To summarize Professor GAO Shuzhong's clinical experience in treating idiopathic tinnitus with a combination of acupuncture and Chinese meteria medica. It is believed that idiopathic tinnitus is mostly caused by weak lungs and spleen， kidney essence deficiency， liver constraint transforming into fire， and binding constraint of heart qi. Treatment advocates the combination of acupuncture and Chinese meteria medica in clinical practice. Acupuncture treatment mainly focus on the method of opening the orifices by syndrome identification in combination with Ermen （TE 21）， Tinggong （SI 19）， Tinghui （GB 2）， Shenmai （BL 62） to regulate qi and blood， and supporting with Baihui （GV 20）， Yintang （EX-HN 3）， Taichong （LR 3）， and Yanglingquan （GB 34） to soothe the liver， resolve constraint， and calm the mind. Oral administration of Chinese medicinal prescription usually includes modified Yiqi Congming Decoction （益气聪明汤） and Tongqi Powder （通气散）， and the external administration of Chinese medicinal prescription can apply self-prescribed Wenqing Powder （温清散） to navel moxibustion.

Objective A simulation study was conducted to explore the effect and performance of g-computation-based joint mixed-effects model(JMM)on causal inference for controlling unmeasured confounders in longitudinal studies.Methods Longitudinal data including baseline and two follow-up visits were generated by computer simulations.The simulation scenarios included different sample sizes,the presence or absence of unmeasured confounders,and effects of unmeasured confounders.Causal effects were estimated using g-computation-based JMM,linear mixed-effects model,fixed effects model,and longitudinal target maximum likelihood estimation,respectively.Indicators including mean absolute deviation(MAD),standard error,root mean square error(RMSE),and 95%confidence interval coverage(95%CI coverage)were used to evaluate and compare the causal inference performance.Based on the physical examination cohort data of the menopausal women,four models were used to estimate the causal association between serum follicle-stimulating hormone(FSH)levels and lumbar bone density in menopausal women respectively,verifying the causal inference performance of models in the real longitudinal data.Results JMM had a better accuracy of causal inference with controlling unmeasured confounders.But its estimation stability was slightly worse.When strong unmeasured confounders existed,only JMM can accurately estimate the causal effect,and its precision and authenticity were better in scenarios with large sample sizes.Conclusion JMM can effectively control the unmeasured confounders and perform approximately unbiased causal estimation in longitudinal studies.

Objective:To identify the differentially expressed proteins that caused hippocampal damage after liver ischemia-reperfusion (I/R) in rats.Methods:Eighteen clean-grade healthy juvenile male Sprague-Dawley rats, aged 2 weeks, weighing 20-30 g, were divided into 2 groups ( n=9 each) using a random number table method: sham operation group (S group) and liver I/R group (IR group). A rat model of liver I/R injury was prepared by restoring perfusion after 1 h of liver ischemia. The rats were sacrificed after being anesthetized at day 3 of reperfusion, and the hippocampal tissue was isolated and analyzed to obtain gene expression profiles. Differentially expressed genes were identified using the R software, and further protein interaction networks were constructed through Cytoscape and Kyoto Encyclopedia Genes and Genomes pathway analysis to determine the differentially expressed proteins. Quantitative real-time polymerase chain reaction and Western blot were used for validation. Results:A total of 45 differentially expressed proteins were identified by the proteomic analysis of hippocampal tissues, including 36 significantly up-regulated proteins and 9 significantly down-regulated proteins. The proteins with significant expression related to injury were identified from the PPI network complex using the CytoHubBA plug-in cystscape: Ras-related C3 botulinum toxin substrate (RAC2), HRAS, phosphatidylinositol-3-kinase inhibitor phosphatase and tensin homologue (PTEN), and N-methyl-D-aspartate ionotropic glutamate receptor 2b (GRIN2b). The results of quantitative real-time polymerase chain reaction and Western blot showed that the expression of RAC2, HRAS, PTEN, and GRIN2b in the hippocampal tissue was significantly up-regulated in IR group compared with S group ( P<0.05). The results of Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that differentially expressed proteins were significantly enriched in the expression of PD-L1 and its checkpoint pathway, long-term potentiation, and regulation of the Wnt signaling pathway in cancer. Conclusions:The mechanism by which liver I/R induces hippocampal injury may be related to the up-regulation of the expression of RAC2, HRAS, PTEN and GRIN2b in rats.

Abstract: Objective　To analyze the molecular serotyping and drug resistance of foodborne Listeria monocytogenes (LM)isolates from the food monitoring network of Liaoning Province, and to provide a reference for identifying and tracing the outbreaks of foodborne diseases. Methods　The Listeria monocytogenes isolates were identified, biochemical typing was conducted by VITEK biochemical identification and PCR amplification techniques were applied for molecular serotyping of 67 strains of Listeria monocytogenes detected from 2 797 food samples collected from 15 monitoring sites across the province during 2021-2022. Verification was performed according to the instructions of the serodiagnosis manual. The sensitivity of the strains to ampicillin, penicillin, meropenem, cotrimoxazole, and erythromycin was determined by broth dilution method. Results　In food from Liaoning Province, the molecular serotypes of Listeria monocytogenes carried were identified as 1/2a(3a), 1/2b(3b, 7), 1/2c(3c), and 4b (4d, 4e), with the predominant serotypes being 1/2a accounting for 62.7% (42/67), and 1/2b accounting for 28.4% (19/67). Among them, the 4b strain was isolated in 3 strains, representing 4.5%. The dominant strains of Listeria monocytogenes in 2021 and 2022 in Liaoning Province were 1/2a(3a) and 1/2b(3b, 7), with the most diverse types carried by meat and meat products. Double resistance to cotrimoxazole and erythromycin accounted for 11.9% (8/67), while resistance to only one cotrimoxazole accounted for 9.0% (6/67). Conclusions　The presence of pathogenic serotypes 1/2a(3a), 1/2b(3b, 7), 1/2c(3c), and 4b (4d, 4e) in food in Liaoning Province and the emergence of strains with one or more drug resistances indicate the existence of food safety issues caused by Listeria monocytogenes, posing a potential risk of listeriosis. There is a need to optimize source tracing methods and strengthen the confirmation of Listeria monocytogenes outbreaks.

Objective To establish a high-performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS)method for determining the plasma concentration of olverembatinib in leukemia patients,apply it to clinical drug monitoring,and provide reliable basis for rational drug use in clinical practice.Methods Ponatinib-d8 was used as an internal standard,and methanol was used to precipitate plasma proteins and extract olverembatinib.The chromatographic column was Welch Ultimate XB-C18 cloumn(50 mmx4.6 mm,5 μm),with a column temperature of 60 ℃.The mobile phase consisted of an aqueous solution(containing 0.1％formic acid+2 mmol/L ammonium acetate)-methanol solution(containing 0.1％formic acid),with a flow rate of 0.8 mL/min and gradient elution.Electrospray positive ion mode was used,with multiple reaction monitoring scanning.The quantitative ion pair of olverembatinib was m/z 533.3→260.1,the qualitative ion pair was m/z 533.3→433.3,and the internal standard ion pair was m/z 541.1 →260.2.The plasma samples of 40 leukemia patients taking olverembatinib were monitored and analyzed for concentration,and IBM SPSS Statistics 27.0 and OriginPro 2021 softwares were used for statistical analysis of the results.Results The linear range of olverembatinib was 1-250 ng/mL(r=0.998 0),the lower limit of quantification was 1 ng/mL,the extraction recovery rate was 100.28％～101.27％,the intra-day precision RSD was 1.15％～3.87％,and the inter-day precision RSD was 2.32％～3.68％.Conclusion This method is easy to operate,highly specific and sensitive,and can be used to determine the blood concentration of olverembatinib in leukemia patients.

BACKGROUND:In recent years,computer-assisted navigation has gradually attracted attention in the medical field.Some scholars have reported that the clinical effects of computer navigation in total knee arthroplasty vary.To explore the clinical effect of the new generation of Brainlab Knee 3 navigation system,this study is conducted. OBJECTIVE:To analyze the application effect of computer navigation system(Brainlab Knee 3)combined with gap balance technology in total knee arthroplasty. METHODS:A total of 71 patients received total knee arthroplasty at the Department of Joint Surgery of Dalian Second People's Hospital from November 2020 to May 2021.In the navigation group,35 patients underwent total knee arthroplasty by computer navigation combined with gap balance.In the traditional group,36 patients underwent osteotomy with traditional surgical instruments.The difference between medial and lateral joint spaces,knee range of motion,hospital for special surgery knee score,hip-knee-ankle angle,deviation values of coronal femoral component angle,coronal tibial component angle,and sagittal tibial component angle were compared between the two groups. RESULTS AND CONCLUSION:(1)In the navigation group,the differences of intraoperative medial and lateral joint gap at knee extension 0,1,and 2 mm were found in 19,14,and 2 cases,respectively.The gap difference was 0 mm in 18 cases,1 mm in 15 cases and 2 mm in 2 cases at 90° degree knee flexion.In the traditional group,the gap difference at knee extention was 0 mm in 10 cases,1 mm in 20 cases,and 2 mm in 6 cases.The gap difference at 90 degree knee flexion was 0 mm in 10 cases,1 mm in 15 cases and 2 mm in 8 cases.(2)The operation time in the navigation group was longer than that in the traditional group(P<0.05),and two patients in the navigation group developed lower limb intermuscular vein thrombosis after surgery,and none of the patients in the traditional group had complications.(3)The range of motion between the two groups increased significantly at 6 and 12 months after surgery,and the range of motion in the navigation group was higher than that in the traditional group(P<0.05).(4)At 12 months after surgery,the hospital for special surgery knee scores of both groups increased significantly,and the hospital for special surgery knee score in the navigation group was higher than that in the traditional group(P<0.05).(5)At 6 months after surgery,the deviation values of hip-knee-ankle angle,coronal femoral component angle,coronal tibial component angle,and sagittal tibial component angle of navigation group were significantly lower than those in the traditional group(P<0.05).(6)The results showed that computer navigation technique combined with gap balance in total knee arthroplasty had advantages in postoperative range of motion,lower limb alignment and the accuracy of tibial prosthesis component position,and joint function recovered well.

OBJECTIVE To investigate the effects of 3 different primitives or the same primitives with different characters of Tibetan medicine Zuomoxing[Caragana changduenais Liou f. with red heartwood, Caragana jubata(Pall.) Poir. with brown and white heartwood] on rats with pattern of toxic heat-induced blood stasis. METHODS　Ninety SD rats were randomly divided into the blank group, model group, aspirin-positive group, Changdu low-dose group(CDD), Changdu high-dose group(CDG), whitewood of Guijian low-dose group(GJBD), whitewood of Guijian high-dose group(GJBG), brown wood of Guijian low-dose group(GJZD), Brown wood of Guijian high-dose group(GJZG). Models with heat toxicity and blood stasis pattern were established by intraabdominal injection of carrageenan combined with tail vein injection of lipopolysaccharide. The effects of each group on blood rheology, coagulation four indices and blood routine were determined, and the content of arachidonic acid(AA), IL-1β, IL-6, TNF-α and thromboxane B2(TXB2) were measured with ELISA. RESULTS ①Blood rheology: Compared with model group, CDD and CDG significantly decreased whole blood viscosity(WBV), reduction viscosity of whole blood(WBRV), erythrocyte rigidity index(HGX), erythrocyte deformability index(EDI), whole blood relative index(WBRI) (P<0.01), and increased plasma viscosity(P<0.01). GJZG and GJZD significantly decreased HGX(P<0.01 or P<0.05), and increased plasma viscosity(P<0.01). GJBG and GJBG significantly decreased WBHSV, WBHSRV, HGX, EDI, and whole blood high shear relative index(WBHSRI)(P<0.01). ②Coagulation four indices: Compared with model group, CDD significantly reduced the thrombin time(TT)(P<0.01). GJZG significantly reduced activated partial thromboplastin time(APTT) and TT(P<0.01 or P<0.05). GJBD significantly reduced prothrombin time(PT) and APTT (P<0.01 or P<0.05). ③Blood routine: Compared with model group, GJZD and GJBD significantly decreased the percentage of monocytes(P<0.01 or P<0.05). The number of large platelets in CDD significantly increased(P<0.05). CDG significantly increased the platelet number, platelet hematocrit, and large platelet number(P<0.01 or P<0.05), and tended which to be normal. ④Inflammatory factors: Compared with model group, the levels of TNF-α, IL-6, TXB2 were significantly increased in CDD and CDG(P<0.01 or P<0.05). The levels of IL-6 and TXB2 were significantly increased in GJZD and GJZG(P<0.01). GJBD was significantly increased TXB2(P<0.01), and GJBG was significantly increased IL-1β, IL-6, and TXB2(P<0.01), while decreased AA(P<0.05). CONCLUSION Zuomoxing with separate sources have different degrees of effects on rats with pattern of toxic heat-induced blood stasis, and have different degrees of effects on hemorheology, coagulation factors, blood routine and inflammatory mediators, and the degree and trend of effects are different with different doses. The effect of promoting blood circulation and removing blood stasis was generally manifested as Changdu > whitewood of Guijian > Brown wood of Guijian. The effect of promoting blood circulation and removing blood stasis may be the result of multiple pathways and mechanisms.

Objective To establish a high-performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS)method for determining the plasma concentration of methylprednisolone in patients with hematological diseases,and apply it to guide the clinical application.Methods Plasma samples were subjected to methanol-precipitated protein containing internal standard methylprednisolone-d3.The HPLC system was equipped with an Ultimate XB-C18(4.6mm×50mm,5 μm particle size),maintained at 60℃,and 5 μl of the supernatant was injected.Mobile phases consisting of 0.1%(v/v)formic acid(1∶1 000)and 2 mmol/L ammonium acetate in water(B)and 0.1%(v/v)formic acid(1∶1 000)in methanol at a flow rate of 0.8 ml/min was used.The electrospray ionization(ESI)source was operating in positive ion mode.Multiple reaction monitoring(MRM)was applied for the detection of the components:Methylprednisolone mass-to-charge ratio(m/z)375.4 → 339.4(qualitative ions),methylprednisolone m/z 375.4→357.3(quantitative ions),methylprednisolone-d3 m/z 378.2→360.3.The peak area ratio of methylprednisolone to methylprednisolone-d3 was used as the quantitative basis.The concentration of methylprednisolone in plasma was calculated and its performance was investigated.Results The linear range of methylprednisolone was 10～1 000ng/ml(r2=0.996 7),and the lower limit of quantification was 10 ng/ml.The RSDs of intra-day and inter-day precision results were less than 15%and the relative recovery ranged from 99.52%～104.79%.For methylprednisolone,the samples were stable at-20℃after three repeated freeze-thaw cycles.The prepared samples were stable at room temperature and 4℃for 24h(RSDs<15%).The steady-state blood drug concentrations of methylprednisolone in 16 patients were in the ranges of 1～258 ng/ml.Conclusion The HPLC-MS/MS method can accurately,rapidly and simply detect the concentration of methylprednisolone,and be suitable for clinical application.

Objective:To explore the effect of Xingnao Kaiqiao acupuncture treatment on brain network reorganization for the patients with stroke recovery, and therefore understand the neural mechanism underlying Xingnao Kaiqiao acupuncture treatment.Methods:Prospective case series study. Thirteen acute ischemia stroke patients were recruited from the Department of Neurology, Shanghai Minhang Hospital of Integrated Traditional Chinese and Western Medicine from Aug 2018 to Oct 2019. They were treated with Xingnao Kaiqiao acupuncture once a day for 10 consecutive days in addition to routine treatments, and received clinical assessments before treatment and 14 days after treatment onset. EEG signals were recorded during the first acupuncture treatment, from before inserting the needles (the baseline), during needle retention, to after removal of the needles. The brain network was constructed using phase locking index, and its clustering coefficient (CC), characteristic path length (PL) and small-worldness (S) were analyzed using one-way repeated ANOVA.Results:Compared with the baseline, the CC of delta-band network (sparsity=0.10: t=3.306, P=0.006; 0.12: t=2.909, P=0.013; 0.14: t=2.331, P=0.038) and the PL of delta-band (sparsity=0.12: t=3.236, P=0.007; 0.14: t=2.754, P=0.017, 0.18: t=2.878, P=0.014) and alpha-band (sparsity=0.10: t=2.432, P=0.032) networks were significantly decreased during the needle retention stage. Clinical assessments demonstrated a significant treatment efficacy of Xingnao Kaiqiao acupuncture, and its efficacy which was indicated by improved NIHSS score, was significantly correlated with the CC changes in the delta band network from baseline to needle retention. The correlation was strongest when the network sparsity was 0.12 ( r=0.78, P=0.002). Conclusion:Xingnao Kaiqiao acupuncture can regulate the brain network of stroke patients in real time, and this immediate regulation maybe associated with its treatment effect.

The end-of-life care education of medical students is related to the development of hospice care in the future. This paper comprehensively reviewed the setting up situations of end-of-life care education courses at home and abroad, as well as the status quo of courses' implementation, including teaching contents, teaching methods, assessment methods, teaching staff, teaching evaluations and effects. Based on these aspects, we have made some thoughts and suggestions, in order to provide reference for the development of end-of-life care education courses in medical colleges and universities in China.