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.

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 .

Objective The effects of nemo-like kinase(NLK)in inflammation of central nervous system were explored. Methods An animal model with central nervous system inflammation disease induced by intracerebroventricular infusion of lipopolysaccharide was constructed. A model of neuronal apoptosis induced by glutamate in PC12 cells was constructed. A pcdna 3.1-NLK over-expression plasmid and a pSilencer 4. 1-CMV-NLK small interfering plasmid were also constructed. The expression and location of NLK were detected using western blot analysis, double immunofluorescent staining, and cell counting kit-8 assay. Results The protein level of NLK was reduced after induction of inflammation in the brain(t =2. 718-3. 106, all P ＜0. 01), and NLK was only expressed in both cortical and hippocampal neurons. At the cellular level, NLK expression was gradually reduced along with neuronal apoptosis induced by glutamate in PC12(t =4. 032, P ＜0. 01). Over-expression of NLK would reduce the apoptosis of neurons induced by glutamate(t =3. 930, P ＜ 0. 01). Conversely, interfering the expression of NLK would enhance neuronal apoptosis(t = 2. 845, P ＜ 0. 01). Conclusion NLK can protect neurons by inhibiting apoptosis in the process of central nervous system inflammation.

5-fluorouracil (5-Fu) as an antimetabolite drug,has been widely used in the treatment of various solid tumors.However, due to the heterogeneity of tumor and the impact of the individual, some patients still have drug resistance or serious lethal toxicity reaction.The text focuses on the research progress of pharmacogenomics and pharmacogenetics of some forecast elements which affect the efficacy and toxicity of 5-Fu,and discussed their guiding role on the choice of chemotherapy drugs.

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.

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.

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.