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.

Carcinoma, Hepatocellular ; enzymology ; pathology ; Glutathione Peroxidase ; Humans ; Liver Neoplasms ; enzymology ; pathology ; Thioredoxin-Disulfide Reductase

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: Migraine is usually located in the course of Shaoyang channel, and the curative effect of treatment with methods of treating Shaoyang disease by dispersing and dispelling wind (DW) as well as relieving pain (RP) on migraine is prominent, but how about the effects of these two ways on thrombogenesis in vitro and the function of platelet aggregation (PA)?OBJECTIVE: To probe into the effect of treatment of Shaoyang disease and methods of DW and RP on thrombogenesis in vitro and function of PA throughthe experiments of thrombogenesis in vitro and function of PA in rats, and investigate the compatible significance of DS and DW as well as RP through formula compatible experiment.DESIGN: Completely randomized grouping design and controlled trial.SETTING: Department of Prescription, College of Basic Medical Sciences,Heilongjiang University of Chinese Medicine.MATERIALS: The experiment was conducted in the Laboratory of Department of Prescription, Heilongjiang University of Traditional Chinese Medicine from March to August 2000. Sixty healthy adult Wistar rats were randomly divided into 6 groups: Normal control group, Shaoyang treatment group (group Ⅰ ), DW and RP group (group Ⅱ ), high-dose and whole-prescription group (group Ⅲ), low-dose and whole-prescription group (group Ⅳ), and positive control group with 10 rats in each group.METHODS:① Positive control group:Gastric perfusion of 0.034 g/mL compound divaside slice suspension (made by Harbin Second Biochemical Pharmacy co., Ltd) was given to rats (0.39/kg); Group Ⅲ and Ⅳ (ingredients: Chaihu 20 g, huangqin 10 g, banxia 15 g, gancao 10 g, chuanqiong 20 g, tianma 15 g, xixin 5 g, quanxie 5 g, wugong 5 g): Gastric perfusion of 1.5, 0.75 g/mL of complete prescription was given to rats (17.40, 8.70 g/kg);Group Ⅰ (ingredients: Chaihu 20 g, huangqin 10 g, banxia 15 g, gancao 10 g) and group Ⅱ (ingredients: Chuanqiong 20 g, tianma 15 g, xixin 5 g, quanxie 5 g, wugong 5 g): Gastric perfusion of 0.75 g/mL agents of treatment with DS and DW wind antidyne was given to rats as 8.70 g/kg; Normal control group: Gastric perfusion of normal saline of the same volume wasgiven to rats.Intervention on rats of all groups lasted 12 days.② Experiment of thrombogenesis in vitro: The cephalic artery of one side was seperated 2 hours after the last time of administration. The distal part was deligated, the blood current in proximal part was blocked with bulldog clamp and the arterial cannula was inserted into the cephalic artery. Loosened the bulldog clamp, blood of 1.8 mL was collected and put into the rotary ring of thrombogensis meter to rotate for 15 minutes at 17 r/min,and then poured the thrombus, measured the length and humid weight of thromb. After that,dried the humid thromb and measured the dry weight.③ Determination of PA:The packing fraction (PF) in the 1st and 5th minutes as well as the maximal packing fraction (MPF) and assembling inhibition ratio of platelet-rich plasma within 5 minutes were recorded with test.MAIN OUTCOME MEASURES: Effect of treating Shaoyang disease nephelometry.④ Differences of measurement data were compared with tand methods of DW as well as RP on thrombogenesis in vitro and the function of PA.RESULTS: A total of 60 rats were involved in the analysis of results. ①The length, humid weight and dry weight of thrombogenesis in vitro in group Ⅲ and Ⅳ, positive control group and group Ⅱ were obviously lower than those in group Ⅰ (P ＜ 0.05). ② The PF in 1st, 5th minute and the maximal aggregathe normal control group (P ＜ 0.05-0.01); The humid weight and dry weight of thromb in group Ⅳ were significantly lower than those in tion (MA) of blood platelet in rats of each group were significantly lower than those in the normal control group (P ＜ 0.05-0.01), and the curative effect on rats in the positive control group, group Ⅲ and Ⅳ as well as group Ⅱ were better than that in group Ⅰ.CONCLUSION: There are no significant effects of dispersing thrombogenesis in vitro and inhibiting PA by only treating Shaoyang disease. The effect of combined prescription of DW and RP as well as treatment of Shaoyang disease are obviously enhanced.

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 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.

Intermittent hypoxia (IH) is a direct consequence of obstructive sleep apnea syndrome (OSAS), which results in left ventricular dysfunction , remodeling and myocardial cell injury .Understanding the mechanisms of OSAS is important for us to prevent and treat the cardiovascular complications in the clinical practices .We summarize the effects of IH on left ventricular function , and analyze the mechanisms at the cellular and molecular levels , including oxidative stress, ion exchangers and inflammation .In addition, the effects of endothelin-1 and hypercapnia on IH-induced cardiac injury are discussed .

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.

BACKGROUND:Increasing evidence has demonstrated that bone marrow mesenchymal stem cel (BMSC) transplantation can promote skin, liver and lung repair in animal models; andLeonurus sibiricus L. has the ability of promoting blood circulation and regulating menstruation.
OBJECTIVE: To investigate the therapeutic effect of BMSC transplantation with leonurine on intrauterine adhesions (IUA) in rabbits and the relevant mechanism of action.
METHODS: After modeled using dual injury method, rabbit models of IUA were randomly divided into five groups: sham-operated group (sham), model group (IUA), BMSC transplantation group, leonurine treatment group and combined treatment group (BMSCs+leonurine). Rabbits in the sham group were only given normal saline rinsing after hysterotomy, while those in the latter three groups were correspondingly given intrauterine BMSC transplantation or/and intragastric administration of 4 mg/kg leonurine for 14 days. Morphological changes of the endometrium were observed using hematoxylin-eosin staining, and expression levels of transforming growth factor β protein, Smad3 protein and interferon-γ mRNA were detected using immunohistochemical staining and real-time fluorescence quantitative PCR, respectively.
RESULTS AND CONCLUSION:Compared with the model group, the degree of IUA was al significantly improved in the other groups, especialy in the combined treatment group. Moreover, BMSC transplantation, leonurine treatment and their combined use al could inhibit IUA-induced increase of transforming growth factorβ and Smad3 protein expression and IUA-induced decrease of interferon-γ mRNA level. Importantly, al these alternations were much more pronounced in the combined treatment group. Our results show that the combined use of BMSC transplantation and leonurine treatment can exert a synergistic effect in the improvement of IUA through the transforming growth factor β/Smad3 pathway.