Objective: To investigate the effect of auricular therapy on sleep improvement and the GABAergic system pathway in a rat model of insomnia and to explore its possible mechanism. Methods: According to the random number table, 60 male SD rats were randomly divided into blank control, model, auricular point sticking, auricular bloodletting, and auricular bloodletting combined with sticking groups, with 12 rats per group. Insomnia was induced by intraperitoneal injection of p-chlorophenylalanine. After establishing the insomnia model, 36 rats were treated once a day with auricular point sticking or bloodletting for 5 consecutive days. After the intervention, the general condition and body weight of rats were observed; the righting reflex test was used to detect the sleep latency and duration; HE staining was used to observe the morphology of hypothalamic neuron cells; and an enzyme-linked immunosorbent assay was used to detect the GABA and glutamate content in rat serum. Immunohistochemistry(IHC) and real-time fluorescence quantitative PCR were used to detect GABA ARα1 and GABA ARγ2 protein and mRNA expression in the hypothalamus of rats, and Western blotting(WB) was used to detect GABA ARα1, GABA ARγ2, GAD65/67, GAT-1, and GABA-T protein expression in the hypothalamus of rats. Results: Compared with the blank control group, the model group had a lower body weight, a significantly shorter sleep duration (P<0.05), severe damage to the morphological structure of hypothalamic neurons with disordered cell arrangement, larger intercellular gaps, enlarged cell bodies, and a vacuolated appearance. All the intervention groups had significantly higher body weight and longer sleep duration than the model group (P<0.05). Compared with the other intervention groups, the auricular point sticking group had a longer sleep duration (P<0.05), and the hypothalamic neuron cells in all intervention groups improved, with the auricular point sticking group showing more apparent improvement. The model group had a lower GABA and higher glutamate contents, and GABA ARα1, GABA ARγ2, and GAD65/67 protein expression in the hypothalamus were lower than in the blank control group. In contrast, GAT-1 and GABA-T protein expression was higher, and GABA ARα1 and GABA ARγ2 mRNA expression was lower (P<0.05). The serum GABA content in the auricular point sticking and auricular bloodletting groups was higher, and the serum glutamate content in the auricular point sticking and auricular bloodletting combined sticking groups was lower than in the model group. GABA ARα1 mRNA expression in the hypothalamus of each intervention group was significantly increased, and GABA ARγ2 mRNA expression in the hypothalamus of the auricular point sticking and auricular bloodletting combined sticking groups increased. GABA ARα1(IHC, WB), GABA ARγ2(WB), and GAD65/67 protein expression in the hypothalamus of the auricular point sticking group increased, whereas GAT-1 and GABA-T protein expression decreased. GABA ARα1 and GABA ARγ2 protein expression(IHC, WB) in the hypothalamus of the auricular bloodletting group increased, whereas GABA-T protein expression decreased. GABA ARγ1(IHC) and GABA ARγ2(WB) protein expression in the hypothalamus of the auricular bloodletting combined sticking group increased, whereas GAT-1 and GABA-T protein expression decreased (P<0.05). Compared with in the inventation groups, the serum GABA content in the auricular point sticking group increased, the serum glutamate content decreased, GABA ARα1 and GABA ARγ2 mRNA expression in the hypothalamus increased, and GABA ARα1(IHC), GAD65/67 protein expression increased. In contrast, GABA-T protein expression decreased (P<0.05), and GABA ARγ2 protein expression(IHC) in the hypothalamus of the auricular bloodletting group increased (P<0.05). Conclusion Auricular therapy, particularly auricular point sticking, may have modulated the GABAergic system pathway by upregulating hypothalamic GABA ARα1, GABA ARγ2, and GAD65/67 protein expression while downregulating GAT-1 and GABA-T protein expression to alleviate symptoms in an insomnia rat model.

Objective To explore the application value of lymphocyte subsets combined with various cytokines in the disease progression of elderly patients with corona virus disease 2019(COVID-19).Methods From December 2022 to January 2023,146 elderly patients with COVID-19 diagnosed in the emergency ward of the Eighth Medical Center of PLA General Hospital were selected and divided into two groups according to the prognosis:127 cases in the COVID-19 survival group,19 cases in the COVID-19 death group.In addition,51 osteoporosis patients in geriatric medicine department were collected as control group.The proportion and absolute count of lymphocyte subsets(including T,B and NK cells),and 12 cytokines in plasma(including IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,TNF-α and IFN-γ)were compared between the control group and COVID-19 group,survival group and death group.The receiver operating characteristic(ROC)curve was used to evaluate its prognostic value in elderly patients with COVID-19 infection.Results Compared with the control group:① The proportion of NK cells in COVID-19 group was decreased,while the proportion of B cells was increased,and the differences were statistically significant(Z=-3.386,-4.140,all P<0.01).There was no significant difference in the proportion of T,CD8+T and CD4+T cells,and the differences were not statistically significant(Z=-1.244,-1.770,-0.951,all P>0.05).② The absolute numbers of T,CD8+T,CD4+T,NK and B cells in COVID-19 group were decreased,and the differences were statistically significant(Z=-9.418～-6.539,all P<0.01).③ The concentrations of IL-2,IL-6,IL-1β,IFN-γ,IL-8,IL-17,IL-12P70 and IL-10 in COVID-19 group were all increased,and the differences were statistically significant(Z=-8.851～-1.986,all P<0.05).There was no significant difference in the concentrations of IL-5,IFN-α,TNF-α and IL-4,and the differences were not statistically significant(Z=-0.460～-0.217,all P>0.05).Compared with the survival group:① There was no significant difference in the proportion of T,CD8+T,CD4+T,NK and B cells in the death group(Z=-1.873～-0.422,all P>0.05).② The absolute numbers of T,CD8+T and CD4+T cells in the death group were all decreased,and the differences were statistically significant(Z=-2.667,-2.287,-2.556,all P<0.05),while there was no significant difference in absolute numbers of NK and B cellsm and the differences were not statistically significant(Z=-1.934,-0.532,all P>0.05).③ The concentrations of IL-6,IFN-γ,IL-8,IL-17 and IL-10 in the death group were all increased,and the differences were not statistically significant(Z=-4.211～-2.655,all P<0.05),and there was no significant difference in the concentrations of IL-5,IFN-α,IL-2,IL-1β,IL-12p70,TNF-α and IL-4 the differences were not statistically significant(Z=-1.329～-0.279,all P>0.05).ROC curve analysis for the prognostic value of lymphocyte subsets combined with cytokines in elderly patients with COVID-19 showed that:the areas of total T cells,B cells and NK cells under ROC curve for predicting the prognosis of COVID-19 infection were 0.94,0.80 and 0.93,respectively.The areas of CD4+T cells and CD8+T cells under ROC curve for predicting the prognosis of COVID-19 infection were 0.93 and 0.90,respectively.The areas of IL-6,IFN-γ,IL-8,IL-17 and IL-10 in cytokines under the ROC curve for predicting the prognosis of COVID-19 infection were 0.91,0.71,0.87,0.74 and 0.90,respectively.However,the area of combined lymphocyte subsets and cytokines under ROC curve for predicting the prognosis of COVID-19 infection reached 0.99.Conclusion The immune status of elderly patients with COVID-19 was generally low.Evaluation of immune status has important clinical guidance significance in disease diagnosis,disease observation and prognosis.

BACKGROUND:Pro-kin balance system guidance has a relatively excellent rehabilitation effect on lower extremity proprioception and trunk control in stroke patients,but its effect on knee proprioception and balance function in patients after anterior cruciate ligament reconstruction has been less reported. OBJECTIVE:To investigate the effect of rehabilitation training guided by Pro-kin balance system on proprioception and balance function of the affected knee after anterior cruciate ligament reconstruction. METHODS:A total of 84 patients who underwent anterior cruciate ligament rupture reconstruction surgery were randomly divided into observation group and control group,with 42 patients in each group.The patients in the control group received routine rehabilitation intervention after surgery,and those in the observation group were given rehabilitation training based on the guidance of Pro-kin balance system.The training in each group lasted for 8 weeks.Lysholm score and International Knee Documentation Committee score were used to evaluate the change of knee joint function before and after the intervention.Average weight-bearing strength difference,trajectory error,swing value and 30°,45° and 60° passive angulation errors of the affected knee joint were used to evaluate the changes in the proprioception of the affected knee.The area and length of motion trajectory under open and closed eyes were used to evaluate the change of balance function.The satisfaction of patients in both groups with this rehabilitation training was investigated. RESULTS AND CONCLUSION:After training,Lysholm score and International Knee Documentation Committee score of patients in both groups were significantly higher than those before training(P<0.01),and the above scores in the observation group were significantly higher than those in the control group(P<0.01).After training,the average weight-bearing strength difference,trajectory error and swing value of the two groups were significantly lower than those before training(P<0.01),and the above scores in the observation group were significantly lower than those in the control group(P<0.01).After training,the passive angulation errors of 30°,45° and 60° of the affected knee joints in both groups were significantly lower than those before training(P<0.01),and those in the observation group were significantly lower than those in the control group(P<0.05,P<0.01).After training,the area and length of motion trajectory in both groups with eyes open were significantly smaller than those before training(P<0.01),and the above indicators in the observation group were significantly smaller than those in the control group(P<0.05,P<0.01).After training,the area and length of the movement track of the patients in both groups with eyes closed were significantly smaller than those before training(P<0.01),and the above indicators in the observation group were significantly smaller than those in the control group(P<0.01).The satisfaction of patients in the observation group was 95,which was significantly higher than 81%in the control group(P<0.05).To conclude,compared with the conventional rehabilitation training,the rehabilitation training based on Pro-kin balance system is more effective in improving the function,proprioception and balance function of the affected knee joints of patients undergoing anterior cruciate ligament rupture reconstruction,and the patients'satisfaction is higher.

Objective:To analyze the related factors that affect the occurrence of femoral head necrosis in patients with femoral neck fracture after surgery, and to build a nomogram predictive model.Methods:Using a retrospective study method, 180 patients with femoral neck fracture who were admitted to Hainan Provincial Hospital of Traditional Chinese Medicine from June 2019 to October 2021 were selected as the study objects. According to whether the patients had femoral head necrosis, they were divided into the occurrence group (75 cases) and the non occurrence group (105 cases). The receiver operating characteristic (ROC) curve was used to analyze the predictive value; Logistic regression was used to analyze the risk factors of postoperative femoral head necrosis; internal data were used to verify the clinical efficacy of nomogram model.Results:There was no significant difference in age, sex, body mass index and other general data between the two groups ( P>0.05); compared with the non occurrence group, Garden fracture in the occurrence group was mostly classified as type Ⅲ or Ⅳ, and more patients with preoperative traction, fracture displacement and unsatisfactory reduction quality, and the time from trauma to surgery was longer ( P>0.05). The area under curve of the time from trauma to surgery was 0.766 (95% CI 0.697 to 0.826), and the optimal cutoff value was 42 h; Garden fracture classification (type Ⅲ or Ⅳ), preoperative traction (yes), fracture displacement (yes), reduction quality (unsatisfactory) and time from trauma to surgery (>42 h) were the risk factors affecting the postoperative femoral head necrosis in patients with femoral neck fracture ( P<0.05). The risk of femoral head necrosis predicted by nomogram model was 0.802 (95% CI 0.692 to 0.873). The risk threshold of femoral head necrosis predicted by the model was >0.09, and nomogram model provides clinical net income. Conclusions:Garden fracture classification (type Ⅲ or Ⅳ), preoperative traction (yes), fracture displacement (yes), reduction quality (unsatisfactory) and time from trauma to surgery (>42 h) are the risk factors affecting the postoperative femoral head necrosis in patients with femoral neck fracture, and the nomogram model based on variables has a better predictive ability.

Objective:This study aims to establish an early warning diagnosis model for acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and to provide a simple, rapid, and accurate auxiliary diagnosis basis for clinical practice.Methods:The sample bank of subjects (patients admitted to the Eighth Medical Center of the PLA General Hospital from September 10, 2021, to July 25, 2023) was constructed, including the model establishment cohort [SCOPD group 49, 42 males and 7 females, (69.71±11.16) years old; AECOPD group 53, 49 males and 4 females, (72.60±10.19) years old] and the model validation cohort [SCOPD group 35, 28 males and 7 females, (69.97±10.40) years old; AECOPD group 35, 33 males and 2 females, (71.43±9.67) years old]. Fasting peripheral blood samples were collected, and the expression levels of IL-5, IL-17A, and IFN-α were detected by flow cytometry. Different expression levels were analyzed by Mann-Whitney U test. Binary logistic regression analysis was used to screen the related risk factors of COPD patients in acute exacerbation. The diagnostic efficacy of the model was evaluated by the receiver operating characteristic (ROC) curve.Results:The levels of IL-5 [1.64 (0.60, 2.86) pg/ml], IL-17A [1.42 (0.88, 2.29) pg/ml], and IFN-α [0.91 (0.59, 1.81) pg/ml] in the SCOPD group were significantly decreased compared with the AECOPD group IL-5 [4.68 (2.34, 9.40) pg/ml, Z=-5.033, P<0.001], IL-17A [2.33 (1.59, 4.62) pg/ml, Z=-3.919, P<0.001], IFN-α [2.83 (0.91, 3.75) pg/ml, Z=-4.127, P<0.01] in the cohort of model establishment. The results of binary logistic regression analysis between SCOPD and AECOPD groups showed that IL-5, IL-17A, and IFN-α were independent risk factors for acute exacerbation of patients with COPD ( P<0.05). And the regression equation is Y=-2.861+0.364×IL-5+0.385×IL-17A+0.445×IFN-α. The AUC value of IL-5, IL-17A, IFN-α and combined detection was 0.866 ( P<0.001). Compared to the SCOPD group and the AECOPD group in the cohort of model validation, the receiver operating characteristic (ROC) curve showed that the combined model of three (AUC=0.858, P<0.001) could be used to diagnose the AECOPD. And the Kappa value was 0.773( P<0.05). Conclusion:The combined detection of IL-5, IL-17A, and IFN-α has high diagnostic efficacy for patients with acute exacerbation of COPD. This method provides a new potential tool for the clinical diagnosis of AECOPD and has the value of further exploration and optimization, promotion, and application.

AIM: Previous studies have suggested that big stick design (BSD) method can only be used in clinical trials of two treatments with equal proportion, which has good statistical performance and has become the recommended choice of randomized methods. This study expands BSD method, so that it can be applied to three groups, and provides more randomized methods for clinical trials. METHODS: On the basis of BSD method used in two treatments with equal proportion, the derivation conditional allocation probability of BSD method used in three treatments with equal proportion was carried out. BSD method was compared with simple randomization (SR) method, permuted block design (PBD) method and block urn design (BUD) method by Monte-Carlo simulation in balance and randomness. RESULTS: In terms of balance, PBD method was the best, followed by BUD method, BSD method, and SR method was the worst. In terms of randomness, SR method was the best, followed by BSD method, BUD method and PBD method. The comprehensive performance showed that BSD method was better than BUD method, PBD method and SR method. CONCLUSION: The expanded BSD method used in three treatments with equal proportion has good comprehensive performance, and it can be the recommended randomization method for clinical trials of three treatments with equal proportion.

Objective:To investigate the correlation between the enhancement characteristics in arterial phase and the prognosis of patients with massive intrahepatic cholangiocarcinoma (IMCC).Methods:The imaging and clinical data of 92 patients with IMCC who were pathologically confirmed and underwent enhanced MRI or CT in Central Hospital of Lishui from June 2004 to February 2019 were retrospectively analyzed. According to the enhancement pattern of the primary lesion in the arterial phase, patients were divided into rich arterial blood supply group （17 cases） and deficient arterial blood supply group （75 cases）. The clinical data and imaging features of these patients were studied. Differences between measurement data and count data between the two groups were compared using t test and Fisher test. Kaplan-Meier analysis and log-rank test were used to analyze overall survival. The Cox regression multivariate analysis was used to study the relationship between the variables and the risk of death. Result:The enlargement of lymph nodes, long diameter of the primary lesion, CA19-9, treatment and HPD around the primary lesion in arterial phase were statistically different in the two groups ( P<0.05), others were no statistical difference. CA19-9>200 U/ml, lymph node enlargement, HPD around the primary lesion in arterial phase and deficient arterial blood supply were independent factors for the prediction of prognosis in IMCC patients with surgery ( P values were 0.008, 0.002, 0.049 and 0.005, respectively). Lymph node enlargement and deficient arterial blood supply were independent risk factors for the prediction of prognosis in IMCC patients with surgery ( P values are 0.049 and 0.045, respectively). Conclusion:The blood supply characteristics of arterial phase are independent factors for the prognosis of patients with IMCC.

Objective To explore the correlation between the cardiovascular ultrasound results and pathological findings of heterotaxia syndrome ,analyze the cause of the difference ,and improve the prenatal cardiovascular diagnostic accuracy of heterotaxia syndrome by ultrasound . Methods According to fetal autopsy or vessel casting , the ultrasound results of 37 fetuses with heterotaxia syndrome were retrospectively analyzed . The cardiovascular misinterpretations of ultrasound were summarized in the venous-atrialsegment,atrial-ventricular segment andventricular-arterial segment.Results ①Thirty fetuses underwent autopsy and seven fetuses vessel casting were included ,there were 10 fetuses with left atrial isomerism and 27 fetuses with right atrial isomerism .Only one left isomerism was misdiagnosed and the diagnostic accuracy of atrial isomerism by ultrasound was 97 .3％ . ② Thirty-seven fetuses with heterotaxia syndrome included 206 cardiovascular malformations . Twenty-seven misinterpretations of ultrasound were found and the rate of cardiovascular misinterpretations was 13 .1％ . ③ The cardiovascular misinterpretations of left atrial isomerism involved 1 secondary atrial septal defect in the atrial-ventricular segment and 1 aortic atresia ,2 anomalous position of arterial duct in the ventricular-arterial segment . ④The cardiovascular misinterpretations of right atrial isomerism in the venous-atrial segment included 4 common pulmonary vein atresia , 3 anomalous hepatic venous connection , 3 total abnormal pulmonary venous drainage , 2 persistent left superior vena cava and 2 abnormal inferior cava venous drainage . The cardiovascular misinterpretations of right atrial isomerism in the atrial-ventricular segment contained 2 secondary atrial septal defect ,1 unroofed coronary sinus and 1 partial atrioventricular septal defect . The cardiovascular misinterpretations of right atrial isomerism in the ventricular-arterial segment involved 3 double-outlet right ventricle ,1 truncus arteriosus and 1 anomalous position of arterial duct . Conclusions The correspondence between ultrasound results and pathological findings about cardiovascular malformations of heterotaxia syndrome is high . The most common cardiovascular misinterpretations of left atrial isomerism involve the ventricular-arterial segment . And the most common cardiovascular misinterpretations of right atrial isomerism involve the venous-atrial segment .

Objective To analyze the echocardiographic findings , associated anomalies and chromosomal characteristics in fetuses with pulmonary atresia with ventricular septal defect ( PA/VSD ) . Methods T he echocardiographic data and follow‐up materials were retrospectively reviewed in 30 256 fetuses from December 2012 to M arch 2018 in the consultation center of fetal heart disease in maternal‐fetal medicine in Anzhen hospital . Of all the fetuses ,59 cases ( 0 .19% ) had PA/VSD . T he echocardiographic findings ,associated anomalies and chromosomal characteristics were retrospectively analyzed in all the 59 fetuses with PA/VSD . Based on w hether the presence of the native pulmonary arteries and the major aortopulmonary collateral arteries ( M APCAs) or not ,the PA‐VSD was classified into type A ,type B ,and type C . Results A large ventricular defect was demonstrated in five‐chamber view with 61 .7% of the mean ratio of the aortic overriding . O ther fetal echocardiographic features of all the 59 fetuses with PA/VSD included :the right aortic arch ( n ＝19 ) ,reversal flow in the ductus arteriosus ( n ＝40 ) ,M APCAs ( n ＝24) . T he classification of the PA/VSD included :type A ( n ＝35) ,type B ( n ＝5) and type C ( n ＝19) . Associated anomalies :persistent left superior vena cava ( n ＝ 13 ) ,anomalous pulmonary vein connection ( n＝5 ) ,complete atrioventricular septal defect ( n ＝ 5 ) ; single umbilical artery ( n ＝ 3 ) ,right atrial isomerism ( n ＝3) . Of all the 30 cases performed chromosomal test ,3 cases had aneuploidy and 7 cases had microdeletion of chromosome . Conclusions The fetal echocardiographic findings of the PA/VSD are characteristic . For prenatal diagnosis of PA/VSD ,the type of PA/VSD should be defined and chromosomal test should be performed ,w hich can be helpful for prenatal consulting .

Objective To study the clinical manifestations,imaging features,treatment and outcome of umbilical venous catheters/peripherally inserted central venous catheters (UVC/PICC) associated pericardial effusion (PCE) and cardiac tamponade (CT) in neonates.Method Clinical data of cases with UVC/PICC associated PCE/CT that were found in neonatal intensive care unit of five hospitals in Shanxi province from January 2017 to December 2018 were extracted and retrospectively reviewed.Result In total,there were 632 cases received UVC/PICC insertions in 5 hospitals.7 cases of which were identified as PCE (1.0％) on echocardiography during the period of catheter indwelling,among which 5 cases suffered from PCE complicated CT (0.7％).7 cases with PCE included 6 premature infants and 1 full term infants with mean gestational age (32.6 ± 3.8) weeks and mean birth weight 1 550 (1 200,3 960) g.The mean age of PICC/UVC insertion was 14 h (1 ～ 19 h),the mean age of PCE/CT developed was 69 h (13 ～104 h) after insertion.The most common presentations included apnea/respiratory distress (7 cases),cyanosis/desaturation (7 cases),tachycardia (3 cases),bradycardia/asystole (5 cases).UVC/PICC tip ectopic was found in 6 cases after the symptoms of PCE emerged (4 cases at T6-T7 and 1 case at T12 vertebra level on chest X-ray,and 1 case at right atrium on echocardiogram).UVC/PICC tip was normal in 1 case (tip at T7-T8 vertebra level on chest X-ray).5 cases recovered after removal of catheter,pericardiocentesis and drainage of effusion;1 case recovered after removal of UVC without pericardiocentesis;1 case underwent pericardiocentesis,continued infusion for 5 h after extubated the UVC into inferior vena cava,then deceased.Conclusion UVC/PICC tip ectopic is the main cause of UVC/PICC associated PCE/CT.Immediate bedside echocardiography should be performed to any patient with UVC/PICC indwelling,who develops sudden unexplained apnea/respiratory distress,cyanosis,tachycardia/bradycardia/asystole.Timely removal of catheter and pericardiocentesis drainage may be life-saving.