Quality evaluation of Huocao based on UPLC fingerprint and multi-component content determination / 中国中药杂志

Huocao(a traditional Chinese herbal medicine) moxibustion is a characteristic technology in Yi medicine suitable for cold-dampness diseases. Huocao, as the moxibustion material, is confusedly used in clinical practice and little is known about its quality control. In this study, UPLC method was used to establish the chemical fingerprint of non-volatile components in Huocao, and the contents of eight phenolic acids such as chlorogenic acid were determined. Multivariate statistical analysis was performed to obtain the indicator components of Huocao for quality evaluation, and thus a comprehensive evaluation system for the quality of Huocao was built. The UPLC fingerprints of 49 batches of Huocao were established, and there were 20 common peaks, of which eight phenolic acids including neochlorogenic acid and chlorogenic acid were identified. Except for three batches of Huocao, the similarity of the other 46 batches was higher than 0.89, suggesting that the established fingerprint method could be used for quality control of the medicinal herb. The correlation coefficient between entropy weight score of the eight phenolic acids and comprehensive fingerprint score in Huocao was 0.875(P<0.01), which indicated that the eight phenolic acids could be used as indicator components for the quality evaluation of Huocao. Furthermore, in multivariate statistical analysis on the common peaks of fingerprint and the contents of the eight phenolic acids, chlorogenic acid, isochlorogenic acid A and isochlorogenic acid C were screened to be the indicator components. The results revealed that the proposed method achieved a simple and accurate quality control of Huocao based on UPLC fingerprint and multi-component content determination, which provided useful data for establishing the quality standard of Huocao.

Selection and validation of reference genes for quantitative real-time PCR analysis in Paeonia veitchii / 中国中药杂志

Paeonia veitchii and P. lactiflora are both original plants of the famous Chinese medicinal drug Paeoniae Radix Rubra in the Chinese Pharmacopoeia. They have important medicinal value and great potential in the flower market. The selection of stable and reliable reference genes is a necessary prerequisite for molecular research on P. veitchii. In this study, two reference genes, Actin and GAPDH, were selected as candidate genes from the transcriptome data of P. veitchii. The expression levels of the two candidate genes in different tissues(phloem, xylem, stem, leaf, petiole, and ovary) and different growth stages(bud stage, flowering stage, and dormant stage) of P. veitchii were detected using real-time fluorescence quantitative technology(qRT-PCR). Then, the stability of the expression of the two reference genes was comprehensively analyzed using geNorm, NormFinder, BestKeeper, ΔCT, and RefFinder. The results showed that the expression patterns of Actin and GAPDH were stable in different tissues and growth stages of P. veitchii. Furthermore, the expression levels of eight genes(Pv-TPS01, Pv-TPS02, Pv-CYP01, Pv-CYP02, Pv-CYP03, Pv-BAHD01, Pv-UGT01, and Pv-UGT02) in different tissues were further detected based on the transcriptome data of P. veitchii. The results showed that when Actin and GAPDH were used as reference genes, the expression trends of the eight genes in different tissues of P. veitchii were consistent, validating the reliability of Actin and GAPDH as reference genes for P. veitchii. In conclusion, this study finds that Actin and GAPDH can be used as reference genes for studying gene expression levels in different tissues and growth stages of P. veitchii.

Main components from cultivated and wild Nardostachyos Radix et Rhizoma by LC-MS and GC-MS / 中国中药杂志

In this study, ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and gas chromatography-mass spectrometry(GC-MS) were combined with non-targeted metabonomic analysis based on multivariate statistics analysis, and the content of five indicative components in nardosinone was determined and compared by UPLC. The main chemical components of Nardostachyos Radix et Rhizoma with imitative wild cultivation and wild Nardostachyos Radix et Rhizoma were comprehensively analyzed. The results of multivariate statistical analysis based on liquid chromatography-mass spectrometry(LC-MS) and GC-MS were consistent. G1 and G2 of the imitative wild cultivation group and G8-G19 of the wild group were clustered into category 1, while G7 of the wild group and G3-G6 of the imitative wild cultivation group were clustered into category 2. After removing the outlier data of G1, G2, and G7, G3-G6 of the imitative wild cultivation group were clustered into one category, and G8-G19 of the wild group were clustered into the other category. Twenty-six chemical components were identified according to the positive and negative ion modes detected by LC-MS. The content of five indicative components(VIP>1.5) was determined using UPLC, revealing that chlorogenic acid, isochlorogenic acid A, isochlorogenic acid C, linarin, nardosinone, and total content in the imitative wild cultivation group were 1.85, 1.52, 1.26, 0.90, 2.93, and 2.56 times those in the wild group, respectively. OPLS-DA based on GC-MS obtained 10 diffe-rential peaks. Among them, the relative content of α-humulene and aristolene in the imitative wild cultivation group were extremely significantly(P<0.01) and significantly(P<0.05) higher than that in the wild group, while the relative content of 7 components such as 5,6-epoxy-3-hydroxy-7-megastigmen-9-one, γ-eudesmol, and juniper camphor and 12-isopropyl-1,5,9-trimethyl-4,8,13-cyclotetrade-catriene-1,3-diol was extremely significantly(P<0.01) and significantly(P<0.05) lower than that in the wild group, respectively. Therefore, the main chemical components of the imitative wild cultivation group and wild group were basically the same. However, the content of non-volatile components in the imitative wild cultivation group was higher than that in the wild group, and the content of some volatile components was opposite. This study provides scientific data for the comprehensive evaluation of the quality of Nardostachyos Radix et Rhizoma with imitative wild cultivation and wild Nardostachyos Radix et Rhizoma.

Research progress on effects of high altitude hypoxia environment on antiepileptic drugs / 中国药理学通报

Hie high altitude hypoxic environment affects the pharmacokinetic process of rlnjgs by changing the body's gastrointestinal emptying rate, organ blood flow, drug plasma protein binding rate, dnjg metabolizing enzymes and transporter expression.Epilepsy is a brain disease that requires long-term medication.Most anti-epileptic drugs have a low therapeutic index and a narrow range of effective blood drug concentrations.'Ilierapeu- tic dnjg monitoring (TDM) is commonly used clinically to find the best individualized medication method for antiepileptic dnjgs.rI1iis article summarizes the commonly used anti-epileptic dnjgs and their treatment windows in clinical practice, and analyzes the influence of the pharmacokinetics of anti-epileptic dnjgs in the high altitude hypoxic environment, so as to provide reference for the clinical use of anti-epileptic drugs at high altitude.

Effects of altitude hypoxia on cerebral blood distribution of sodium valproate in mice / 中国药理学通报

Aim To investigate the effects of altitude hypoxia on serum sodium valproate eoncentration and eerebral blood distribution.Methods Male mice were divided into control group and plateau group.Each group was given sodium valproate orally and intrave¬nously, respectively.UFLC-MS/MS was used to deter¬mine the concentration of sodium valproate in plasma and brain, and Western blot was used to detect the ex¬pression of P-gp in BBB.Results Compared with the control group, the ratio of brain/blood drug concentra¬tion in plateau group was up-regulated by 44.0% , 57.9% , 176.8% and 184.5% at 10, 30, 60 and 120 min, respectively.The ratio of brain/blood drug con-centration increased by 33.9% , 50.6% and 125.6% at 60 min, 120 min and 240 min in plateau group, re¬spectively.Compared with the control group, the ex¬pression of P-gp protein in BBB of mice in altitude group was significantly down-regulated by 58.46% (P < 0.05 ).Conclusions Compared with the control group, the brain/blood drug concentration ratio of val¬proic acid increases in high altitude hypoxia environ¬ment.Meanwhile, it is found that P-gp expression lev-el decreased in the brain mierovessels of mice under high altitude hypoxia environment, and the cerebral and blood distribution of valproic acid in mic increases in high altitude hypoxia environment.

Quality control study of standard decoction of raw and honey processed Eriobotryae Folium based on HPLC fingerprint / 中草药

Objective: To establish HPLC fingerprint of Eriobotryae Folium standard decoction and compare quality difference between raw and honey processed Eriobotryae Folium standard decoction, which can provide a reference for its quality control. Methods: An HPLC-DAD method was utilized. The mobile phase was acetonitrile-0.4% phosphoric acid setting for gradient elution. The HPLC fingerprints of 20 batches of standard decoction of raw and honey processed Eriobotryae Folium were established. The contents of neochlorogenic acid, chlorogenic acid, and cryptochlorogenic acid were determined simultaneously. Similarity and cluster analysis were chosen to evaluate the quality of standard decoction of raw and honey processed Eriobotryae Folium. Results: Both of the fingerprint and the contents of three kinds of chlorogenic acids of Eriobotryae Folium standard decoction had significant difference before and after the Eriobotryae Folium being processed by honey. Two chromatographic peaks were increased newly in honey processed Eriobotryae Folium. The No.1 peak refers to component of 5-hydroxymethylfurfural. The average contents of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid in raw Eriobotryae Folium standard decoction were 4.300, 4.306, and 5.432 mg/g respectively. The result of contents showed a significantly decrease in honey processed Eriobotryae Folium standard decoction. Their contents were 3.295, 3.460, and 4.118 mg/g respectively. The reduction rate of them were 23.29%, 19.06%, and 23.92% respectively. Conclusion: The method is concise and durable. It could not only be utilized to evaluate the quality of standard decoction of Eriobotryae Folium before and after processed by honey, but also to identify the quality differences of them. The study could be used for quality control of standard decoction of raw and honey processed Eriobotryae Folium, identify the quality difference of them and also provide a reference for quality control of their preparations.

Grading standard of processed Curcumae Radix / 中国中药杂志

As one of the three pillars of Chinese medicine industry, traditional Chinese medicines prepared in ready-to-use forms are important raw materials for clinical medication and production of Chinese patent drugs. By considering the literature of Curcumae Radix, a multi-source Chinese herb and the situation of market investigation, the modern evaluation method based on traditional grading was introduced for comprehensive evaluation of the processed Curcumae Radix. The correlation between traditional grading method and modern evaluation index was explored to establish the grading standard of Curcumae Radix. According to the comprehensive evaluation, Curcumae Radix was divided into four grades: superior, first, second and third grades under the guidance of the theory of traditional Chinese medicine. This study provides a new idea for the grading of multi-source processed Chinese medicine, achieving high quality and good price, which is helpful to improve the clinical efficacy.

Determination of curcumin, desmethoxycurumin, and bisdesmethoxycurum in Curcuma longa by QAMS / 中草药

Objective: To establish a quality evaluation method for Curcuma longa, quantitative of multi-component with a single-marker (QAMS), to determine the contents of three curcuminoids at the same time. Methods: An HPLC method was used, and a Chromstar™ C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase was methanol-4% glacial acetic acid (48:52) at a flow of 1.0 mL/min. The detection wavelength was 422 nm and column temperature was 30℃. Curcumin was used as the internal reference substance. The relative correlation factors of curcumin, desmethoxycurumin, and bisdesmethoxycurumin to curcumin were calculated and evaluated by standard curve method and QAMS. Results: The QAMS method could be used to determine the contents of three curcuminoids at the same time, and with no significant difference between the results of standard curve method (RSD < 2.0%). Conclusion: QAMS method is simple, feasible, and reproducible, which could provide a reference for further research on completing the quality control of raw material and processed C. longa.

Expression of plateau adaptation gene of rat tissues after plain acute exposure to high altitude / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To detect the expression of the plateau adaptablity gene(EPAS1, EGLN1 and PPARα) and proteins(HIF-2, PHD2 and PPARα) in rats blood, heart, liver, lung and kidney tissue after the rats exposed to high altitude.</p><p><b>METHODS</b>The Wistar rats were randomly divided into plain group(Shanghai, 55 m), acute exposure to high altitude 3400 m group, acute exposure to high altitude 4300 m group. Blood and organs of rats were collected in 1, 3, 5 days after arrival. Real time PCR and ELISA were used to compare the expression of plateau adaptablity gene and related protein between plain group and high altitude exposure groups.</p><p><b>RESULTS</b>The count of red blood cells, hemoglobin and HCT in high altitude 4300 m were higher than those in plain group. Compared with plain group, the expression of EPAS1 gene in blood, heart, liver and kidney tissue of rats at high altitude increased obviously(all P<0.05); the expression of EGLN1 in the heart, liver, brain and kidney increased, and PPARα gene in the heart, liver and kidney increased(all P<0.05). Compared with plain group, the expression of HIF-2 protein increased significantly at high altitudes in the liver, brain and kidney tissues. PHD2 and PPARα increased in the heart, liver and kidney.</p><p><b>CONCLUSION</b>Plateau adaptive genes(EPAS1, EGLN1 and PPARα) and protein(HIF-2, PHD2 and PPARα) differed in different altitude and different organizations. They might be used as target markers of plateau hypoxia.</p>

Protective effect of SP600125 against liver cell injury in rats under repeated and sustained high +Gz exposure / 解放军医学杂志

Objective To explore the effect of JNK inhibitor SP600125 on expression of JNK/c-jun in liver cells of rats under repeated and sustained high +Gz exposure and its mechanism of the effect. Methods Eighteen inbred adult male Wistar rats were randomly divided into control group, +10Gz group and SP600125 group (n=6). The rats in +10Gz group and SP600125 group were fixed to the rotating arm of a centrifuge with head towards the axis. The increase rate of acceleration was 1G/s with a peak-time of 3 minutes, and the +Gz exposure was repeated 5 times with an interval of 30 minutes. SP600125 was given to rats of SP600125 group 30 minutes before the first centrifugation by intraperitoneal injection. All of the animals were sacrificed 30 minutes after centrifugation. Blood samples were collected from inferior vena cava to determine the plasma level of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The expression of c-jun mRNA was determined by quantitative real-time RT-PCR (qRT-PCR). The expressions of p-JNK, JNK, p-c-jun and c-jun protein were determined by Western blotting. The morphological change in the liver tissue was observed after HE staining. Results The plasma level of ALT and AST, expression level of c-jun mRNA and p-JNK, p-c-jun, c-jun protein in the liver tissue of SP600125 group were significantly higher than those of control group (P0.05). HE staining revealed disorganized hepatic cords, irregular liver cells, vacuolar changes, and marked edema of hepatocytes, and collapsed hepatic sinusoids in +10Gz group, but these changes were alleviated obviously in SP600125 group. Conclusion SP600125 could alleviate the liver cell injury in rats under repeated and sustained high +Gz exposure.

Analysis of complications and outcomes after pulmonary resection in patients aged 80 years or over with non-small cell lung cancer / 解放军医学杂志

Objective To explore the therapeutic effect and degree of safety in patients aged 80 years or over with non-small cell lung cancer undergoing radical resection. Methods A retrospective survey of 97 elder patients aged 80 years or over, in whom non-small cell lung cancer was pathologically confirmed after partial resection of the lung in Chinese PLA General Hospital from Jan. 2003 to Dec. 2012. The peri-operative features were evaluated, including gender, age, history of smoking, pulmonary function, surgical procedure, histopathologic type, pathologic stage, etc. The relation of the postoperative complications and mortality with the factors mentioned above was analyzed. Results Among all patients, the surgical procedure comprised 36 lobectomies, 31 segmentectomies, and 30 wedge resections. The histopathologic diagnosis showed there were adenocarcinoma in 51 patients, squamous cell carcinoma in 29, large cell carcinoma in 9, adenosquamous cell carcinoma in 6 and neuro-endocrine cell carcinomas in 2. The disease stage was determined as I A in 55 cases, I B in 33, IIA in 7, III A in 2. The post-operative complications (POC) occurred in 14 of 97 patients (14.4%), and the most common complication was cardiovascular complication (9 cases), followed by pulmonary complication (5 cases). Only two patients died to the complications, one of them was post-operative pneumonia and respiratory failure, and the other one was acute myocardial infarction. The survival rate of the 97 patients was 91.7%, 70.2% and 52.8% at 1, 3 and 5 years, respectively, and in the patients with stage I disease, the survival rate was 93.9%, 73.6% and 54.1%, respectively. Conclusion Advanced age is not a contraindication to radical pulmonary resection in patients over 80 years old suffering from early stage non-small cell lung cancer.

Chinese word processing in nonfluent aphasic patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Chinese nonfluent aphasic patients experience apparent speech production deficit, but it remains less known in which part of Chinese speech production this deficit occurs. The present study aimed to examine the ability of nonfluent aphasic patients in Chinese orthography, phonological and semantic processing via two experiments. Experiment I disclosed the general pattern of deficit of Chinese nonfluent aphasic patients in speech production. Experiment II tested whether this deficit occurs in orthography, phonological or semantic processing.</p><p><b>METHODS</b>The present study adopted neuropsychological testing methods to compare speech production and Chinese word processing between nonfluent aphasic patients (the patient group) and normal individuals (the control group). Character reading and word reading tasks were used to test speech production. Chinese radical decision, rhyme decision and semantic decision tasks were used to examine word processing. Reaction time and the correct answer rate were collected.</p><p><b>RESULTS</b>The patient group had a longer reaction time and was more prone to errors in both character reading and word reading tasks than was the control group. For the patient group, there was no difference between the reaction time of character reading and word reading, the error rate of the former was higher than the latter. In radical decision task the reaction time and error rate to the radical "mu " were higher in the patient group than in the control group. In the rhyme decision task, the reaction time and error rate to the rhyme "ang" were higher for the aphasic patients. In the semantic decision task the reaction time to characters in the category of animals was higher for the aphasic patients, yet the error rate was not significantly different between the two groups.</p><p><b>CONCLUSIONS</b>Nonfluent aphasic patients seemingly have decreased speed of speech production and an increased error rate. There is a deficit in phonological processing of aphasic patients while their semantic processing may remain intact.</p>

Culture Conditions Optimization of Magnetotactic Bacteria WM-1 / 现代生物医学进展

The culture of Magnetospirillum magneticum WM-1 depends on several control factors that have great effect on the magnetic cells concentration. Investigation into the optimal culture conditions needs a large number of experiments. So it is desirable to minimize the number of experiments and maximize the information gained from them. The orthogonal design of experiments and mathematical statistical method are considered as effective methods to optimize the culture condition of magnetotactic bacteria WM-1 for high magnetic cells concentration. The effects of the four factors, such as pH value of medium, oxygen concentration of gas phase in the serum bottle, C:C (mtartaric acid: msuccinic acid) ratio and NaNO3 concentration, are simultaneously investigated by only sixteen experiments through the orthogonal design L16(44) method. The optimal culture condition is obtained. At the optimal culture condition (pH 7.0, a oxygen concentration 4.0%, C: C (mtartaric acid: msuccinic acid) ratio 1:2 and NaNO3 100 mg l-1), the magnetic cells concentration is promoted to 6.5×107 cells ml-1, approximately 8.3% higher than that under the initial conditions. The pH value of medium is very important factor for magnetic cells concentration. It can be proved that the orthogonal design of experiment is of 90% confidence. The results from the hysteresis of WM-1 shows that Hc = 230 Oe, Ms = 0.9 emu/g dry wt. Cells,and Mr / Ms = 0.50.

Culture Conditions Optimization of Magnetotactic Bacteria WM-1 / 现代生物医学进展

The culture of Magnetospirillum magneticum WM-1 depends on several control factors that have great effect on the magnetic cells concentration. Investigation into the optimal culture conditions needs a large number of experiments. So it is desirable to minimize the number of experiments and maximize the information gained from them. The orthogonal design of experiments and mathematical statistical method are considered as effective methods to optimize the culture condition of magnetotactic bacteria WM-1 for high magnetic cells concentration. The effects of the four factors, such as pH value of medium, oxygen concentration of gas phase in the serum bottle, C:C (mtartaric acid: msuccinic acid) ratio and NaNO3 concentration, are simultaneously investigated by only sixteen experiments through the orthogonal design L16(44) method. The optimal culture condition is obtained. At the optimal culture condition (pH 7.0, a oxygen concentration 4.0%, C: C (mtartaric acid: msuccinic acid) ratio 1:2 and NaNO3 100 mg l-1), the magnetic cells concentration is promoted to 6.5×107 cells ml-1, approximately 8.3% higher than that under the initial conditions. The pH value of medium is very important factor for magnetic cells concentration. It can be proved that the orthogonal design of experiment is of 90% confidence. The results from the hysteresis of WM-1 shows that Hc = 230 Oe, Ms = 0.9 emu/g dry wt. Cells,and Mr / Ms = 0.50.