Objective To establish a HPLC method to determine the content of acteoside in Conghuang Bushen Capsule. Methods The HPLC method was employed with a column of Hypersil ODS2(4.6 mm? 250 mm,5 ? m) and a mobile phase of acetonitrile-methanol-1 % HOAc(13 ∶ 9 ∶ 78),the detection wavelength being at 334 nm,flow rate being 1.0 mL/min and the column temperature being at 40 ℃ . Results The linear range was 0.108 5 ? g ～ 0.650 7 ? g,r=0.999; 1.The average recovery of acteoside and RSD were 98.42 % and 1.89 %,respectively. Conclusion The present method is simple,rapid and reproducible. It can be used for quality control of Conghuang Bushen Capsule.

Cochlear Implantation ; methods ; Cochlear Implants ; Humans ; Minimally Invasive Surgical Procedures

In the past two decades,with the increase of smoking population,more and more people are suffering from small cell lung cancer(SCLC).Besides,it is difficult to find an effective way to cure SCLC,since patience can easily develop drug resistance.On the other hand,with the development of science and technology,people began to study the anti-cancer strategy to increase apoptosis,such as inhibiting the overexpression of survival factors.In these survival factors,BCL-2 family has attracted a lot of attention.BH3-only protein is a member of BCL-2 family and it can directly inhibit the expression of BCL-2 protein,thereby prompting apoptosis.Since the BH3-only protein itself is difficult to become a clinical drug, to find alternatives BH3-only protein-BH3 mimetics is particularly important. Plus, more and more researchers have paid attention on the natural BH3 mimetic since it has less side-effect than artificial BH3 mimetics.To find possible BH3 mimetics,we made a primary screening with this pharma-cophore on a small molecular compounds library via Discovery Studio software. And then MTS assay were introduced to verify the activity of compounds. After that, we use Western Blot and Co-IP meth-ods to test the effect of BH3 mimetics.And finally use CDOCKER to predict the further mechanism on autophagy and apoptosis.In our studies, we found 3 possible BH3 mimetics compounds from 170,000 natural small molecular compounds via pharmacophore-based virtual screening.Furthermore,we dem-onstrated AD23,one of the 3 possible natural BH3 mimetics,induced autophagy and apoptosis simulta-neously in dose-time dependence in SCLC cell line. Finally, we use Molecular Docking to predict the further mechanism on autophagy and apoptosis. We believe our works would provide evidences and clues for the structural optimizing and further study of new drugs in the future.

Objective:To study the effects of gossypol acetic acid(GAA)on the methylation level and the mRNA expression of hM-LH1 in human tongue cancer Tca8113 cells.Methods:Tca8113 cells were treated by GAA at various doses for 24 h,48 h and 72 h respectively.MTT assay was used to detect the cell proliferation.Nested methylation specific PCR(nMSP)was used to detect methyl-ation level of hMLH1 .Real-time fluorescence quantitative PCR(RFQ-PCR)was applied to investigate the mRNA expression of hM-LH1 gene.Results:GAA inhibited the proliferation of Tca8113 cells dose-and-time dependently,decreased the DNA methylation level of hMLH1(P<0.05)and increased hMLH1 mRNA expression in Tca8113 cells(P<0.05).Conclusion:GAA can suppress proliferation of Tca8113 cells by demethylation of hMLH1 gene and increase of hMLH1 mRNA expression.

Objective To setup a measurement of human bone marrow micromegakaryocyte which based on CD41a and PI double‐labeled flow cytometric analysis ,and study the significance in the diagnosis of MDS .Methods In 42 cases of MDS patients , their bone marrow megakaryocytes were obtained by Percoll density gradient separation medium .The megakaryocyte glycoproteinⅡb/Ⅲa(CD41a)were marked with fluorescein isothiocyanate through its corresponding monoclonal antibody ,and their DNA were marked with PI .Then the megakaryocyte ploidy was analyzed by flow cytometry(FCM ) .Results The method for micromegakaryo‐cyte identification and analysis was established .In 42 patients with MDS ,the detection rate of micromegakaryocyte was 90 .5 per‐cent by FCM analysis ,but only 54 .8 percent by Wright‐Giemsa staining test and 64 .3 percent by immunohistochemistry ,the differ‐ence among them was statistically significant(χ2 = 13 .640 ,P= 0 .001) .The 42 patients with MDS were divided into two groups (low‐risk group and high‐risk group) .The detection rates of micromegakaryocyte were 81 .8 percent in low‐risk group and 100 per‐cent in high‐risk group separately by FCM analysis ,the difference was statistically significant(χ2 =4 .019 ,P=0 .045) .Conclusion The detection rate of micromegakaryocyte by FCM with CD41a and PI double marker is higher than that by cytochemical staining . The detection rate of micromegakaryocyte in the high‐risk group is higher than that of the low‐risk group ,which shows that the de‐tection of micromegakaryocyte is of great significance for MDS prognosis assessment .

Objective The purpose of this study was to investigate the effects of gossypol acetic acid (GAA) on protein and mRNA expressions of hMLH1 gene in human tongue carcinoma cell line Tca8113 in vitro in order to discuss the mechanism of tumor suppression of GAA. Methods (1) Western-blot was used to study the effects of GAA on protein expressions of hMLH1 gene in Tca8113 cell line treated by different concentrations of GAA for 48 h. (2) Real-time fluorescence quantitative PCR (RFQ-PCR) was used to investigate the effects on the mRNA expressions of hMLH1 gene in Tca8113 cell line treated by GAA for 48 h. Results (1) Compared with the control group, the results of Western-blot showed that the protein expression of hMLH1 gene was increased after treatment by GAA for 48 h ( <0.05) . (2) The results of RFQ-PCR indicated that the mRNA expression of hMLH1 gene was increased after GAA treatment for 48 h ( <0.05) . Conclusion GAA could up-regulate protein and mRNA expression of hMLH1 in Tca8113 cell line, which indicated that it may be one of the mechanisms of tumor suppression effect of GAA.

Objective:To observe the effect of Rhei Radix et Rhizoma and Persicae Semen on inflammatory cytokines and intestinal mucosal barrier in rats with postoperative adhesive intestinal obstruction, and to explore its mechanism. Methods:Sixty SD rats were randomly divided into normal group, sham operation group, model group, low, medium and high dose of Rhei Radix et Rhizoma and Persicae Semen group. Except for the normal group and the sham operation group, and the other animals groups were established the model of postoperative adhesive intestinal obstruction. Rhei Radix et Rhizoma and Persicae Semen low, medium and high dose groups were perfused with Rhei Radix et Rhizoma and Persicae Semen decoction with the concentration of 0.2, 0.6 and 1.8 g/ml, the normal group, sham operation group and model group were gavaged with equal volume of sterile saline from the first day after the operation, once a day. After corresponding treatment, the adhesion score was observed on the 7th day after the operation, the contents of interleukin-1 β (IL-1β), D-lactic acid (D-LA), diamine oxidase (DAO) and vascular endotoxin (ET) in serum were detected by ELISA method, and the expression of SIgA, CD4 +T and CD8 +T cells in intestinal mucosa were assessed by immunohistochemical method. Results:Compared to the model group, the adhesion score in the low, medium and high dose of Rhei Radix et Rhizoma and Persicae Semen group significantly decreased ( P<0.05), the level of serum IL-1β [(8.66 ± 1.07) ng/L, (8.15 ± 1.23) ng/L, (7.99 ± 1.11) ng/L vs. (14.08 ± 2.54) ng/L] significantly decreased ( P<0.01), the expression of SIgA (1.38 ± 0.15, 2.87 ± 1.17, 2.79 ± 0.80 vs. 0.65 ± 0.12) in intestinal mucosa in the low, medium and high dose of Rhei Radix et Rhizoma and Persicae Semen group significantly increased ( P<0.01). The levels of serum D-LA [ (8.57 ± 1.73) mg/L, (7.13 ± 1.75) mg/L vs. (14.58 ± 2.81) mg/L], ET [ (77.39 ± 6.83) mg/ml, (50.49 ± 7.80) mg/ml vs. (138.22 ± 7.79) mg/ml] significantly decreased ( P<0.01), the expression of CD4 +T (2.61 ± 0.83, 2.91 ± 1.62 vs. 1.15 ± 0.98) and CD8 +T (2.88 ± 0.69, 3.01 ± 1.86 vs. 1.26 ± 0.74) cells in intestinal mucosa in the medium and high dose of Rhei Radix et Rhizoma and Persicae Semen group significantly increased ( P<0.01). Conclusion:Rhei Radix et Rhizoma and Persicae Semen decoction can improve intestinal mucosal permeability, protect intestinal mucosal immune barrier and reduce inflammatory reaction in the treatment of adhesive intestinal obstruction.

Objective:By applying moxibustion to the eight confluent points in different periods of time,to observe the changes in thermal pain threshold latency of acupoints based on Fei Teng Ba Fa.Methods:A total of 468 healthy college student volunteers received moxibustion at the eight confluent points in three different periods of time,i.e.Chen (7:00-9:00),Wu (11:00-13:00) and Xu (19:00-21:00).The thermal pain threshold latency was adopted to measure the changes in pain threshold of the eight confluent points under different conditions (different periods of time,different genders,different acupoints and different states of the acupoints) based on Fei Teng Ba Fa.Results:Finally,thirty subjects dropped out and 438 subjects were included.The comparison of thermal pain threshold latencies of the eight confluent points in the same opening or closing state based on Fei Teng Ba Fa:latencies of the closing points and adjunct points were significantly different in different periods of time (P＜0.05);the latencies of the males were significantly longer than those of the females (P＜0.05);there was no significant difference in the latency between the left and right sides (P＞0.05);in the female group,there was a significant difference in the latency between the lower-limb points and the upper-limb points (P＜0.05).The comparison of thermal point threshold latencies of the eight confluent points in different opening or closing state:in the period of Wu (11:00-13:00),the latencies of the opening points were significantly longer than those of the closing points and adjunct points (P＜0.05);for men,their opening and closing points had significantly longer thermal pain threshold latencies than their adjunct points (P＜0.05);despite the gender,the latencies of the upper limb opening and closing points were significantly longer than the latency of the adjunct points (P＜0.05);in the female group,the latencies of the lower-limb opening points were significantly shorter than those of the lower-limb closing and adjunct points (P＜0.05).Conclusion:Based on Fei Teng Ba Fa,the pain thresholds of the eight confluent points vary in different periods of time,gender,acupoint location and opening/closing state,which can be taken as the evidence of making time-based acupuncture-moxibustion prescriptions.

AIM: Research on the change of the saponins constituents of Radix Bupleuri after being processed has been carried out. METHODS: Saikosaponin a、c、d and b2 are used as the marker ingredients; the change of saponins constituents,both after procession and saponated action,have been determined by HPLC. RESULTS: After being processed,the content of saikosaponin b2 has a significant increase,saikosaponin a,saikosaponin c,saikosaponin d and saikosaponin a + c + d all have slight decrease. After the saponated action,the content of saikosaponin a + c + d has a little change,and saikosaponin b2 has increased significantly. CONCLUSION: The change rules of saponin compounds in processed Radix Bupleuri have been revealed.

Objective To study if 5-Aza-2’-deoxycytidine along or together with 4-phenylbutyric acid could affect miR-196b expression levels in chronic myeloid leukemia cells .Methods K562 cells were treated with DNA methylation inhibitor 5-Aza-2’-deoxycytidine, histone deacetylase inhibitors 4-phenylbutyric acid separately and the combined treatment with both of them, then expression levels of miR-196b were detected using Real-time PCR.Results The half inhibition concentration of 4-phenylbutyric acid was 1.58mmol/L.Comparing with the expression level of miR-196b in normal human bone marrow cells, the expression levels of miR-196b were significantly lower in Aza group , PBA group and negative control cells and nearly consistent among three groups , and as high as normal cells in combined treatment group . Conclusion The expression level of miR-196b in K562 cells could not return to normal treated with 5-Aza-2 ’-deoxycytidine or 4-phenylbutyric acid separately , while could restore normal when treated with both agents , indicating that miR-196b expression level in K562 cells is related with both DNA methylation and histone acetylation .