AIM:To establish HPLC method for making a assay of nucleosines in Ningxinbao Capsules(Cepha-(lospovium) sinensis Chen.sp.nov). METHODS:The assay was performed on a C_(18) column(4.6 mm?250 mm,5 ?m).The mobile phase was composed of phosphate buffer(pH 7.8-8.0)-acetonitrile(83∶17).The flow rate was at 1.0 mL/min.The detection wavelength was at 260 nm. RESULTS:The linear ranges of uridine,adenosine and inosine lay in the concentration range of 4.168-83.36,4.480-89.60,5.230-104.6 ?g/mL,respectively(r=0.999 9). The average recoveries were 0.4%,1.8%,2.5%,respectively. CONCLUSION:The method is simple,rapid.It is suitable for the assay of nucleosines in Ningxinbao Capsules.

Objective To develop and validate the real-time fluorescent quantitative PCR(Q-PCR)method for the detection of 8 murine viruses. Methods The specificity,sensitivity and precision of the Q-PCR method were verified by four laboratories,and the virus simulated contamination test and blind sample detection were carried out simultaneously,of which the detection results were compared. The Q-PCR method was used to detect 26 batches of monoclonal antibody cell lines for SARS-CoV-2 vaccine production and 15 batches of other products of murine origin. Results The Q-PCR method used for detecting 8 kinds of murine viruses had no cross reaction with the same family and genus or other murine viruses. Except the sensitivity of laboratory 2 to ectromelia virus(EctV/Mouse Pox,MPV)was 2 × 10~2copies/μL,the sensitivity of laboratory 2 to other 7 viruses and 3 other laboratories to 8 murine viruses was 2 × 10~1copies/μL. Except the inter-assay CV of the copy number of mouse adenovirus(MAdV)detected by laboratory 3 was 37. 58%,the intra-assay and inter-assay CVs of the Ct and copy number of other 7 viruses detected by laboratory 3 and those of 8 viruses detected by other 3 laboratories were all less than 25%.The sensitivity of virus simulated contamination test met the parameter requirements. The coincidence rate of blind sample detection results by 4 laboratories was 100%. All the 26 batches of monoclonal antibody cell lines for SARS-CoV-2 vaccine production and 15 batches of other murine derived products were negative for 8 murine viruses. Conclusion Q-PCR method for murine virus has good specificity,sensitivity and precision,and can be used for the detection of murine derived biological products.

Objective To explore the characteristics of attachment and the effects of childhood trauma,emotional autonomy and parental rearing style of anxiety disorders.Methods 38 outpatients and hospital patients in Anhui Mental Health Center with anxiety were recruited as the study group,and 39 age-,gender-,and educationmatched adult college students or medical staff were screened as the control group.The study group and control group were assessed with adult attachment questionnaire3.1 (AAQ3.1),parental bonding instrument (PBI),childhood trauma questionnaire 28item short form (CTQ-SF) and emotional autonomy scale (EAS).Results ①Higher levels of insecure adult attachment (84.2％) were reported in the anxiety disorders group compared with the normal controls(10.3％),and the attachment type of two groups had statistical significance(x2 =42.28,P＜0.01).② The attachment of anxiety disorders,emotional autonomy,childhood trauma and the way of parenting was related to different degree.Attachment,childhood trauma and parenting were significantly increased in the anxiety disorders group compared with the normal controls(P＜0.05).The mother attachment factors were significantly correlated with individualized (r=0.405,P＜0.405),mother love (r=0.358,P＜0.358),mother control (r=0.467,P＜0.467),father control (r=0.460,P＜0.01),emotional abuse (r=0.473,P＜0.473) and sexual abuse (r=0.494,P＜0.01).The father attachment factors were significantly correlated with individualized (r=0.520,P＜0.01),emotional abuse (r=0.339,P＜0.339) and sexual abuse (r=0.378,P＜0.05).And individualized,mother love,parental control,emotional and physical abuse could be employed to predict attachment factor.Conclusion The attachment pattern is insecure in the anxiety disorders.Emotional autonomy,childhood trauma and parenting have a significant influence on attachment patterns of anxiety disorders.

BACKGROUND:Tumor cels are resistant to chemotherapeutic drugs, and drug resistance is closely correlated with tumor stem cels. Therefore, how to kil tumor stem cels wil become the key to the treatment of oral squamous cel carcinoma. OBJECTIVE:To study the effect of 5-fluorouracil on biological characteristics of tongue squamous cel carcinoma Tca8113 cels. METHODS:Viability of Tca8113 cels treated with different concentrations of 5-fluorouracil was determined by cel counting kit-8, and the best drug concentration and time were screened for subsequent experiments. Tca8113 cels without 5-fluorouracil acted as control group. Then the cel cycle and percentage of the side population cels in Tca8113 cels were determined by flow cytometry. Scratch test was used to determine the migration ability of Tca8113 cels. RESULTS AND CONCLUSION:Results from cel counting kit-8 showed that 5-fluorouracil inhibited the viability of Tca8113 cels positively in a time- and dose-dependent manner. Tca8113 cels under intervention with 50 mg/L 5-fluorouracil for 48 hours showed lowest cel viability. Flow cytometry results showed that in the experimental group, G0/G1 phase cels increased significantly compared with the control group (P=0.01), S phase cels decreased significantly compared with the control group (P=0.244), and G2/M phase cels disappeared completely. After treatment with 5-fluorouracil, the percentage of side population cels was increased significantly (P=0.00). The scratch test showed that in the experimental group, the cels had better ability of wound healing than those in the control group. In conclusion, 5-fluorouracil can enrich the cancer stem cel population in Tca8113 cels.

Objective To explore the best humidifying methods of artificial airway by comparison of different humidifying approaches.Methods 124 patients were divided into the control group (60 cases) and the treatment group (64 cases).Two methods,continuous airway humidifying by the trace injection pump and continuous airway humidifying by oxygen atomization were adopted.The humidifying effect,tolerance rate of patients,the airway complications and nursing cost were compared between the two groups.Results The method of continuous airway humidifying by oxygen atomization had better effect,was easy to be tolerated,less complications and less cost.Conclusions Continuous airway humidifying by oxygen atomization is a safe,effective,low-cost method,it is easy to be tolerated by patients and is worthy of being popularized.

BACKGROUND:Studies have shown that apoptosis is closely related to the pathogenesis of steroid-induced femoral head necrosis. The release of cytochrome c plays a very important role in the process of apoptosis.
OBJECTIVE:To observe the effects of cytochrome c on early steroid-induced femoral head necrosis.
METHODS:Twenty-four healthy adult male New Zealand rabbits at 5 months old were randomly divided into model group and control group (n=12 per group). Models of early steroid-induced femoral head necrosis were established by intragluteal injection of hormone combined with ear vein injection of horse serum. In the control group, rabbits were given ear vein injection of the same amount of physiological saline. At 2, 4 and 6 weeks after model establishment, histopathological changes of bilateral femoral head were observed by optical microscope, and the ratio of empty lacuna was calculated. Apoptosis of osteocytes was determined by TUNEL assay, and apoptotic index was calculated. Immunohistochemical method was used to determine cytochrome c and to calculate cytochrome c-positive expression rate.
RESULTS AND CONCLUSION:(1) The ratio of empty lacuna and apoptotic index:The model of early steroid-induced femoral head necrosis was successful y established in the experiment. Compared with the control group, ratio of empty lacuna, apoptotic index and expression rate of cytochrome c in osteocytes were significantly higher in the model group (P<0.05 or P<0.01). (2) Correlation analysis:Ratio of empty lacuna was significantly positively associated with apoptotic index at various time points in the model group (r=0.856, P<0.01). Expression rate of cytochrome c was significantly positively associated with apoptotic index at various time points in the model group (r=0.824, P<0.01). (3) These findings confirm that cytochrome c-involved apoptosis of osteocytes may play an important role in steroid-induced femoral head necrosis. Expression rate of cytochrome c in osteocytes is remarkably positively associated with the occurrence of steroid-induced femoral head necrosis in rabbits.

Activation of HGF/Met has been complicated in tumorigenesis, invasion and metastasis of various tumors. We summarize current knowledge on their association with invasion of thyroid carcinoma. By discussing the expression of Met and its possible mechanism and analyzing the activation of Met with tumor motility, angiogenesis and inflammatory reaction, we conclude that the research of the possible role of HGF/Met in thyroid carcinoma will advance knowledge on the diagnosis and management of this disease,as well as tumor invasion.

0.05),but existed between grades Ⅰ-Ⅲ and grade Ⅳ(P0.05).Conclusion PIAS3 is expressed in human gliomas,closely correlated with the pathological grade and malignant intensity.PIAS3 may be involved in the development and progression of gliomas.

The determination of fraxetin A and fraxetin B in Cortex Fraxini have been carried out by HPLC on the stationary phase:octodecyl-silicohydride bonded silica gel column. The mobile phase was methanol-water(24:76). The detection wavelength was at 348nm. There was a good linear relationship in a concentration range of 16?g/ml~176?g/ml. The correlation coefficients were 0. 9995 and 0. 9994,respectively. The recovery of the added sample were 99. 1 ?1. 5% (n=5) for fraxetin A and 99.8?2. 5%(n=5) for fraxetin B. This method showed 0. 8% of within-day accuracy and 6. 2% of between-day accuracy,involving the advantages of simplicity,quickness and accuracy.

Objective: To establish the identification of protein compositions in Rhizoma Arisaematis through researches of its taxonomy, pharmacognostic anatomy and chemical compositions.Methods: The protein compositions in Rhizoma Arisaematis were identified by HPCE taking potassium phosphate boric acid buffer solution (pH=9.0) as an electrophoretic buffer. The electric sampling time was 10s, 10kV; Separating pressure 25kV; column temperature=28℃;?=205nm; integral determination sensitivety value=0.05; sensitivity=0.1 AUFS. Also, the raw plant identification, medical material characteristics and microscope identification were performed. Results: Combined with pharmacognostic and microscopic identification, experiments indicated that herbs of Arisaematis genus have similar HPCE chromatographic profile but different finger peaks. Condlusion: This study provides a better basis for the identification of Rhizoma Arisaematis.