OBJECTIVE:To estabilish a HPLC-MS method for determining enalapril in human plasma METHODS:Alprozalam was added into plasma sample as an internal standard,then supernate of the sample was extracted through solid-phase extration column,washed with methanol and detected by HPLC-MS method:column,ODS C18;mobile phase,methanol-0 01% formic acid(45∶55);flow rate,0 8ml/min;capillary voltage,3 81kV;cone voltage,39 0V The selected ion was determined by EST RESULTS:The calibration curve was linear within the range of 2 5～400ng/ml r=0 9 996,the recovery was 102 2%,RSDs of intra -day and inter-day were 4 0% and 5 4%,respectively CONCLUSION:The method is accurate and sensitive with no endogenous interference It can be applied to studying the pharmacokinetics and bioavailability of enalapril tablets in humans

Objective To explore the rule of traditional Chinese medicine in treating tinnitus based on literature and Logistic multiple regression analysis. Methods Articles about tinnitus treatment using traditional Chinese medicine were searched in several databases, i.e. CNKI (1984-2013), VIP (1989-2013), CBM (1990-2013), and PubMed (1984-2013), and medication frequency was analyzed. Then, models for tinnitus medication were established through metrological method and Logistic multiple regression. Results The common prescriptions with highest frequency usage were Liuwei Dihuang Pill, Longdan Xiegan Decoction, and Erlong Zuoci Pill. Corni Fructus, Psoraleae Fructus, and Lycii Fructus were commonly used for kidney-deficiency type of tinnitus;Astragali Radix, Atractylodis Macrocephalae Rhizoma, and Codonopsis Radix were commonly used for the spleen-qi-deficiency type of tinnitus;Gardeniae Fructus, Gentianae Radix et Rhizoma, and Cyperi Rhizoma were commonly used for the liver-qi-dysfunction type of tinnitus;Pinelliae Rhizoma, Aurantii Fructus Immaturus, and Stephaniae Tetrandrae Radix were commonly used for the phlegm-fire disturbance type of tinnitus;Forsythiae Fructus, Menthae Haplocalycis Herba, and Mori Cortex were commonly used for the wind-fire-invasion type of tinnitus. Conclusion Analysis of medication frequency and Logistic multiple regression analysis can provide evidence and reference for the treatment of tinnitus and syndrome differentiation.

Objective To analyze the occurrence of clinical adverse drug reaction (ADR) of tonic Chinese medicinal herbs. Methods Articles about ADR of tonic Chinese medicinal herbs reported from January 1990 to February 2014 were collected from databases of CNKI, VIP, CBM, Pubmed and Embase according to incorporation and exclusion standard. Metrology analysis was also conducted. Results Totally 378 effective articles showed tonic Chinese medicinal herbs most likely to induce ADR were as follows:individual Chinese medicinal herbs including Polygoni Multiflori Radix, Ginseng Radix et Rhizoma, Lycii Fructus and Astragali Radix;injection containing Shenmai Injection, Shengmai Injection and Acanthopanax Injection;other herbal preparation covering Polygoni Multiflori tablets. The most common ADR involved the damage on skin tissue, nervous system and digestive system, with prominent relevant clinical symptoms such as itching, skin rash, dizziness, unconsciousness, nausea, vomiting, and liver dysfunction. Conclusion Correct understanding adverse reaction of tonic Chinese medicinal herbs will be conducive to rational drug use and avoiding risk.

OBJECTIVETo explore the effects of electrical stimulation at acupoints in the distribution area of auricular vagus nerve combined with sound masking on auditory brainstem response (ABR) and contents of neurotransmitters of γ-aminobutyric acid (γ-GABA), 5-hydroxytryptamine (5-HT) and acetyl choline (Ach) in inferior colliculus of tinnitus rats.

METHODSTwenty-four male adult SD rats were randomized into a control group, a model group, a 7-d treatment group and a 15-d treatment group. Except the control group, rats in the remaining groups were treated with intraperitoneal injection of 10% salicylate sodium at a dose of 350 mg/kg to establish tinnitus model. Rats in the control group were treated with injection of 0.9% NaCl. Rats in the 7-d treatment group and 15-d treatment group were treated with electrical stimulation at "Shenmen (TF₄)" and "Yidan (CO₁₁)" in the distribution area of auricular vagus nerve combined with sound masking, once a day, for 7 days and 15 days. The SigGenRP software of TDT system was applied to provide voice for single ear and collect the signal, and the voice threshold of ABR was tested. The levels of γ-GABA, 5-HT and Ach in inferior colliculus of rats were detected by enzyme linked immunosorbent assay (ELISA) and compared.

RESULTSCompared with the model group, the threshold values of ABR in 12 kHz and 16 kHz voice stimulation in the 7-d treatment group were significantly lower all P < 0.05); the threshold values of ABR from 4 kHz to 28 kHz voice stimulation in the 15-d treatment group were signally reduced (P < 0.05, P < 0.01), which was more significant than those in the 7-d treatment group. The level of γ-GABA in the model group was significantly lower than that in the control group (P < 0.05), and that in the 15-d treatment group was apparently higher than that in the model group (P < 0.05). The level of 5-HT in the model group was markedly higher than that in the control group (P < 0.05), and that in the 7-d treatment group was lower than that in the model group (P < 0.05), while that in the 15-d treatment group was apparently higher than that in the model group (P < 0.05). The level of Ach in the model group was obviously; lower than that in the control group (P < 0.05), and that in the 7-d treatment group was higher than that in the model group (P < 0.05).

CONCLUSIONElectrical stimulation at auricular vagus nerve combined with sound masking) could regulate the threshold of ABR, especially in the 15-d treatment group. This may be ascribed to modulating the levels of neurotransmitter of γ-GABA, 5-HT and Ach in inferior colliculus.

Acupuncture Points ; Animals ; Brain Stem ; physiopathology ; Electric Stimulation ; Evoked Potentials, Auditory, Brain Stem ; Humans ; Inferior Colliculi ; physiopathology ; Male ; Neurotransmitter Agents ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serotonin ; metabolism ; Tinnitus ; physiopathology ; therapy ; Vagus Nerve ; physiopathology ; gamma-Aminobutyric Acid ; metabolism

OBJECTIVE: To explore changes of olfactory and gustatory functions in patients with olfactory dysfunction. METHOD: The 284 study subjects included 92 healthy volunteers, 92 with hyposmia and 100 with functional anosmia. Their olfactory and gustatory functions were examined using T&T olfactometer, olfactory event-related potentials (OERPs) and triple drop method, respectively. RESULT: The T&T results showed that the difference between patients with hyposmia and functional anosmia and healthy subjects had statistical significance. The OERPs results showed that patients with olfactory dysfunction had N1 and P2 waves of prolonged latency and reduced amplitude when compared to healthy subjects with the difference of statistical significance. When compared to healthy subjects, patients with olfactory dysfunction had clear hypogeusia and the difference had statistical significance. There was no significant difference between female and male groups. CONCLUSION It is suggested that the apparently concomitant hypogeusia is in patients with olfactory dysfunction. Decreased of olfactory and gustatory function exhibited little or no relationship with gender. Combined examinations of OERPs, T&T and triple drop method, could make it possible to reflect the overall chemical sensory functions in subjects systematically and provide scientific evidence for clinical diagnosis and treatment early.
Adolescent ; Adult ; Aged ; Ageusia ; diagnosis ; physiopathology ; Case-Control Studies ; Evoked Potentials ; Female ; Humans ; Male ; Middle Aged ; Olfaction Disorders ; diagnosis ; physiopathology ; Smell ; Taste ; Young Adult

Objective:To examine the correlation between DEPDC5 rs5998152 variants and the risk of hepatitis C virus (HCV) related liver diseases onset. Methods:Patients with chronic hepatic diseases diagnosed as HCV infection in Tianjin Second People′s Hospital from September 2016 to July 2017 were enrolled in the study and were divided into chronic hepatitis C (CHC) group, CHC related liver cirrhosis (LC) group and hepatocellular carcinoma (HCC) group. DEPDC5 rs5998152 was genotyped using the matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) assay and the case data were reviewed. T test, analysis of variance (ANOVA), and non-parametric test were used to perform the comparison of the quantitative data between groups according to normally distributed or not. Chi-square test was used to examine the different distribution of enumeration data between groups. Logistic regression analysis was employed to analyze the correlation between the genetic polymorphism and risk of LC and HCC. Results:A total of 147 patients were included in this study, with 55 in CHC group, 54 in LC group and 48 in HCC group. The levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), α-fetoprotein (AFP), total bilirubin and rate of hepatic encephalopathy were significantly higher in LC and HCC groups compared to CHC group( P<0.05). And the levels of AFP and total bilirubin were significantly higher in HCC group than LC group ( P<0.05). There was no significant difference among the three groups in terms of DEPDC5 rs5998152 genotype distribution ( P > 0.05). The frequency of the C allele at DEPDC5 rs5998152 was higher in LC and HCC subjects than in CHC patients ( P<0.05) and the Logistic analysis indicated that CHC individuals with C allele and TC+ CC genotypes showed higher risk of LC and HCC compared with those with T allele and TT genotype ( P<0.05). In addition, the difference of DEPDC5 rs5998152 allele frequency was not significant between LC and HCC groups and it was not correlated with risk of HCC for LC patients. Conclusions:DEPDC5 rs5998152 may be a risk factor of progression to LC and HCC in the Chinese Han patients with CHC.

Objective:Establishment of a new model of human primary colon cancer transplantation tumor in normal immune mice and to provide a reliable experimental animal model for studying the pathogenesis of colon cancer under normal immunity.Methods:Human colon cancer cells come from colon cancer patients who underwent surgery in the Affiliated Hospital of Jining Medical College in 2017. The mice in the cell control group were inoculated with phosphate buffered solution (PBS) containing colon cancer cells, the microcarrier control group was inoculated with PBS containing microcarrier 6, and the cell-microcarrier complex group was inoculated with the PBS containing colon cancer cell-microcarrier complex. The cells of each group were inoculated under the skin of the right axilla of mice by subcutaneous injection, and the time, size, tumor formation rate and pathological changes under microscope were recorded. The transplanted tumor tissue was immunohistochemically stained with the EnVisiion two-step method, and the tumor formation rate of the transplanted tumor was judged according to the proportion of positive cells in the visual field. The polymerase chain reaction (PCR) method was used to detect the expression of human-specific Alu sequence in mice tumor tissue.Results:After inoculation with tumor cells, the mice in the cell control group and the microcarrier control group did not die and did not form tumors; the mice in the cell-microcarrier complex group had palpable subcutaneous tumors in the right axillary subcutaneously on the 5th to 7th days after inoculation, and tumor formation rate is 67% (10/15), and the tumor volume can reach about 500 mm 3 2 to 3 weeks after vaccination. The immunohistochemistry results showed that CK20, CDX-2 and carcinoembryonic antigen were all positively expressed. The PCR results showed that the expression of human-specific Alu sequence can be detected in the transplanted tumor tissue of tumor-bearing mice. Conclusion:Human primary colon cancer cells used microcarrier 6 as a carrier to form tumors in normal immunized mice, and successfully established a new model of human colon cancer transplantation tumor in normal immune mice.

Objective:Establishment of a new model of human primary colon cancer transplantation tumor in normal immune mice and to provide a reliable experimental animal model for studying the pathogenesis of colon cancer under normal immunity.Methods:Human colon cancer cells come from colon cancer patients who underwent surgery in the Affiliated Hospital of Jining Medical College in 2017. The mice in the cell control group were inoculated with phosphate buffered solution (PBS) containing colon cancer cells, the microcarrier control group was inoculated with PBS containing microcarrier 6, and the cell-microcarrier complex group was inoculated with the PBS containing colon cancer cell-microcarrier complex. The cells of each group were inoculated under the skin of the right axilla of mice by subcutaneous injection, and the time, size, tumor formation rate and pathological changes under microscope were recorded. The transplanted tumor tissue was immunohistochemically stained with the EnVisiion two-step method, and the tumor formation rate of the transplanted tumor was judged according to the proportion of positive cells in the visual field. The polymerase chain reaction (PCR) method was used to detect the expression of human-specific Alu sequence in mice tumor tissue.Results:After inoculation with tumor cells, the mice in the cell control group and the microcarrier control group did not die and did not form tumors; the mice in the cell-microcarrier complex group had palpable subcutaneous tumors in the right axillary subcutaneously on the 5th to 7th days after inoculation, and tumor formation rate is 67% (10/15), and the tumor volume can reach about 500 mm 3 2 to 3 weeks after vaccination. The immunohistochemistry results showed that CK20, CDX-2 and carcinoembryonic antigen were all positively expressed. The PCR results showed that the expression of human-specific Alu sequence can be detected in the transplanted tumor tissue of tumor-bearing mice. Conclusion:Human primary colon cancer cells used microcarrier 6 as a carrier to form tumors in normal immunized mice, and successfully established a new model of human colon cancer transplantation tumor in normal immune mice.

OBJECTIVE： To provide reference for perfecting and improving the quality standard of Qiju dihuang oral liquid. METHODS： According to No. 0502 method stated in 2015 edition of Chinese Pharmacopeia （part Ⅳ）， TLC method was used to identify the fruit of Chinese wolfberry， Dendranthema morifolium and Paeonia suffruticosa in Qiju dihuang oral liquid. Using the fruit of C. wolfberry， D. morifolium and paeonol as control， the deployment systems were trichloromethane-ethyl acetate-formic acid （6 ∶ 1 ∶ 0.5， V/V/V）， trichloromethane-isopropanol-formic acid （10 ∶ 1 ∶ 0.5， V/V/V） and cyclohexane-ethyl acetate （3 ∶ 1， V/V）. The contents of morroniside， loganin and paeonol in Qiju dihuang oral liquid were determined by HPLC. The determination was performed on InertSustain C18 column with mobile phase consisted of acetonitrile-0.03％ phosphoric acid solution（gradient elution）at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm （morroniside and loganin） and 274 nm （paeonol）， and the column temperature was 40 ℃. The sample size was 10 μL. RESULES： In TLC of the fruit of C. wolfberry， D. morifolium and P. suffruticosa， same color spots were shown in the corresponding positions of reference substance/control chromatogram without interference from negative control. The linear ranges of morroniside， loganin and paeonol were 2.12-106.17， 1.91-95.63 and 4.78-239.16 μg/mL （R2＝0.999 9， 0.999 9， 0.999 8）， respectively. The limits of quantitation were 2.12， 1.91， 2.39 μg/mL； the limits of detection were 0.53， 0.48， 0.59 μg/mL， respectively； RSDs of precision， reproducibility and stability tests were lower than 2％ （n＝6）. The average recoveries were 98.27％， 97.06％ and 97.65％ RSD were 0.80％， 1.18％ and 1.36％ （n＝6）. RSDs of durability tests were all lower than 2％ （n＝3）. CONCLUSIONS： The established method is simple， specific and durable， and can provide reference for improving the quality standard of Qiju dihuang oral liquid.