ObjectiveTo investigate the role and mechanism of Lck/Yes-related novel protein tyrosine kinase （Lyn） on lipopolysaccharide （LPS）-induced M1-type polarization of macrophage. MethodsThe LentiCRISPR-V2 plasmid was digested with the restriction endonuclease BSMBI-V2， and the digested DNA fragments were recovered. The digested plasmid was ligated with Lyn-sgRNA using T4 ligase to generate the Lenti-Lyn-gRNA lentivirus. THP-1 cells were infected with the Lenti-Lyn-gRNA lentivirus to obtain a stable cell line with Lyn knockout， and a monoclonal THP-1 cell line with complete Lyn knockout （Lyn^⁻/⁻） was established subsequently. Wild-type Lyn （Lyn^WT） and Lyn^⁻/⁻ THP-1 cells were induced with 100 ng/mL phorbol myristate acetate （PMA） for 48 h to differentiate into M0 macrophages， which were further polarized into M1 macrophages by stimulation with 100 ng/mL LPS for 24 h. Quantitative real-time polymerase chain reaction （qPCR） was performed to detect the expression of M0 macrophage markers， including integrin αM （CD11b）， macrophage antigen （CD68）， and monocyte differentiation antigen （CD14）. The expression of Lyn in M1 macrophages differentiated from wild-type THP-1 cells （Lyn^WT-M1） was measured by qPCR， and the ratio of phosphorylated Lyn to total Lyn （P-Lyn/Lyn） in Lyn^WT-M1 cells was determined by Western blot. In M1 macrophages differentiated from Lyn-knockout THP-1 cells （Lyn^⁻/⁻-M1）， qPCR was used to detect the mRNA expression of inducible nitric oxide synthase （iNOS）， interleukin-6 （IL-6）， and chemokine （C-X-C motif） ligand 10 （CXCL-10）. Western blot was conducted to assess the protein expression of iNOS， as well as the protein levels of molecules related to the Janus kinase 1 （JAK1）-signal transducer and activator of transcription 1 （STAT1）signaling pathway， including JAK1， phosphorylated JAK1 （P-JAK1）， STAT1， and phosphorylated STAT1 （P-STAT1）. Additionally， the expression of the M1 macrophage marker cluster of differentiation 80 （CD80） was analyzed by flow cytometry. ResultsThe Lyn^-/- monoclonal cell line was successfully constructed. The expression of CD11b was significantly elevated in Lyn^-/- M0 macrophages， and the differentiation of M1 macrophages was successful. Knockdown of Lyn inhibited mRNA expression of iNOS， IL⁃6， CXCL⁃10， protein expression of iNOS and CD80 expression in M1 macrophages （P<0.05）. Western blot assay showed that Lyn knockdown inhibited protein expression of JAK1 and P-STAT1 （P<0.01）. ConclusionAfter CRISPR/Cas9-mediated Lyn knockout， the expression levels of JAK1 and P-STAT1， the key molecules in the JAK/STAT signaling pathway of M1 macrophages， are significantly downregulated； concomitantly， the expression of M1 macrophage-specific secretory factors （iNOS， IL⁃6， CXCL⁃10 mRNA） and CD80 is also downregulated， which may be achieved via targeted regulation of the JAK1/P-STAT1-mediated JAK/STAT signaling pathway.

Objective To observe the clinical effect of doing Huatuo five-animal play treating blood-stasis-type silent myocardial ischemia(SMI)and its influence on coronary hemodynamics.Methods Ninety patients diagnosed as silent myocardial ischemia were randomly divided into a five-animal play group(n=45,age:51.56±11.00 years,24 males and 21 females)and a regular exercise group(n=45,age:52.44±7.19 years old,24 males and 21 females)according to a random number table.The five-animal play group practised the traditional five-animal play,while the regular exercise group con-ducted daily 30-minute moderate-intensity treadmill aerobic exercise,five times a week,for 4 weeks.The changes in the number of abnormal ST-T segment occurrences,myocardial oxygen consumption(CMO),coronary ischemia threshold(CIT),and blood smoothness index(BSD)within 24-hour ambu-latory electrocardiogram before and after the intervention were observed in both groups,with the thera-peutic effect and hemodynamic characteristics of both groups evaluated.Results The average number of ST-T segment abnormalities decreased after intervention in both groups(P<0.001),with significantly greater improvement in the five-animal play group than the regular exercise group(P<0.001).More-over,the average CMO decreased significantly,while the average CMR and BSD increased significant-ly in both groups after intervention(P<0.05),with significantly greater improvement in the five-ani-mal play group than the other group.Meanwhile,the total effective rate in the five-animal play group was significantly higher than the regular exercise group(P<0.01).Conclusion Undergoing the five-ani-mal play and moderate intensity treadmill aerobic exercise both are effective in treating silent myocardi-al ischemia.However,the former therapy is superior to the latter in bettering CMO,CIT and BSD.

Objective To observe the clinical effect of doing Huatuo five-animal play treating blood-stasis-type silent myocardial ischemia(SMI)and its influence on coronary hemodynamics.Methods Ninety patients diagnosed as silent myocardial ischemia were randomly divided into a five-animal play group(n=45,age:51.56±11.00 years,24 males and 21 females)and a regular exercise group(n=45,age:52.44±7.19 years old,24 males and 21 females)according to a random number table.The five-animal play group practised the traditional five-animal play,while the regular exercise group con-ducted daily 30-minute moderate-intensity treadmill aerobic exercise,five times a week,for 4 weeks.The changes in the number of abnormal ST-T segment occurrences,myocardial oxygen consumption(CMO),coronary ischemia threshold(CIT),and blood smoothness index(BSD)within 24-hour ambu-latory electrocardiogram before and after the intervention were observed in both groups,with the thera-peutic effect and hemodynamic characteristics of both groups evaluated.Results The average number of ST-T segment abnormalities decreased after intervention in both groups(P<0.001),with significantly greater improvement in the five-animal play group than the regular exercise group(P<0.001).More-over,the average CMO decreased significantly,while the average CMR and BSD increased significant-ly in both groups after intervention(P<0.05),with significantly greater improvement in the five-ani-mal play group than the other group.Meanwhile,the total effective rate in the five-animal play group was significantly higher than the regular exercise group(P<0.01).Conclusion Undergoing the five-ani-mal play and moderate intensity treadmill aerobic exercise both are effective in treating silent myocardi-al ischemia.However,the former therapy is superior to the latter in bettering CMO,CIT and BSD.

Objective:To compare the clinical effects of ultrasound-guided intra-articular and coracohumeral ligament glucocorticoid injections versus ultrasound-guided intra-articular glucocorticoid injections alone on frozen shoulder in the elderly.Methods:A total of 44 patients with frozen shoulder admitted to the rehabilitation department of the Affiliated Hospital of Sichuan Nursing Vocational College from September 2019 to June 2020 were enrolled.They were randomly divided into the experimental group and the control group, with 22 patients in each group.The experimental group received two consecutive ultrasound-guided intra-articular injections and two coracohumeral ligament injections, while the control group received two consecutive ultrasound-guided intra-articular injections.The shoulder's passive range-of-motion(PROM), the 11-point numeric rating scale(NRS)as well as the Shoulder Pain and Disability Index(SPADI)were conducted before treatment, 1 week after the first treatment(1 week), 1 week after the second treatment(2 weeks)and 10 weeks after the second treatment(12 weeks).Results:There was no significant difference in baseline characteristics between the two groups(all P>0.05). Flexion, abduction, extension and external rotation increased whereas NRS and SPADI decreased after treatment, compared with pre-treatment in the experimental group( F=11.341, 20.965, 20.403, 32.470, 82.970 and 102.154, all P<0.05)and in the control group( F=7.012, 8.937, 23.265, 31.966, 52.500 and 41.356, all P<0.05). Compared with the control group, extension( t=0.365, 0.143 and 0.236, all P<0.05)and external rotation( t=0.205, 0.620 and 2.751, all P<0.05)increased at 1, 2 and 12 weeks after treatment.SPADI scores were lower in the experimental group than in the control group at 12 weeks after treatment( t=2.063, P=0.045). There was no significant difference in NRS, flexion or abduction between the two groups at any time point after treatment(all P>0.05). No serious adverse reactions occurred during treatment in either group. Conclusions:Both ultrasound-guided intra-articular injections plus coracohumeral ligament injections and ultrasound-guided intra-articular injections can effectively alleviate pain and improve function in elderly patients with frozen shoulder, but the former has better effects on extension.For patients with clear restricted extension and external rotation, ultrasound-guided intra-articular injections combined with coracohumeral ligament injections may achieve better clinical effects.

Objective: To investigate the current status of the ten-year implementation of the World Health Organization (WHO) surgical safety checklist (SSC) in China. Methods: A questionnaire was designed based on the three phases described by the checklist — the period before induction of anaesthesia, the period before skin incision, and the period before patient leaves operating room, taking into account some hotspots and disputes.The questionnaire was sent to the members of the Chinese-based online New Youth Anesthesia Forum through the WeChat platform.Answers were completed by mobile phones or desktop computers.Each WeChat ID number allowed only one answer for each individual participant. Results: A total of 3 943 members red the questionnaire invitation, of which 2 121 members completed the questionnaire with an overall completion rate of 53.79%.For checks completed before induction of anesthesia, the percentage of members who routinely practiced checks before induction of anesthesia was 93.35%, the percentage of members who completed each and every element of the checklist was 60.16%, and the percentage of members who selected the element of the checklist the surgeon was not involved in the check was 14.05%.For checks practiced before skin incision, the percentage of members who routinely completed checks before skin incision was 78.22%, the percentage of members who completed each and every element of the checklist was 51.91%, and the percentage of members who selected the element of the checklist surgeons and anesthesiologists routinely stated their own professional key information was 18.24% and 18.81%, respectively.For checks practiced before the patient leaved the operating room, the percentage of members who routinely completed checks before removing the patient from the operating room was 64.26%, and the percentage of members who completed each and every element of the checklist was 44.18%.The percentage of members who was really serious about practicing the checklist was 56.20%.The percentage of members who believed that surgeons should participated in checks practiced before induction of anesthesia was 81.47%.If the member himself or a member of his family needed a surgery, the percentage of members who hoped to implement the checklist was 98.35%.The percentage of members who believed that practicing WHO SSC could reduce the complications of surgery and improve the anesthetic safety of patients was 94.34%. Conclusion The implementing rate of checks practiced before induction of anesthesia is high, while the implementing rates of checks completed before skin incision and before patient leaves operating room are sequentially reduced in China.Although there are some problems with the implementation of WHO SSC, most respondents believe that implementing SSC can improve the anesthetic safety of patients undergoing surgery.

A simple, rapid, and sensitive gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the simultaneous determination of two fatty acids, methyl hexadecanoate (MH) and methyl stearate (MS), to allow the evaluation of packaging-drug compatibility. The two migrants were quantified in selective ion-monitoring (SIM) mode, with limits of detection (LOD) of 0.0030 μg/mL and 0.0121 μg/mL. Linear calibration curves for MH and MS were obtained in the concentration ranges of 0.1011–5.0570 μg/mL and 0.2015–10.0740 μg/mL, respectively. The developed method was successfully applied to estimate the safety of the injection of recombinant antitumor-antivirus protein (RAAP). The results showed that the possible maximum daily intake was 3.0 ng and 12.1 ng for MH and MS, re-spectively. As these values were both below the permitted daily exposure, the migrants can be con-sidered as having low safety risk and do not affect the quality of the injection.

A simple method was established for the determination of β-propiolactone (BPL) in human inactivated rabies vaccine by gas chromatography-mass spectrometry (GC-MS). The determination was performed on an Agilent HP-INNOWAX (30m × 0.32mm i.d., 0.25 μm) capillary column at the temperature of 80 °C. Electrospray ionization (ESI) was used by selective ion detection at m/z 42. The temperature for ESI source and inlet was set at 230 °C and 200 °C, respectively. Helium was used as the carrier gas at a flow rate of 25.1 mL/min. The total run time was 8 min. Acetonitrile and other components in the sample did not interfere with the determination of BPL. The results showed good linearity of BPL in the range of 0.50–10.01 μg/mL, with the limit of detection and the limit of quantification of 0.015 μg/mL and 0.050 μg/mL, respectively. Satisfactory precision was achieved for the current developed method. The method was applied to detect 6 batches of vaccine samples, and the results indicated that the target analyte BPL was present in three batches of unpurified samples, but was not detected in the purified samples, indicating the test samples were qualified. The established method was proved to be simple, versatile and sensitive, which can meet the requirements of quality control of BPL in human inactivated rabies vaccine.

OBJECTIVE:To screen and identify endophytic fungi from Schisandra chinensis with antioxidant activity. METH-ODS:The tissue isolation skill was used to isolate endophytic fungi from roots,leaves,stems and fruits of S. chinensis. And anti-oxidant activity of endophytic fungi were screened by DPPH radical scavenging assay and hydroxyl radical scavenging assay. The to-tal DNA were extracted;the 18S rDNA ITS were amplified and sequenced with primer ITS1 and ITS4;the results of sequencing were analyzed comparatively based on homology to confirm the classification of active strains. RESULTS:23 strains were isolated from S. chinensis. GSR-12,isolated from roots of S. chinensis,had strong antioxidant activities. The scavenging rate on DPPH and the hydroxyl radical were 87.96% and 82.31% respectively. GSR-12 strain was identified as Clonostachys rosea by analyzed com-paratively. CONCLUSIONS:1 strain of C. rosea,isolated from roots of S. chinensis,has strong antioxidant activity.

Objective:To explore the correlation between liver shear wave velocity(Vs) measured with acoustic radiation force impulse (ARFI) imaging technique and serum fibrosis indexes and its application in the diagnosis of liver cirrhosis and Child‐Pugh scoring for cirrhosis .Methods:The liver Vs value measured with ARFI technique of 150 patients with liver cirrhosis (ob‐servation group) and 50 healthy volunteers(control group) who visited Shanghai Dahua Hospital in Xuhui District during Jul 2013 and Jul 2014 were analyzed retrospectively .The 150 patients with liver cirrhosis were set as observation group and further divided into Class A(n=50) ,Class B(n=50) and Class C(n=50) according to Child‐Pugh scoring system .The 50 healthy vol‐unteers were set as control group .Serum fibrosis indexes testing ,including hyaluronic acid(HA) and laminin(LN) ,procollagenⅢ(PCⅢ) and collagenⅣ (Ⅳ‐C) ,were performed in all subjects .Differences between the observation group and the control group regarding indexes were compared by using independent sample t‐test .Pearson correlation analysis and receiver operator characteristic(ROC)curve analysis were conducted between Vs and four serum indexes in the observation group .Results:The values of Vs ,HA ,LN ,PCⅢ and Ⅳ‐C in the observation group were higher than those in the control group(P<0 .01) .Fur‐thermore ,the Vs value in the observation group demonstrated positive correlation with HA ,LN ,PCⅢ and Ⅳ‐C ,and the corre‐lation coefficients were 0 .672 ,0 .672 ,0 .765 and 0 .568 ,respectively(P<0 .01) .Area under ROC curve of the values of Vs and serum inedxes were 0 .85 0 .65 ,0 .71 ,0 .63 ,0 .73 in Class A ,0 .93 ,0 .83 ,0 .86 ,0 .84 ,0 .80 in Class B ,and 0 .99 ,0 .94 ,0 .85 , 0 .95 ,0 .94 in Class C ,respectively .Conclusions:Vs ,a non‐invasive marker for liver elasticity and stiffness ,is significantly correlated with serum fibrosis indexes .Its sensitivity and specificity are superior to those of four serum fibrosis indexes .It could provide qualitative analysis for evaluating the severity of liver cirrhosis ,so as to offer valuable evidence for further clinical treat‐ment .

Objective To investigate the expression of p73 gene and its protein and their relation with clinicopathologic features in nasopharyngeal carcinoma (NPC) tissues.Methods Expression of p73 mRNA and protein in 52 NPC and 25 normal nasopharyngeal tissues was detected by real-time fluorescent quantitative PCR and immunohistochemistry.Results Expression of p73 mRNA and protein was significantly higher in NPC than that in normal nasopharyngeal tissues (mRNA:73.1％ vs 24.0 ％,protein:71.2 ％ vs 36.0 ％),there were significant statistical differences between the two groups (P ＜ 0.05),and their expression was closely related to tumor invasion depth,degree of differentiation and clinical stage (P ＜ 0.05).Expression of p73 gene and protein was not closely related to age and gender (P ＞ 0.05).Conclusion Detection expression of p73 mRNA and its protein can be helpful in the diagnosis and prognostic evaluation in NPC.