Determination of deuterium incorporation of new drug donafenib tosilate by two methods of LC-MS with natural isotope abundance correction and 1H NMR relative quantitative method / 药学学报

An LC-MS method with natural isotope abundance correction and a 1H NMR relative quantitative method were established to determine the deuterium incorporation of donafenib tosilate, a new deuterated drug molecule. First, the peak areas of isotopic impurities (non-deuterated and incompletely deuterated impurities) and deuterated drug were recorded through the single ion monitoring (SIM) mode of the established LC-MS method and then corrected in terms of the natural isotope abundance offered by ChemDraw soft, removing the nature isotope interference from 13C, 37Cl, etc. The corrected areas were subsequently used to calculate mol% of isotopologues (D0, D1, D2, D3) and Atom% D, namely, deuterium incorporation. In addition, a 1H qNMR experiment was conducted with the aromatic proton at δ 8.63 and the residual proton of isotopic impurities at δ 2.79 as quantitative peaks. The mixture of DMSO-d6 and D2O (10∶1) was employed as the solvent to change the spin-coupling between the residual proton and active hydrogen so that the residual proton could be measured as the single peak, and the sensitivity was greatly improved. The acquisition parameters were also optimized, and Atom% 1H and the deuterium incorporation were then calculated. The two methods were applied to samples of three commercial batches, and the testing results were almost consistent. Both methods proved accurate, sensitive, fast and independent of standard substances and accurate weighing, which could be applied to the determination of the deuterium incorporation of donafenib tosilate and provide a reference for other deuterated drugs.

Material Attribute Research in Quality Control of Traditional Chinese Medicine Pills： A Review / 中国实验方剂学杂志

As intelligent production of traditional Chinese medicine （TCM） has been inevitable， informatization and automation of the production process have become the precondition for realizing intelligent manufacturing of TCM， of which the accumulation of critical material attribute and the critical quality attribute are the basis. The study of material properties is of great significance to achieve the quality control of the final product in the process， but there is a lack of systematic induction and summary of the research on the attribute of TCM pills. Therefore， the authors analyzed and summarized the attributes of raw materials， excipients and intermediates in the pill unit process according to the classification of powder properties， rheological properties and texture properties. What’s more， the impact of material attributes on the quality of the final product was summarized. Besides， this review summarized the attribute characterization techniques involved in the pill process and provided some suggestions for the characterization of product quality attributes. Finally， based on the concept of quality by design （QbD）， the authors proposed that the study of material attribute should be combined with process analytical technology （PAT）， and the focus of drug quality control should be moved forward to guide equipment upgrading， so as to realize intelligent continuous manufacturing of TCM pills.

Germacranolide sesquiterpenes from Carpesium cernuum and their anti-leukemia activity / 中国天然药物

In this study, three new germacranolide sesquiterpenes (1-3), together with six related known analogues (4-9) were isolated from the whole plant of Carpesium cernuum. Their structures were established by a combination of extensive NMR spectroscopic analysis, HR-ESIMS data, and ECD calculations. The anti-leukemia activities of all compounds towards three cell lines (HEL, KG-1a, and K562) were evaluated in vitro. Compounds 1-3 exhibited moderate cytotoxicity with IC

Inhibitory Effect and Mechanism of Jingulian Capsule on Human Breast Cancer MDA-MB-231 Cells / 中国实验方剂学杂志

Objective:To observe effect of Jingulian capsule on the proliferation of human breast cancer MDA-MB-231 cells and investigate its action mechanism against triple negative breast cancer （TNBC）. Method:The ingredients of Jingulian capsule were identified by ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）. The inhibitory effect of Jingulian capsule at different doses （0.125，0.25，0.5，1，and 2 g·L^-1） against the proliferation of MDA-MB-231 cells were detected by methyl thiazolyl tetrazolium （MTT） assay. After treatment for 24 h， the morphological changes in nuclear apoptosis of MDA-MB-231 cells were detected by Hoechst 33258 staining. The effect of different concentrations of Jingulian capsule on the apoptosis and cycle of MDA-MB-231 cells after different treatment time were determined by flow cytometry. The protein expression levels of Poly-ADP-ribose polymeras （PARP）， proto-oncogene c-Myc， cyclin B₁， and phosphorylated extracellular signal-regulated kinase （p-ERK） in each group were assayed by Western blot. Result:A total of 113 compounds in Jingulian capsule were identified by UPLC-MS/MS. As revealed by MTT assay，compared with blank group，Jingulian capsule （0.125，0.25，0.5，1，2 g·L^-1） significantly inhibited viability of MDA-MB-231 cells （<italic>P</italic><0.01）， with the half maximal inhibitory concentration （ IC₅₀） of（0.13±0.02）g·L^-1. According to flow cytometry，compared with the blank group，Jingulian capsule at 1 g·L^-1 significantly induced the apoptosis of MDA-MB-231 cells （<italic>P</italic><0.05）and Jingulian capsule at 0.5， 1 g·L^-1 obviously increased the number of MDA-MB-231 cells in S phase （<italic>P</italic><0.05，<italic>P</italic><0.01）. The results of Western blotting demonstrated that the protein expression levels of PARP，c-Myc，and cyclin B₁ in 0.5， 1 g·L^-1Jingulian capsule groups were remarkably down-regulated as compared with those in the blank group（<italic>P</italic><0.01）， and the protein expression level of p-ERK in 1 g·L^-1Jingulian capsule group was also down-regulated （<italic>P</italic><0.01）. Conclusion:Jingulian capsule is able to inhibit the proliferation of MDA-MB-231 cells，induce S phase cell cycle arrest， and promote their apoptosis， which may be related to the inactivation of the MAPK signaling pathway.

Protective Effect of Syringaresinol on Excitatory Damage Induced by Sodium Glutamate in SH-SY5Y Cells / 中国实验方剂学杂志

Objective: To establish a model for the injury of human neuroblastoma cell (SH-SY5Y) induced by sodium glutamate, and to observe the protective effect of syringaresinol on cell damage from Viscum liquidambaricolum hayataon, and to explore its mechanism. Method: Construction of SH-SY5Y cell injury model using sodium glutamate.The experiment was divided into normal cell group, injury model group (sodium glutamate 50 mmol·L-1, sodium glutamate 50 mmol·L-1 + DMSO),syringaresinol experimental group (6.25, 12.5, 25 μmol·L-1), by cell counting, cell morphology observation, Annexin V-FITC/PI apoptosis detection, ROS reactive oxygen species detection, mitochondrial membrane potential, and Western blot, evaluation of syringaresinol on glutamate-induced neuronal excitability injury neuroprotective activity. Result: Compared with normal group, the cell survival rate of the model group was significantly decreased (PPPPP-1) showed a concentration-dependent increase in cells. Survival rate (PPPPPConclusion: Syringaresinol has significant protective activity against excitatory damage induced by sodium glutamate in SH-SY5Y neurons, the mechanism may be through anti-oxidative stress, repairing mitochondrial function and DNA damage to significantly reduce sodium glutamate-induced neuronal apoptosis.

Effect of N-cadherin on Proliferation，Migration and Invasion of Retinal Pigment Epithelial Cells / 中山大学学报(医学科学版)

@#【Objective】To clarify the role of neuro-cadherin（N-cadherin）in epithelial-mesenchymal transition of diabetic retinopathy，and to investigate the effect of N-cadherin on proliferation ，migration and invasion of retinal pigment epithelial cells.【Methods】Cells were turned into over-expressed or silenced N-cadherin by using Ad-N-cadherin （Ad-N-cad）and Ad-si N-cadherin（si N-cad）. Glucose（25 mmol/L）was used to simulate high glucose（HG）condi⁃ tions. Cell Counting Kit-8（CCK-8）kit was used to detect cell proliferation. Transwell chamber was used to detect the vertical migration and invasion of cells.【Results】Transwell assay showed N-cadherin over-expression increased the num⁃ ber of cells migrated to the transwell subventricular chamber. The difference was statistically significant（P < 0.05）. The number of ARPE19 cells that migrated to or invaded the transwell subventricular chamber increased after high glucose treatment. N-cadherin knockdown suppressed high glucose-induced migration and invasion（P < 0.05）. CCK8 results showed N-cadherin knockdown could inhibit cell proliferation induced by high glucose（P < 0.05）.【Conclusion】N-cad⁃ herin may promote cell migration，and down-regulation of N-cadherin can inhibit cell proliferation，migration and inva⁃ sion induced by high glucose.

Advances in pharmacological research of sanshool / 中国药理学通报

The pericarp of Zanthoxylum bungeanum Maxim. (Rutaceae) is a traditional Chinese herbal medicine with many therapeutic effects such as antipruritic, analgesic and insecticidal effects. In recent years, various studies have revealed sanshool as the main bioactive ingredient of Zanthoxylum bungeanum Maxim. It can bind to a variety of ion channels and receptors, exerting wide-ranging biological and pharmacological activities including analgesic effects, intestinal protection, hypoglycemic effects and other effects. The relevant pharmacokinetics, drug targets and pharmacological effects were reviewed in this paper to provide a reference for the systematic research of sanshool and the development of new drugs.

Efficacy on borderline hypertension treated with acupuncture combined with anti-hypertensive plaster intervention / 中国针灸

<p><b>OBJECTIVE</b>To compare the efficacy difference in treatment of borderline hypertension between acupuncture combined with anti-hypertensive plaster and simple acupuncture.</p><p><b>METHODS</b>Sixty patients of borderline hypertension were randomized into an observation group and a control group, 30 cases in each one. In the observation group, acupuncture was applied to Fengchi (GB 20), Taichong (LR 3) and the others, once a day, 5 treatments made one session. There were 2 days at interval among the sessions. Totally, 4 sessions were required. Additionally, in combination, the anti-hypertensive plaster was prepared with the fine powder of Chuanxiong (ligusticum wallichii) and Wuzhuyu (fructus evodiae) at 1.1 and mixed with vinegar. The plaster was applied to the umbilicus, once every two days, totally 15 times were required. In the control group, acupuncture was given simply. The selected acupoints and operation were the same as the observation group. Before and after treatment, blood pressure and syndrome score were observed and the short-term and long-term efficacies were assessed in the two groups.</p><p><b>RESULTS</b>After treatment, the systolic pressure and diastolic pressure were reduced as compared with those before treatment in the two groups (all P<0.001), and the differences were significant in comparison between the observation group and the control group (P<0.01, P<0.05). The syndrome score after treatment was reduced as compared with that before treatment in the two groups (both P<0.001) and the difference was not significant in comparison between the two groups (P>0.05). In follow-up of the effective cases and remarkably effective cases in 3 months, the total effective rate in the observation group was 89.3% (25/28) and was 60.0% (15/25) in the control group, indicating the significant difference in comparison (P<0.05).</p><p><b>CONCLUSION</b>Acupuncture combined with anti-hypertensive plaster is the effective method in prevention and treatment of borderline hypertension. Regarding the long-term efficacy, this combined therapy achieves the superior improvement in the symptoms and physical signs of the patients of borderline hypertension as compared with simple acupuncture.</p>

Assessment on depressive status and the therapeutic effects of highly active antiretroviral therapy among anti-HIV-1(+) population / 中华流行病学杂志

<p><b>OBJECTIVE</b>To assess the depressive status and its influence on Chinese HIV-1(+) population, and how it was influenced by highly active antiretroviral therapy (HAART) and the CD4(+) T cell count.</p><p><b>METHODS</b>Anti-HIV-1(+) patients (age between 18 and 65 years old) who had met the criteria to commence the anti-HIV treatment but had not yet started, were selected from the Beijing Ditan Hospital between March 2011 and June 2012. BDI-II (Beck Depression Inventory) and a self-designed questionnaire were used to evaluate the baseline and the status of 48 weeks post the HAART treatment. Statistically, t test and the Wilcoxon rank sum test were used to compare the BDI scores under different conditions and before/after the HAART.</p><p><b>RESULTS</b>(1) Of 100 subjects: male to female ratio was 99:1; the average age was 31.37 ± 5.58 years; the average education background was of 13.13 ± 3.51 years; the unemployed percentage was 4%; time before being identified as anti-HIV-1(+) was 5.0 (1.0 - 21.0) months; the percentage being infected through homosexual contact was 83%. The baseline BDI score was 6.0 (3 - 10.25). (2) There was no significant difference (P > 0.05) in BDI score between those subjects having had education less or more than 12 years; the BDI score of patients whose anti-HIV-1(+) was significantly higher (P < 0.05) among those discovered within the past 6 months than those more than 6 months. The BDI score of patients whose baseline CD4(+) T cell count below 200 cells/µl was significantly higher (P < 0.05) than those with baseline CD4(+) T cell count greater than 200 cells/µl. The CD4(+) T cell count was significantly high (P < 0.001) after 48 weeks of anti-viral treatment, but the BDI score was not significantly different (P > 0.05). There was no significant change (P > 0.05) in the proportion of patients with different degrees of BDI score before and after 48 weeks of antiviral treatment.</p><p><b>CONCLUSION</b>Depression in HIV patients was most overt in the first six months when they were aware of the infection. The degree of depression was more severe in patients with baseline CD4(+) T cell count less than 200 cells/µl with improvement of immunity after the HAART did not alleviate the level of depression.</p>

Molecular mechanism of HL-60 cell apoptosis induced by baicalin / 中国实验血液学杂志

This study was aimed to investigate the effect of baicalin on proliferation and apoptosis of HL-60 cells and its mechanism. Cell proliferation was assayed by using Cell Counting Kit-8. The morphological changes of HL-60 cells were examined by light microscopy and nucleolus morphological changes were observed by fluorescent microscopy after Hoechst 33342 staining. The early cell apoptosis was detected by using flow cytometry with Annexin V-FITC/PI double staining. The expression of caspase-3, caspase-9, Bcl-2 and Bax mRNA was detected by RT-PCR and Western blot assay was carried out to examine Bax, Bcl-2, caspase-8 and cleaved caspase-3 expression. The results showed that Baicalin inhibited the proliferation of HL-60 cells in a time- and concentration-dependent manner. HL-60 cells exhibited typical morphological features (for example, cell shrinkage, membrane blebbing and formation of apoptotic bodies). Cell apoptosis in early stage could be detected, the expression of caspase-3, caspase-9 and Bax mRNA was obviously up-regulated, while the Bcl-2 expression down-regulated, and accordingly Bcl-2/Bax ratio decreased. Such results were consistent with the expression of these proteins. In addition, the expression of cleaved caspase-8 protein was induced significantly after treated with baicalin. It is concluded that baicalin can significantly inhibit the proliferation of HL-60 cells and induce the apoptosis of HL-60 cells, which may occur through decreasing Bcl-2/Bax ratio by intrinsic pathway and through extrinsic pathway. It suggests that baicalin may be a promising drug for the therapy of acute myeloid leukemia.

Effects of hermap gene on p-STAT5 kinases in signal transduction pathway during erythroid differentiation / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the effects of hermap gene on kinases in erythroid signal transduction pathway and investigate the mechanism of hermap on erythroid differentiation.</p><p><b>METHODS</b>The K562 cells expressing hermap and hermap-siRNA respectively were established for up- and down-regulating the expression of hermap gene. These K562 cells were then induced by Ara-C to erythroid differentiation and analyzed at 0, 24, 48, 72 and 96 h, respectively, for cell morphology and biphenylamine staining positive cells, determination of CD235a, CD36, kinases p-STAT5, p-Akt, p-MAPK and p-c-JUN by FCM; and quantification of hermap gene and γ (Aγ,Gγ) globin gene by FQ-PCR.</p><p><b>RESULTS</b>With up-regulating hermap gene and inducing by Ara-C, K562 cells were changing to low ratio of nucleus to cytoplasm, cytoplasm colour from basophilic to pinkish or amethyst tinge, increase of number of biphenylamine positive cells and expression of CD235a, CD36, γ (Aγ,Gγ) globin gene, hermap gene and p-STAT5 from 0 to 96 h. At 0, 24, 48, 72 and 96 h of culture, the positive rates of p-STAT5 cells were detected of 0.46%, 4.54%, 20.01%, 23.65% and 33.08%, respectively. This results demonstrated that there was a positive correlation between expression of p-STAT5 and hermap gene expression (P < 0.05).</p><p><b>CONCLUSION</b>hermap gene can stimulate erythroid differentiation of Ara-C induced K562 cells mainly through JAK/STAT5 signal transduction pathway.</p>

Observation on immediate analgesic effect of acupuncture at Shiqizhui (EX-B 8) only or multi-acupoints in patients with dysmenorrhea: a randomized controlled trial / 中国针灸

<p><b>OBJECTIVE</b>To compare the different therapeutic effect between acupuncture at Shiqizhui (EX-B 8) only and multi acupoints on dysmenorrhea.</p><p><b>METHODS</b>Thirty eight cases were randomly divided into a single acupoint group and a multi-acupoints group, 19 cases in each group. The single acupoint group was treated by acupuncture at Shiqizhui (EX-B 8) only, and the multi-acupoints group by acupuncture at Shiqizhui (EX-B 8), Sanyinjiao (SP 6), Diji (SP 8), Ciliao (BL 32). They were all treated from the first day when sudden intense pain occurs, one time each day, for 3 days in each menstrual cycle, the treatment of three menstrual cycles. The therapeutic effect and Visual Analogue Scale (VAS) were compared and the score of general frequency and severity of dysmenorrhea by using Cox Menstrual Symptom Scale (CMSS) were evaluated.</p><p><b>RESULTS</b>The cured rate was 68.4% (13/19) and the effective rate was 31.6% (6/19) in the single acupoint group, being similar to 78.9% (15/19) and 21.1% (4/19) in the multi-acupoints group (P > 0.05). VAS and the scores of general frequency and severity of dysmenorrhea were all significantly decreased after treatment in both groups (all P < 0.001), with no significant difference between the two groups (all P > 0.05).</p><p><b>CONCLUSION</b>Acupuncture at Shiqizhui (EX-B 8) only can be as effective as selecting multi-acupoints to cure essential dysmenorrhea.</p>

Establishment of the cell line K562 with stable expression of hermap and hermap-siRNA / 中国实验血液学杂志

In order to establish K562 line with stable expressions of hermap and hermap-siRNA, amplified hermap and hermap-siRNA were cloned into pEGFP-c1 and pRNAT to acquire hermap-pEGFP-c1 and hermap-siRNA-pRNAT, respectively. These two plasmids were electrotransferred into K562 cells, then were followed by culturing with G418. The result showed that the transfer rate of hermap-pEGFP-c1-K562 and hermap-siRNA-pRNAT-K562 plasmids were 10.0% and 9.3%, respectively. After selective culture by G418, these two cell lines were still able to express GFP. It is concluded that the eukaryotic expression plasmids containing hermap and hermap-siRNA have been successfully constructed, and the cell lines of hermap-K562 and hermap-siRNA-K562 are established, definitely contributing to further functional investigation on HERMAP and its interaction with other proteins.

Discussion and analysis on point "Siguan" / 中国针灸

Point "Siguan" is seen in the Miraculous Pivot: Jiuzhen Shieryuan for the first time. From then on the doctors of every dynasties cited point "Siguan" and explained its content. But the authors hold that their explanations do not conform with the meaning in the Huangdi's Internal Classic. Therefore, the authors put forward their explanations for point "Siguan" and indicate that correct understanding of point "Siguan" is very important for treatment of diseases of five-zang organs.

Observation on therapeutic effect of warming acupuncture and moxibustion combined with Chinese drugs on ovarian cysts / 中国针灸

<p><b>OBJECTIVE</b>To observe the therapeutic effect of acupuncture and moxibustion combined with Chinese herbs on ovarian cysts.</p><p><b>METHODS</b>Forty-six cases were randomly divided into a treatment group of 26 cases and a control group of 20 cases. The treatment group were treated with warming acupuncture and moxibustion at Zigong (EX CA 1), Qihai (CV 6), Sanyinjiao (SP 6), etc. and oral administration of Chinese herbs Quyu Decoction, and the control group were treated with Quyu Decoction only. One month later, their therapeutic effects were observed.</p><p><b>RESULTS</b>The cured rate and the effective rate were 53.8% and 88.4% in the treatment group, and 15.0% and 85.0% in the control group, respectively, the cured rate in the treatment group being significantly better than that in the control group (P<0.01).</p><p><b>CONCLUSION</b>Warming acupuncture plus moxibustion combined with oral administration of Quyu Decoction can significantly increase the cured rate for ovarian cysts.</p>