Objective:To study the qualitative identification and quantitative analysis of Paeoniae radix Rubra in Fufang Fuqing lotion. Methods:TLC was used to identify Paeoniae radix Rubra. The content of paeoniflorinl was determined by HPLC. The mobile phase was acetonitrile-0. 01% phosphonic acid (13 ∶87), and the detection wavelength was 230 nm,the flow rate was 1. 0ml·min-1, the column temperature was 40℃,and the sample size was 10μl. Results:The results of TLC showed that the relevant spots were clear without any interference from the negative sample. The calibration curve of paeoniflorinl was linear within the range of 0. 070-4. 500 μg (r=1. 000 0). The average recovery was 98. 36% with RSD of 2. 73%(n=6). Conclusion:The methods are accurate and quick in the qualitative identification and quantitative assay of the preparation, which can be used for the quality control of Fufang Fuqing lo-tion.

Objective:To optimize the processing technology of prepared Morinda officinalis based on nystose content. Methods:Orthogonal experiments and the nystose determination method in Chinese Pharmacopoeia (2010 edition) were used to optimize the pro-cessing technology. A Welch AQ-C18(250 mm ×4.6 mm,5 μm)chromatographic column was applied with methanol-water(3∶97)as the mobile phase, the column temperature was at 25℃, the flow rate was 0. 8 ml·min-1 , the injection volume was 10μl, and the drift tube temperature was at 40℃ with low temperature atomization. Results:The linearity of nystose was good within the range of 0. 214 8-0. 644 4 mg·ml-1 and the recovery was 99. 5%(RSD=3. 5%,n=6). Conclusion: On the basis of nystose content determination, the optimal processing technology of prepared Morinda officinalis is as follows:liquorice of 4%, stir fry time of 15min and the boiling pot temperature of 200℃.

Objective To evaluate the application of high-performance liquid chromatography (HPLC) in diagnosis and screening of thalassemia. Methods Automated HPLC was used to measure HbF and HbA2 in 100 genetically diagnosed thalas-semic patients and 35 normal children. The results were compared with those from traditional tests including alkali denaturation test and cellulose acetate electrophoresis. The diagnose accordance rates, sensitivity and specificity were compared. Results Seventy-fourβthalassemia, 64 were heterozygous with single mutations and 10 were compound heterozygous with double muta-tions. Twenty-sixαthalassemia, 25 were compound mutations and one was heterozygous with single mutation. The HbF percent-age from HPLC was higher than that from alkali denaturation tests in either thalassemia or normal children (P<0.01). HbF level from HPLC inα-thalassemia was signiifcantly different from that in the normal children (P=0.011). The percentage of HbA2 from HPLC was higher than that from cellulose acetate electrophoresis (P=0.010). HbA2 in the single heterozygousβ-thalassemia were twice higher than that in the double heterozygous mutatedβ-thalassemia (P<0.01). The combination of HbF-HbA2 (≥4.0%) from HPLC with MCV (<80 lf) and MCH (<27 pg) had high accordance rates (99.3%), sensitivity (99.0%) and speciifcity (100.0%) in diagnosis of thalassemia. Conclusions When the results of HPLC are combined with MCV and MCH, it can be applied to the diagnosis of thalassemia with high speciifcity, high sensitivity and has high diagnostic accordance rate with genetic results. HPLC can be an ideal approach to screenβthalassemia.

Objective To observe expression levels of N-methyl-D-aspartate (NMDA) receptor and cholecystokinin (CCK) in the hippocampus and spinal cord in morphine withdrawal or tolerance mice treated by electroacupuncture or catgut embedding and explore the difference between the regulating effects of electroacupuncture and catgut embedding on morphine withdrawal and tolerance.Methods Fifty-six male C57BL/6J mice were randomly allocated to withdrawal control, withdrawal model, withdrawal catgut embedding and withdrawal electroacupuncture groups, and tolerance control, tolerance model, tolerance catgut embedding and tolerance electroacupuncture groups, 7 mice in each group. A model of morphine withdrawal was made by subcutaneous injection of morphine hydrochloride using 7-day increasing addiction method. The withdrawal control group was injected with an equal volume of normal saline at the same time points. In the withdrawal electroacupuncture group, electroacupuncture at bilateral points Shenshu was performed using a Han’s acupoint nerve stimulation device (HANS-200) at 15 min after an injection of morphine hydrochloride. In the withdrawal catgut embedding group, 0.5 cm chromic catgut was embedded in bilateral points Shenshu at 15 min after an injection of morphine hydrochloride. Addiction was promoted by intraperitoneal injection of naloxone 4 mg/kg at 10 o’clock on the seventh day’s morning and Withdrawal reactions were observed in the mice. The score was recorded using the Ryuta Tomoji opioid withdrawal symptoms evaluation scale. NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by enzyme-linked immunosorbent assay (ELISA). A model of morphine tolerance was made by subcutaneous injection of morphine 10 mg/kg. The tolerance control group was injected with tolerance normal saline 10 ml/kg at the same time. In the tolerance catgut embedding group, catgut was embedded in point Shenshu at the first day after model making. In the tolerance electroacupuncture group, point Shenshu was given electroacupuncture at the first day after model making. After seven days of treatment, NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by ELISA.Results There were statistically significant differences in hippocampal NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There was a statistically significant difference in hippocampal NR2B expression between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the withdrawal catgut embedding or withdrawal electroacupuncture group and the withdrawal model group (P<0.05). There were statistically significant differences in spinal cord NR2A, NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There were statistically significant differences in spinal cord NR2A and NR2B expressions between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There were statistically significant differences in hippocampal NR2A, NR2B and CCK expressions between the tolerance model and tolerance control groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the tolerance catgut embedding and tolerance model groups (P<0.05). There was a statistically significant difference in hippocampal NR1 expression between the tolerance electroacupuncture group and the tolerance model or tolerance catgut embedding group (P<0.05). There was a statistically significant difference in spinal cord CCK expression between the tolerance catgut embedding or withdrawal electroacupuncture group and the tolerance model group (P<0.05).Conclusions Both catgut embedding and electroacupuncture at point Shenshu have a reducing effect on morphine tolerance and withdrawal. The therapeutic effect of electroacupuncture is better than that of catgut embedding.

The discovery of microRNAs (miRNAs) has introduced a new paradigm into gene regulatory systems. Since inception, computational methods have been an invaluable tool complementing experimental approaches, and many discoveries have been obtained through combination of experimental and computational approaches. The knowledge that has been accumulated about the principles of miRNAs and target recognition were reviewed. The currently available computational methodologies and software for prediction of miRNA and their target genes also have been discussed.

Objective To investigate the predictive value of early measurement of serum inflammatory mediators for infectious pancreatic necrosis secondary to severe acute pancreatitis (SAP).Methods A retrospective analysis was performed for the clinical data of 166 patients who were admitted to our hospital from January 2012 to January 2017.According to the presence or absence of secondary infectious pancreatic necrosis,the patients were divided into infection group with 58 patients and non-infection group with 108 patients.The serum levels of inflammatory factors were compared between the two groups.The receiver operating characteristic (ROC) curve was used for the analysis of indices with statistical significance.The independent samples t-test was used for comparison of continuous data between groups,and the chi -square test was used for comparison of categorical data between groups.Results Of all 166 SAP patients,58 experienced secondary infectious pancreatic necrosis,resulting in an incidence rate of 34.9％.Compared with the non-infection group,the infection group had significantly higher serum lipase,procalcitonin (PCT),C-reactive protein (CRP),and APACHE Ⅱ score (t =8.679,20.416,18.429,and 8.563,all P ＜0.05).The ROC curve analysis showed that serum lipase,PCT,CRP,and APACHE Ⅱ score had areas under the ROC curve of 0.647,0.877,0.823,and 0.655,respectively,with cut-off values of 612.5 U/L,7.5 ng/ml,226.5 mg/L,and 16.5 points,sensitivities of 68.5％,91.2％,86.8％,and 60.5％,and specificities of 59.3％,83.6％,80.1％,and 68.7％,respectively.Conclusion The abnormally elevated serum levels of CRP and PCT have a certain predictive value for infectious pancreatic necrosis secondary to SAP with convenient and fast operation,and therefore,it holds promise for clinical application.

OBJECTIVETo investigate the feasibility of using new tracheal prosthesis made of biomaterials to replace extensive circumferential tracheal defects in mongrel dogs.

METHODSThree types of tracheal prostheses were developed, whose basic skeleton of tubular mesh was knitted with polypropylene monofilament and poly (lactic-co-glycolic acid) fiber. The inner side of type-I tubular mesh was first coated with polyurethane solution and then with collagen. The exterior of type-I was then immobilized with collagen-hydroxyapatite composites. In contrast, the internal and external walls of type-II were coated with polyurethane solution, which produced a prosthesis similar to a nonporous one, while type-III was coated only with collagen solution. Surgical resection and replacement of a segment of the cervical trachea was performed in 16 adult mongrel dogs. The efficacy of the implanted prosthesis periodically evaluated postoperatively.

RESULTSIn group A, only one died from prosthetic dehiscence, another from anastomotic leakage, and the others had uneventful postoperative courses. The implanted prosthesis was completely incorporated with the recipient trachea, where different length of reepithelialization occurred on the luminal surface of the reconstructed trachea. Macroscopic examination showed scattered and different sizes of neo-ossification surrounding the implanted prosthesis. The prosthesis was roentgenopaque when exposed to routine X rays. In contrast, a relatively high number of complications occurred postoperatively in group B and C.

CONCLUSIONType-I tracheal prosthesis may be used effectively for long-segment circumferential tracheal replacement, and appears very promising for clinical application, with further improvements in promoting the epithelialization.

Animals ; Biocompatible Materials ; Collagen ; Dogs ; Female ; Male ; Polyglycolic Acid ; Polypropylenes ; Polyurethanes ; Prostheses and Implants ; adverse effects ; Prosthesis Design ; Prosthesis Implantation ; Trachea ; surgery

Background Asthma is a complex disease involving genetic and environment interactions.Atopy is a strong risk factor for asthma.The airway epithelium not only forms a physical barrier but also provides immune defense against harmful materials.To explore the effects of airway epithelium on asthma,we hypothesized that environmental injuries could act on bronchial epithelial cells and damage the physical barrier,which might facilitate allergens to stimulate immunoreactions and play an important role in the pathogenesis of asthma.Methods Thirty eight-week-old male Wistar rats were randomly divided into five groups with six rats in each group:control group,asthma group,ovalbumin (OVA)+OVA group,lipopolysaccharide (LPS) group and LPS+OVA group.In the control group,0.9％ saline was injected intraperitoneally on day 1.Fourteen days later,the rats were exposed to aerosolized 0.9％ saline.In the asthma group,the rats were sensitized with an injection of 10 mg of OVA,followed by an aerosolized 2％ OVA challenge14 days later.The OVA+OVA group was sensitized by an inhalation 2％ OVA,20 minutes a day,from day 1 to day 7,and then OVA challenged in the same way as the asthma group.In the LPS group,LPS (200 μl,1 μg/μl) was given by airway on day 1 and day 3,with a simultaneous aerosol inhalation of 2％ OVA for 20 minutes a day from day 1 to day 7.Fourteen days later,the rats were challenged with saline as in the control group.While in the LPS+OVA group,LPS (200 μl,1 μg/μl) was given by airway on day 1 and day 3,with a simultaneous aerosol inhalation of 2％ OVA for 20 minutes a day from day 1 to day 7.Fourteen days later,the rats were challenged with OVA as in the asthma group.The expression of interleukin (IL)-4,interferon-gamma (IFN-y) and thymic stromal lymphopoietin (TSLP) in the lungs was detected by reverse transcription polymerase chain reaction (RT-PCR) and the pulmonary pathological changes were also observed.The level of IL-4,IFN-γ and IgE in plasma was detected by enzyme-linked immunosorbent assay (ELISA).Bronchoalveolar lavage fluid (BALF) was collected to conduct differential cell counts.Flow cytometry analysis was also used to count Th1 and Th2 cells.Results The pathological changes in the LPS+OVA group were similar to the asthma group,while in other groups,the pathological changes were not obvious.The ratio of lymphocytes in BALF,IL-4/IFN-γ in plasma and the expression of the TSLP and IL-4 in the asthma and LPS+OVA groups were higher than in the control group and the OVA+OVA group (P ＜0.05).The level of IgE was higher in the asthma,LPS and LPS+OVA groups than in the control group and the OVA+OVA group (P ＜0.05).By flow cytometry analysis,the Th1/Th2 ratio was lower in the LPS+OVA and asthma groups than in other groups (P ＜0.05).Conclusions The experiment results show that the injury to the bronchial epithelial layer may be the initial event of allergic responses.This finding implies that a rational approach to therapeutics would be to increase the resistance of the airways to environmental injuries rather than concentrating on suppressing inflammation.

Objective To investigate the effect of single dose intraosseous injection of simvastatin on tumor vascular structure and function in murine breast cancer. Methods BALB/c mice and 4T1 murine breast cancer cells were used to establish a subcutaneous xenograft model. The mouse model of orthotopic breast cancer received intraosseous injection of a single dose of simvastatin (50 μg) or vehicle only. Frozen tumor tissue sections were prepared for co-immunostained with CD31 andα-SMA. Evans blue dye was injected into the tail vein to observe the vascular permeability. The expression level of HIF-1αwas detected by immunohistochemistry. Results Immunofluorescence dual staining showed that intraosseous injection of simvastatin increased the number of perivascular pericytes in the tumor vessel(P < 0. 05), Evans blue dye content showed that in vivo vessel permeability in the tumor tissue was significantly decreased(P < 0. 05), and the immunohistochemistry results showed that local hypoxic area was significantly improved. Conclusions Single dose intraosseous injection of simvastatin can promote the normalization of tumor vasculature by improving the coverage of pericytes.

Objective:To investigate the anticancer activity and mechanism of aconitine on cell proliferation,invasion and migration of hepatoma carcinoma cell(HCC).Methods:The effect of aconitine at different concentrations on proliferation was calculated by MTT assay.The effects of aconitine on invasion and migration of HCC were measured by Transwell and wound healing assay.Western blot was employed to detect the protein levels of P38MAPK signaling pathway-related proteins.Results:The concentrations of 5,10,20 μg/ml were selected according to the results of pre-experiment.Aconitine(10,20 μg/ml) inhibits proliferation and invasion of MHCC97 cells markedly after cells were treated with aconitine for 4 days.Treatment with aconitine down-regulated the ability of migration and decreased the ratio of p-P38/P38 and protein levels of p-MAPKAPK and p-HSP27.Conclusion:Aconitine inhibits prolif-eration,invasion and migration,and the mechanism may related with P38MAPK signaling pathway.