Simultaneous rapid detection of ten stilbenes in serum of mice by UPLC-MS/MS / 中国中药杂志

Stilbenes is a class of natural polyphenols with 1,2-diphenylethylene as the skeleton structure which have structural and active diversity. However, there are fewer studies on their metabolic process, which limits the in-depth research and development of such components. An UPLC-MS/MS method simultaneously determining contents of ten stilbenes was firstly established in this study and applied to study the ten stilbenes of peony seed coats in the serum of C57 mice.Piceatannol was the internal standard, and methanol was used for protein precipitation, UPLC-MS/MS with negative ion mode was used for analysis, and the method was validated.The serum samples were collected and detected after mice being oral administered with 800 mg·kg~(-1) peony seed coat extracts for 8 weeks. The results showed that suffruticosol A, suffruticosol B, suffruticosol C, trans-ε-viniferin, cis-gnetin H, trans-suffruticosol D and trans-gnetin H were detected in serum samples, and the highest is suffruticosol A. The method is simple and quick with high specificity and sensitivity, and it is suitable for quantitative determination of ten stilbenes in the serum of mice.

Screening for potential biomarkers of traditional Chinese medicinechest impediment syndromesbased on plasma metabonomics / 中国药理学与毒理学杂志

OBJECTIVE To have a systematic pathomechanism view of three chest impediment-syndromes of Qi Deficiency and Blood Stasis syndrome(QDBS),Qi Stagnation and Blood Stasis syn-drome (QSBS), Cold Obstruction and Qi Stagnation syndrome(COQS) and further investigate the changed metabolome and related pathways for screening potential biomarkers in rat plasma. METHODS According to clinical pathogeny, three kinds of syndrome models were established to simulate the disease of chest impediment. Plasma metabonomics based on UPLC-Q-TOF/MS was applied in this research to detected small molecule metabolites for identifyingthe special potential biomarkers of three chest impediment syndromes, respectively. RESULTS Significant metabolic differences were observed between thecontrol group and three syndrome groups. Furthermore, three syndrome groups were distinguished clearly by pattern recognition method.The particular metabolites contributing most to the classification of three chest impediment syndromes were identified. In the QSBS group, the potential biomarkers could include 2-keto-glutaramic acid, L-methionine, L-homocysteic acid, octadecanamide, stearoylglycine,behenic acid,linoleylcarnitine,lysoPC(14:1(9Z)),indoxyl sulfate and cholic acid.In the COQS group, they could be aminoadipic acid, palmitic amide, oleamide, lysoPC(P-16:0), lysoPC(P-18:0), lysoPC(20:2(11Z,14Z)), 9-HETE and tauroursodeoxycholic acid. Moreover, 4-pyridoxic acid, L-palmi-toylcarnitine, lysoPC(20:0), lysoPC (22:5 (4Z,7Z,10Z,13Z,16Z)), 3- hydroxyhexadecanoic acid and arachidonic acid could be the potential biomarkers for the QDBS group. CONCLUSION Three chest impediment syndromes have their own potential biomarkers.Each special metabolite has its owndifferent metabolic pathway.Both metabolismof cysteine and methionine,and metabolism of alanine,aspartate and glutamate are the main pathways in regulation of metabolic disorders in QSBS syndrome. Lysine biosynthesis and degradation,fatty acid metabolism,and glycerophospholipid metabolism are the main pathways in regulation of metabolic disorders in COQS syndrome.Arachidonic acid metabolism, fatty acid metabolism,fatty acid elongation in mitochondria,and vitamin B6 metabolism are the main pathways in regulation of metabolic disorders in QDBS syndrome.These endogenous substances were indicated as the special potential biomarkers for three chest impediment syndromes and worth studying in depth.

Simultaneous determination of six Salvia miltiorrhiza gradients in rat plasma and brain by LC-MS/MS / 中国中药杂志

To develop a LC-MS/MS method for the determination of protocatechuic acid, protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone and tanshinone II(A) in rat plasma and brain. The plasma and brain samples were precipitated with ethyl acetate, then were separated on an Agilent eclipse plus-C18 column (2.1 mm x 50 mm, 3.5 microm) using acetonitrile (consisting of 0.1% formic acid) and water (consisting of 0.1% formic acid) as mobile phase in gradient elution mode. The mass spectrometer was operated under both positive and negative ion mode with the ESI source, and the detection was performed by MRM. The transition of 154.3/153.1 m/z for protocatechuic acid, 137.3/108 m/z for protocatechuic aldehyde, 493.0/295.2 m/z for Salvianolic acid A, 718.0/520.0 m/z for salvianolic acid B, 321.4/152.3 m/z for chloramphenicol, 297.4/254.3 m/z for cryptotanshinone, 295.5/249.3 m/z for tanshinone II(A) and 285.2/154.0 m/z for Diazepam. The calibration curves in the range of 0.625-1 000 microg x L(-1) for protocatechuic acid and protocatechuic aldehyde, 1.25-1 000 microg x L(-1) for salvianolic acid A, 2.5-1 000 microg x L(-1) for salvianolic acid B, 0.15-1 000 microg x L(-1) for cryptotanshinone, 0.625-1 000 microg x L(-1) for tanshinone II(A) are with good linearityin rat plasma and brain. The analysis method is sensitive, simple, and suitable enough to be applied in the pharmacokinetic study of the 6 main components. Animal testing gives the lgBB of the drugs and further studies of the 6 components cross the blood-brain barrier can be carried out.

Pharmacokinetics study on Danshensu in rats by brain microdialysis and automated blood technique / 中国中药杂志

To research the pharmacokinetic of Danshensu in brain via microdialysis method and automated blood technique. A microdialysis probe was inserted into the left lateral ventricle, and then dialysate samples and blood samples were continuously collected after iv Danshensu. LC-MS/MS was used to determinate for Danshensu in the dialysate samples. The in vivo recovery was used for the calibration of probe. WinNonlin was used for analyzing all pharmacokinetic data. Pharmacokinetic parameters of DSS in blood and in brain showed that Ke, t1/2,, AUC0-t, MRT were 0.04, 0.018 min(-1), 16.64, 58.76 min, 812.59, 51.19 min x mg x L(-1), 15.28, 79.97 min, respectively. The results were indicated that the study was successfully established LC-MS/MS detection method for Danshensu. Microdialysis combined with automated blood technique could better reflect the dynamic characteristics of Danshensu in the rat brain, and it provides a new perspective for pharmacokinetic study.

Advance in studies on chemical constitutions, pharmacological mechanism and pharmacokinetic profile of dalbergiae odoriferae lignum / 中国中药杂志

Dalbergiae Odoriferae Lignum is a traditional Chinese medicine (TCM) commonly used for promoting blood circulation, relieving pain and removing blood stasis. Volatile oil and flavonoid compounds are two main chemical constituents of Dalbergiae Odoriferae Lignum. Modern pharmacological studies show that Dalbergiae Odoriferae Lignum has many effects, such as relaxing blood, increasing blood flow of coronary, anti-oxidation, anti-inflammation and antitumor. Dalbergiae Odoriferae Lignum, as a characteristic TCM with the potential of further development, is generally compatible with other TCMs to treat cardio-cerebral vascular diseases. This article summarizes studies on chemical composition, pharmacological action, pharmacokinetic procfile in vivo and TCM compatibility in recent years, in order to provide references for further studies.

Clinical investigation on schistosomiasis after a 16-year-interruption program regarding its transmission, in Jiaxing region of Zhejiang province / 中华流行病学杂志

Objective To investigate the epidemic pattern,diagnosis and treatment of schistosomiasis after the 16-years intervention program on its transmissionin Jiaxing region of Zhejiang province.Methods Clinical data of schistosomiasis patients during the last 10 years and pathological specimens with deposited schistosomal eggs during the last 8 years were retrospectively analyzed.Results The total numbers of schistosomiasis patients admitted to hospital increased from 194 in 2001 to 960 in 2010,and from 78 to 266 with complications.Number of the ones with accompanied diseases increased from 116 to 694.All the numbers of the above said three groups showed an yearly increase.The hardest hit age of the patients was between 60 and 74.The number of specimens with deposited schistosomal eggs increased from 192 in 2003 to 298 in 2010.While the ratio of specimens with deposited schistosomal eggs to the total number of pathological specimens became slightly decreasing,the average age of patients increased,Eggs were mainly deposited on appendix,colon,rectum,stomach,liver,gallbladder and small intestine.Conclusion Schistosomiasis still hit the Jiaxing region,with the average age of patients increased.

Multiplex real-time PCR method for rapid detection of Marburg virus and Ebola virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Marburg virus and Ebola virus are acute infections with high case fatality rates. A rapid, sensitive detection method was established to detect Marburg virus and Ebola virus by multiplex real-time fluorescence quantitative PCR.</p><p><b>METHODS</b>Designing primers and Taqman probes from highly conserved sequences of Marburg virus and Ebola virus through whole genome sequences alignment, Taqman probes labeled by FAM and Texas Red, the sensitivity of the multiplex real-time quantitative PCR assay was optimized by evaluating the different concentrations of primers and Probes.</p><p><b>RESULTS</b>We have developed a real-time PCR method with the sensitivity of 30.5 copies/microl for Marburg virus positive plasmid and 28.6 copies/microl for Ebola virus positive plasmids, Japanese encephalitis virus, Yellow fever virus, Dengue virus were using to examine the specificity.</p><p><b>CONCLUSIONS</b>The Multiplex real-time PCR assays provide a sensitive, reliable and efficient method to detect Marburg virus and Ebola virus simultaneously.</p>

Study on canceration law of gastric mucosal dysplasia based on syndromes of Chinese medicine / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the syndrome evolution law of Chinese medicine (CM) in the patients with gastric mucosal dysplasia.</p><p><b>METHODS</b>Three hundred and twenty four gastric mucosal dysplasia patients with deficiency and excess correlation syndromes were enrolled by a multi-center collaboration for two years' clinical follow-up to detect the levels of tumor supplied group of factors (TSGF) and carcino-embryonic antigen (CEA).</p><p><b>RESULTS</b>Among the 324 cases, 29 cases turned cancer in the two years, and the canceration rate was 9.0%. The three syndromes with higher canceration rate were the damp-heat accumulating Wei syndrome concurring or combining with asthenia-cold in Pi and Wei syndrome for 16.7%; stagnation in Wei collaterals syndrome concurring or combining with asthenia of both qi and yin syndrome for 13.2%; stagnation of Gan and Wei qi syndrome concurring or combining with asthenia-cold in Pi and Wei syndrome for 8.0%, respectively. Among the three syndromes, the highest level of TSGF occurred in the former two syndromes. In the half year before carcinogenesis, the syndromes of the patients took on deficiency and excess concurrent syndromes, and the deficiency syndromes involving the qi and blood deficiency syndrome and the Shen deficiency syndrome accounting for 48.0%.</p><p><b>CONCLUSIONS</b>Gastric mucosal dyspalsia canceration syndromes took on the polymorphism of excess and deficiency concurrent syndromes and had the characteristics of deficiency syndromes involving qi and blood deficiency syndrome and Shen-yin-yang deficiency syndrome.</p>

Effect of gingerol on substance P and NK1 receptor expression in a vomiting model of mink / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Gingerol is the generic term for pungent constituents in ginger, which has been reported to be effective for inhibiting vomiting. We attempted to investigate the antiemetic effect of gingerol and its effective mechanism on substance P and NK(1) receptors in minks.</p><p><b>METHODS</b>The antiemetic effect of gingerol was investigated during a 6-hour observation on a vomiting model in minks induced by cisplatin, (7.5 mg/kg, intraperitoneal). The distribution of substance P and NK(1) receptors in the area postrema and ileum were measured by immunohistochemistry, and the expression of NK(1) receptor in the area postrema and ileum were measured by Western blotting.</p><p><b>RESULTS</b>The frequency of cisplatin induced retching and vomiting was significantly reduced by pretreatment with gingerol in a dose-dependent manner (P < 0.05). Substance P-immunoreactive was mainly situated in the mucosa and submucosa of the ileum as well as in the neurons of the area postrema. The immunoreactive production of NK(1) receptor was mainly situated in the muscular and submucosa of ileum and the neurons of area postrema, gingerol markedly suppressed the increased immunoreactivity of substance P and NK(1)1 receptor induced by cisplatin in a dose-dependent manner (P < 0.05), and exhibited effective inhibition on the increased expression levels of NK(1) receptor in both the ileum and area postrema dose-dependently (P < 0.05).</p><p><b>CONCLUSIONS</b>Gingerol has good activity against cisplatin-induced emesis in minks possibly by inhibiting central or peripheral increase of substance P and NK(1) receptors.</p>

Expression of p14(ARF) and E2F-1 in lung cancer in Gejiu and Xuanwei regions of Yunnan Province / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the correlations between p14(ARF) and E2F-1, and the role of their alterations in the tumorigenesis of the lung cancer in Gejiu and Xuanwei regions in Yunnan Province for providing the important experiment basis in revealing the molecular mechanism and looking for new markers for early diagnosis of lung cancer.</p><p><b>METHODS</b>The expression of p14(ARF) and E2F-1 was detected at theirs protein level by Immunohistochemistry S-P method in 30 specimens of lung cancer of Gejiu tin miners, 30 specimens of lung cancer of Xuanwei peasants and 20 specimens of normal lung tissue. E2F-1 mRNA was detected by ISH in 25 specimens of lung cancer of Gejiu tin miners, 25 specimens of lung cancer of Xuanwei peasants and 10 specimens of normal lung tissue. The positive signals were quantitatively analysed by HPIAS-100.</p><p><b>RESULTS</b>The positive unit (PU) of p14(ARF) and E2F-1 was 16.44 +/- 4.85 and 47.39 +/- 5.43 in Gejiu group, and 16.79 +/- 3.55 and 48.15 +/- 9.11 in Xuanwei group. Expression of p14(ARF) and E2F-1 protein in lung cancer of Gejiu and Xuanwei were statistically different compared with that in the normal lung (P < 0.01) respectively; The PU of E2F-1 mRNA was 48.58 +/- 7.75 in Gejiu group, and 49.41 +/- 8.53 in Xuanwei group, which was higher than that in normal tissue group. The differences were significant (P < 0.01). There was positive correlation between the expression of E2F-1 protein and E2F-1 mRNA in Gejiu group, Xuanwei group and normal group (P < 0.01, r = 0.833). The expression of p14(ARF) protein was significantly negatively correlated with the expression of E2F-1 protein (P < 0.01, r = -0.830).</p><p><b>CONCLUSION</b>There is the over-expression of E2F-1 gene and the deletion of p14(ARF) gene in the tumorigenesis of the lung cancer in Gejiu and Xuanwei regions in Yunnan Province. Over-expression of E2F-1 protein in lung cancer may be caused by enhanced transcription.</p>

Detection of enterovirus 71 and coxsackievirus A16 from children with hand, foot and mouth disease in Shanghai, 2002 / 中华儿科杂志

<p><b>OBJECTIVE</b>It was noticed that coxsackievirus A16 (CA16) and enterovirus 71 (EV71) were two major etiological agents of hand, foot and mouth disease (HFMD) in children. Recently there were several large outbreaks of HFMD in the Asia-Pacific region, and there was a propensity to cause severe complications or death in children under 5 years of age. The severe forms were associated with EV71 infection. Although epidemics of HFMD have been reported in the mainland of China, few reports about EV71 as the pathogen of HFMD epidemics are available. The present study was conducted to investigate the causal agent of an HFMD epidemic in children in Shanghai from April to June of 2002.</p><p><b>METHODS</b>Totally 102 specimens (including vesicle fluid, stool and throat swabs) were collected from 72 patients with HFMD. The specimens were inoculated into Vero and/or RD cells. At first all the isolates were respectively neutralized by the RIVM pools of enterovirus antiserum, the type-specific antisera to EV71 or to CA16. Secondly all untyped isolates were tested by RT-PCR assay with two specific primer pairs for VP1 genes of EV71 and CA16 respectively. The EV71 and CA16 were identified depending on the size of PCR products. Sequence analyses of VP1 genes of 9 virus strains were performed by the laboratory of China CDC.</p><p><b>RESULTS</b>Viruses were isolated from 91 specimens from 67 patients. Serotyping by neutralization failed for all the isolates. But the RT-PCR results indicated that the viruses isolated from 78 specimens from 58 patients were identified as positive for CA16 and the isolates from 13 specimens from 9 patients were identified as positive for EV71, the ratio between CA16 and EV71 was 6.4:1. The results of sequence analyses were consistent with those of PCR assay. Two EV71 strains isolated in this study belonged to a new lineage (C4) within genogroup C. One patient with EV71-associated HFMD had a complication of encephalitis with convulsion, shock, coma and dyspnea.</p><p><b>CONCLUSION</b>CA16 and EV71 were the primary causes of HFMD during the epidemic. It was the first report of EV71-associated severe encephalitis occurred in patients with HFMD in Shanghai.</p>

A clinical study of Suogudan granule in the treatment of rheumatoid arthritis / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the clinical efficacy of Suogudan Granule (SGDG) in the treatment of rheumatoid arthritis (RA).</p><p><b>METHODS</b>Ninety patients with RA were randomly divided into the treated group and the control group. The treated group was administered orally with SGDG 6 g each time, thrice a day, while the control group with the combined therapy of Fenbid Capsules 0.3 g each time, twice a day and Tripterygium tablet 20 mg each time, thrice a day. The treatment course for both groups was 6 weeks. The changes of clinical symptoms and signs, and laboratory indices such as erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), antistreptolysin O (ASO), routine examination of blood and urine, liver and kidney function, etc. before and after treatment were observed.</p><p><b>RESULTS</b>(1) The total effective rate in the treated group (88.0%) was obviously higher than that in the control group (67.5%) with significant difference (P < 0.05). (2) The improvement in arthralgia, joint swelling, time of morning stiffness, 15-meter walking, analgesia initiation and persistence in the treated group was better than that in the control group (P < 0.05, P < 0.01), but there was no obvious difference in improvement of joint tenderness, range of joint motion, grip strength, and initiating detumescence time (P > 0.05). (3) The improvement in ESR and RF in the treated group was better than that in the control group with significant difference (P < 0.05). The negative-conversion rate of ASO in the treated group was also higher than that in the control group (P < 0.01). (4) No evident abnormality in blood, urine, liver or kidney function was found in either group.</p><p><b>CONCLUSION</b>SGDG is effective and safe for the treatment of RA.</p>

Effect of combination of dizocilpine with general antiepileptic drugs on amygdala kindling models in rats / 药学学报

<p><b>AIM</b>To investigate the antiepileptic effect of dizocilpine (MK-801) on amygdala kindling models in rats and the effects of its combination with general antiepileptic drugs.</p><p><b>METHODS</b>To establish amygdala kindling models in rats and observe the effect of dizocilpine on kindling models and its combination with general antiepileptic drugs (phenobarbital, valproate and nicardipine) at ineffective dose. The influence of dizocilpine on convulsions induced by semicarbazide (SCZ) in mice were also observed.</p><p><b>RESULTS</b>Dizocilpine (0.1-0.25 mg.kg-1, i.p.) was shown to dose-dependently inhibit amygdala kindled seizure, shorten the after discharge duration (ADD) and reduce the Racine's stage (P < 0.01). The combination of dizocilpine with phenobarbital, valproate, nicardipine at ineffective dose shortened ADD or reduced Racine's stages (P < 0.01). Dizocilpine (0.1-0.25 mg.kg-1, i.p.) significantly prolonged the latency and reduced the rate of convulsions and death in mice.</p><p><b>CONCLUSION</b>Dizocilpine inhibits the seizure of the amygdala kindling and improve the antiepileptic activity of phenobarbital, valproate and nicardipine, indicating that these combination may provide a new approach for treating epilepsy.</p>