>The recombinant protein was highly expressed in E.Coli XL-1 blue, and the antibody produced in mouse could specifically recognize the recombinant protein. The purified antibody could induce apoptosis of HUEVC in vitro. The anti-angiogenic effect of the antibody could also be found in alginate-encapsulate tumor cell assay and by counting micro-vessel density.</p><p><b>CONCLUSION</b>The protein of extracellular ligand binding domains of chick tie-2 can be expressed at high level in the prokaryotic expression system, and the expressed protein can induce immune response in mouse. Furthermore, the antibody can induce the anti-angiogenic effect.</p>

Antisense DNMT1 gene fragment in the sensitivity change of SMMC-7721 cells to tumor necrosis factor related apoptosis inducing ligand and its mechanism / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the sensitivity change of SMMC-7721 cells transfected with antisense DNMT1 gene fragment to tumor necrosis factor related apoptosis inducing ligand (TRAIL) and its mechanism.</p><p><b>METHODS</b>Cell survival rate was measured by trypan blue, apoptosis rate by TUNEL method and the expression of bcl-2, bax and bad by flow cytometry.</p><p><b>RESULTS</b>Cell survival rate of SMMC-7721 cells transfected with antisense DNMT1 gene fragment was markedly lower than that transfected with sense DNMT1 gene fragment or empty vector (P < 0.05 and 0.01), but the apoptosis rate was on the contrary (P < 0.05 or 0.01). The expression of bax and bad (especially the former), but not bcl-2 of SMMC-7721 cells transfected with antisense DNMT1 gene fragment was markedly higher than those of SMMC-7721 cells transfected with sense DNMT1 gene fragment or empty vector.</p><p><b>CONCLUSION</b>The sensitivity of SMMC-7721 cells to TRAIL can be enhanced by the transfection of antisense DNMT1 gene fragment, which may be related to the increase of bax and bad expression.</p>