Morita therapy has been bom for more than 100 years.Inpatient Morita therapy is highly oper-able and easy to master.It can improve many refractory neuroses through four-stage treatment.But more neuroses are treated in outpatient clinics,and Morita therapy cannot be used in hospitalized patients.Therefore,the formula-tion of expert opinions on outpatient operations is particularly important.This paper is based on domestic and for-eign references,and after many discussions by domestic Morita therapy experts,and then drew up the first version of the expert opinions on operation of outpatient Morita therapy.Meanwhile the operation rule of Morita therapy in three stages of outpatient treatment was formulated:in the etiological analysis stage,under the theoretical guidance of Morita therapy,analyze the pathogenic factors,to improve treatment compliance and reduce resistance;during the operating stage,guide patients to engage in constructive and meaningful actions,realizing the achievement of letting nature take its course principle;in the cultivating character and enriching life stage,pay attention to positive infor-mation,expanding the scope and content of actions,improving the ability to adapt to complex life,and preventing recurrence caused by insufficient abilities.It will lay a foundation for the promotion of Morita therapy in domestic outpatient clinics,so that more patients with neurosis and other psychological diseases could receive characteristic Morita therapy treatment in outpatient clinics.

Objective:To investigate the effects of astragaloside IV (AS-IV) on acute myocardial injury in rats with high-level spinal cord injury (SCI).Methods:Twenty-four healthy male SD rats, aged 8-10 weeks with a body weight of 250-300 g, were randomly divided into 4 groups using a random number table method: sham operation group, high-level SCI group (SCI group), high-level SCI+AS-IV group (SCI+AS-IV group) and high-level SCI+AS-IV+silent information regulator 1 (SIRT1) inhibitor EX527 group (SCI+AS-IV+EX527 group), with 6 rats in each group. The SCI model was established using the modified Allen method and the sham operation group underwent the spinal cord exposure only. In the SCI+AS-IV group, 40 mg/kg of AS-IV was injected intraperitoneally immediately after injury. SCI+AS-IV+EX527 group received an intraperitoneal injection of 5 mg/kg EX527 at one hour before injury and another injection of 40 mg/kg AS-IV in the same way immediately after injury. The sham operation group and the SCI group received an equal volume of saline via intraperitoneal injection. Immediately after awakening from injury, the hind limb motor function of the rats in each group was observed, recorded and then evaluated using the BBB method. At 24 hours after injury, the ultrastructure of the cardiomyocytes was examined under a transmission electron microscope; the levels of serum cardiac troponin I (cTnI), myocardial tissue inflammatory factors interleukin (IL)-18 and IL-1β were quantified by the ELISA method; the level of reactive oxygen species (ROS) of the myocardial tissue was assessed utilizing the dihydroethidium (DHE) assay; biochemical analyses were employed to determine the superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentrations; mRNA and protein expression levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (caspase-1), gasdermin D (GSDMD), SIRT1 and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) were examined using RT-PCR and Western blot; cardiomyocyte pyroptosis rate was evaluated by caspase-1 and TUNEL double-labeled fluorescence staining.Results:Immediately after awakening from injury, the sham operation group exhibited normal hind limb activity, with BBB scores of 21(21, 21)points, while the remaining groups displayed flaccid paralysis in both hind limbs, accompanied by the cessation of spontaneous excretion, with BBB scores of 0(0, 0)points. At 24 hours after injury, transmission electron microscopy did not reveal any significant abnormalities in the ultrastructure of the myocardiomyocytes in the sham operation group, while changes of varying degrees were observed in the SCI group. The ELISA results indicated that at 24 hours after injury, the serum cTnI level in the SCI group was (1 435.3±148.1)pg/ml, higher than (619.6±95.4)pg/ml in the sham operation group ( P<0.01); the cTnI level was (1 154.0±80.0)pg/ml in the SCI+AS-IV group, lower than that in the SCI group ( P<0.01); the cTnI level was (1 321.8±50.2)pg/ml in the SCI+AS-IV+EX527 group, higher than that in the SCI+AS-IV group ( P<0.05). The levels of IL-18 and IL-1β in the myocardial tissue in the SCI group were (493.0±145.0)pg/ml and (936.7±93.2)pg/ml, higher than (131.1±62.5)pg/ml and (281.7±83.6)pg/ml in the sham operation group ( P<0.01); the levels of IL-18 and IL-1β in the SCI+AS-IV group were (182.4±45.6)pg/ml and (573.4±99.5)pg/ml, lower than those in the SCI group ( P<0.01); the levels of IL-18 and IL-1β in the SCI+AS-IV+EX527 group were (337.4±72.0)pg/ml and (742.6±82.7)pg/ml, higher than those in the SCI+AS-IV group ( P<0.05), yet lower than those in the SCI group ( P<0.01). At 24 hours after injury, DHE and biochemical assays showed that the levels of ROS and MDA in the myocardial tissue in the SCI group were (65±6)% and (1.97±0.27)nmol/mg, higher than (19±10)% and (1.03±0.16)nmol/mg in the sham operation group ( P<0.01); the ROS and MDA levels in the SCI+AS-IV group were (37±10)% and (1.39±0.11)nmol/mg, lower than those in the SCI group ( P<0.01); the ROS and MDA levels in the SCI+AS-IV+EX527 group were (52±7)% and (1.70±0.14)nmol/mg, higher than those in the SCI+AS-IV group ( P<0.05). The SOD level in the myocardial tissue of the SCI group was (658.48±77.56)U/mg, lower than (1 059.55±71.91)U/mg in the sham operation group ( P<0.01); the SOD level in the SCI+AS-IV group was (901.74±32.30)U/mg, higher than that in the SCI group ( P<0.01); the SOD level in the myocardial tissue in the SCI+AS-IV+EX527 group was (799.86±26.70)U/mg, lower than that in the SCI+AS-IV group ( P<0.05). At 24 hours after injury, RT-PCR showed that the mRNA expression levels of NLRP3, caspase-1 and GSDMD in the myocardial tissue of the SCI group were 2.07±0.25, 2.46±0.28 and 1.82±0.12 respectively, which were higher than 1.10±0.13, 0.95±0.17 and 1.03±0.08 in the sham operation group ( P<0.01); the mRNA expression levels of NLRP3, caspase-1 and GSDMD in the SCI+AS-IV group were 1.47±0.24, 1.51±0.16 and 1.42±0.13 respectively, which were lower than those in the SCI group ( P<0.01); the mRNA expression levels of NLRP3, caspase-1 and GSDMD in the SCI+AS-IV+EX527 group were 1.93±0.28, 1.97±0.31 and 1.65±0.16 respectively, which were higher than those in the SCI+AS-IV group, yet lower than those in the SCI group ( P<0.05). The mRNA expression levels of SIRT1 and PGC-1α in the myocardial tissue in the SCI group were 0.41±0.09 and 0.56±0.07, lower than 1.20±0.14 and 1.29±0.20 in the sham operation group ( P<0.01); the mRNA expression levels of SIRT1 and PGC-1α in the myocardial tissue in the SCI+AS-IV group were 0.78±0.08 and 1.01±0.19, higher than those of the SCI group ( P<0.01); the mRNA expression levels of SIRT1 and PGC-1α in the myocardial tissue of the SCI+AS-IV+EX527 group were 0.53±0.12 and 0.72±0.22, lower than those of the SCI+AS-IV group ( P<0.05). At 24 hours after injury, the western blot analysis showed that the protein expression levels of NLRP3, caspase-1 and GSDMD in the myocardial tissue in the SCI group were 1.00±0.20, 0.60±0.19 and 0.77±0.15 respectively, which were higher than 0.27±0.09, 0.18±0.10 and 0.28±0.08 in the sham operation group ( P<0.01); the protein expression levels of NLRP3, caspase-1 and GSDMD in the SCI+AS-IV group were 0.59±0.10, 0.25±0.11 and 0.33±0.11 respectively, lower than those in the SCI group ( P<0.01); the protein expression levels of NLRP3, caspase-1 and GSDMD in the myocardial tissue in the SCI+AS-IV+EX527 group were 0.85±0.15, 0.54±0.12 and 0.55±0.13 respectively, higher than those in the SCI+AS-IV group ( P<0.05). The protein expression levels of SIRT1 and PGC-1α in the myocardial tissue in the SCI group were 0.44±0.16 and 0.28±0.10, lower than 0.93±0.22 and 0.75±0.16 in the sham operation group ( P<0.01); the protein expression levels of SIRT1 and PGC-1α in the myocardial tissue in the SCI+AS-IV group were 0.78±0.19 and 0.55±0.12, higher than those in the SCI group ( P<0.01); the protein expression levels of SIRT1 and PGC-1α in the myocardial tissue in the SCI+AS-IV+EX527 group were 0.46±0.16 and 0.35±0.07, lower than those in the SCI+AS-IV group ( P<0.05). At 24 hours after injury, caspase-1 and TUNEL double-labeled fluorescence staining showed that the cardiomyocyte pyroptosis rate in the SCI group was (34.5±6.7)%, higher than (5.3±2.9)% in the sham operation group ( P<0.01); the cardiomyocyte pyroptosis rate in the SCI+AS-IV group was (13.4±3.0)%, lower than that in the SCI group ( P<0.01); the cardiomyocyte pyroptosis rate in the SCI+AS-IV+EX527 group was (22.5±5.9)%, higher than that in the SCI+AS-IV group ( P<0.01), yet lower than that in the SCI group ( P<0.01). Conclusions:AS-IV can significantly reduce acute myocardial injury in rats with high-level SCI. Its mechanism may involve activating the myocardial SIRT1/PGC-1α signaling pathway, protecting the mitochondria, enhancing the ability to resist oxidative stress, and effectively inhibiting the NLRP3 inflammasome-mediated pyroptosis pathway.

ObjectiveTo observe the effect of augmented reality training based on enriched environment on walking dysfunction after stroke. MethodsFrom January, 2021 to June, 2022, 36 stroke patients in the Affiliated Suzhou Hospital of Nanjing University Medical School were randomly divided into control group (n = 18) and experimental group (n = 18). Both groups received conventional rehabilitation treatment. The control group was supplemented with conventional walking training, and the experimental group was supplemented with augmented reality training based on enriched environment, for four weeks. They were assessed with Berg Balance Scale (BBS), Timed Up and Go Test (TUGT), 10-meter walk test (10MWT) and Barthel Index (BI) before and after treatment, and the gait parameter was compared. ResultsNo adverse event occurred during treatment. After treatment, the BBS score, TUGT time, 10MWT speed, BI, gait speed, gait frequency and the proportion of single-leg support on the affected side significantly improved in both groups (|t| > 5.161, P < 0.001). All the above indexes were better in the experimental group than in the control group (|t| > 2.106, P < 0.05), except for BI (t = 1.099, P = 0.282). ConclusionAugmented reality training based on enriched environment could improve the walking function of paitents after stroke, which is better than conventional walking training.

Objective:To explore any effect of aerobic exercise on cardiac energy metabolism and mitochondrial respiration after myocardial infarction.Methods:Forty-five Sprague-Dawley rats were randomly divided into a sham operation group, a heart failure control group and a heart failure exercise group. Myocardial infarction was induced in the heart failure groups using coronary artery ligation. Four weeks after the successful modeling, the heart failure exercise group underwent 8 weeks of aerobic treadmill exercise. The cardiac function and exercise ability of all of the rats were then observed using echocardiography and the incremental treadmill exercise test. Myocardial creatine phosphate (PCr) and adenosine triphosphate (ATP) were measured by magnetic resonance spectroscopy, and the respiratory function of the myocardial mitochondria was evaluated by using cell respirometry.Results:Compared with the sham operation group, the average PCr content, PCr/ATP ratio, oxygen consumption of mitochondrial respiratory chain complexes I and II, left ventricular shortening fraction (FS) and ejection fraction (EF), maximum running speed, exhaustion distance and exhaustion time in the incremental treadmill exercise test were all significantly worse in the heart failure control group. Moreover, the average ATP content, complex I oxygen consumption, left ventricular FS and EF, and the maximum running speed, exhaustion distance and exhaustion time in incremental treadmill exercise of the heart failure exercise group were all superior to those of the heart failure control group.However, no significant differences were observed in the average PCr/ATP ratio between the heart failure exercise and control groups.Conclusions:Regular aerobic exercise can improve cardiac performance after chronic heart failure, at least in rats. The mechanism may be related to increased levels of myocardial ATP and better mitochondrial complex I functioning. The PCr/ATP ratio may not be a suitable biomarker for evaluating the benefits of exercise for the heart.

Objective:To investigate the changes of autonomic nervous active substances in myocardium of rats with acute high-level spinal cord injury.Methods:Twenty-four clean-level healthy adult male SD rats weighting 250-300 g for 8-10 weeks old were divided into control group ( n=6) and spinal cord injury group ( n=18) according to the random number table. The spinal cord injury group was subdivided at 4, 12 and 24 hours, with 6 rats at each time point. The high-level spinal cord injury model was established by the modified Allen′s weight-drop method, and the spinal cord was only exposed in control group. The postoperative performance and BBB score for limb movement were observed in each group. The myocardium of each group was resected and used to observe ultrastructure of myocardial cells under transmission electron microscope and detect protein and mRNA levels of tyrosine hydroxylase (TH), noradrenaline transporter (NET), acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) by Western blot and RT-PCR analysis. Results:Rats of control group showed normal limb motion after operation without significant change from the preoperation level, and mean BBB score was 21 points. Rats of spinal cord injury group showed significantly reduced activities and feeding, with flaccid paralysis of both lower limbs as well as no spontaneous excretion, and showed BBB score of 0 point at 4 hours and 12 hours after injury, which was increased slightly at 24 hours after injury, with the highest score for 1 point. The ultrastructure of myocardial cells showed no obvious abnormalities in control group, while different degrees of changes in spinal cord injury group. Compared with control group, Western blot analysis showed that protein levels of TH and NET were decreased, while AChE and ChAT were increased in spinal cord injury group ( P<0.05 or 0.01). Compared with control group, RT-PCR analysis showed that mRNA levels of TH and NET were decreased, while AChE and ChAT were increased in spinal cord injury group ( P<0.05 or 0.01). mRNA levels of TH and NET in spinal cord injury group at 24 hours after injury were significantly different from those at 4 hours and 12 hours after injury (all P<0.05). mRNA levels of ChAT in spinal cord injury group were statistically significant at 12 hours and 24 hours after injury from those at 4 hours after injury, with significant difference at 12 hours and 24 hours after injury (all P<0.05). Conclusion:Sympathetic nerve active substances TH and NET are down-regulated but vagal nerve active substances AChE and ChAT up-regulated in myocardium of rats with acute high-level spinal cord injury, which may be related to the relative excitation of the parasympathetic nerve blocking the sympathetic innervation of the higher center to the heart following high-level spinal cord injury.

Objective:To explore the effect of shoulder subluxation on the peripheral nerves in the hemiplegic upper limbs of stroke survivors.Methods:Twenty stroke survivors with shoulder subluxation were enrolled. Conduction in their suprascapular, axillary, musculocutaneous, radial, median and ulnar nerves was monitored and needle electromyography was used to monitor activity in the supraspinatus, deltoid, biceps brachii, extensor digitorum, abductor pollicis brevis and abductor digiti minimi muscles of their affected upper limbs at rest. Upper limb and hand function were assessed using the Brunnstrom scale. The rate of change in the amplitude of the compound muscle action potentials (CMAPs) was correlated with the patient′s disease duration, age, and upper limb and hand Brunnstrom stages.Results:Compared with the healthy side, a significant decrease was observed in the CMAP amplitudes of the suprascapular, axillary, musculocutaneous, radial, median and ulnar nerves of the hemiplegic arm, and the latency of the suprascapular and axillary nerves was significantly prolonged. There was no inter-arm difference in the conduction velocity of the musculocutaneous, radial, median and ulnar nerves. The rates of change in the CMAP amplitudes of the suprascapular, axillary and musculocutaneous nerves were significantly higher than those of the radial, median and ulnar nerves. The sensory nerve action potential (SNAP) amplitudes of the median, ulnar and radial nerves on the hemiplegic side were significantly lower than on the healthy side, but there was no significant difference in the sensory conduction velocity between the two sides. On the hemiplegic side, the median nerve had the highest rate of change rate in the SNAP amplitude, followed by the radial and ulnar nerves, but there was no significant difference among them. Nor was there any significant difference in the rate of change in sensory nerve conduction velocity. The muscles of the affected upper limbs had higher potentials in the proximal than that in the distal nerves after shoulder subluxation. The rate of change in the CMAPs was not significantly correlated with a patient′s disease duration, age, or upper limb or hand Brunnstrom stage on the hemiplegic side.Conclusions:Shoulder subluxation after a stroke can cause greater damage to the peripheral nerves in the shoulder and upper arm than to those in the forearm and hand, possibly affecting the recovery of upper limb function.

Objective:To investigate the incidence of high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangement in Chinese diffuse large B-cell lymphoma (DLBCL) .Methods:From January 2013 to August 2020, 922 DLBCL cases were collected. C-MYC and BCL2 protein expression levels were analyzed by immunohistochemistry staining. Fluorescence in situ hybridization was used to detect the structural abnormalities of MYC, BCL2, and BCL6, including gene breaks and copy number changes.Results:MYC and BCL2 and/or BCL6 gene breaks were found in 29 out of 922 DLBCL cases (3.15%) , including 25 cases of double-hit lymphoma (DHL; 14 cases involving MYC and BCL2 rearrangements and 11 cases involving MYC and BCL6 rearrangements) and four cases involving MYC, BCL2, and BCL6 rearrangements, referring to triple-hit lymphoma. According to the threshold of C-MYC ≥40% and BCL2 ≥50%, 541 cases (58.68%) overexpressed C-MYC and BCL2 proteins, including 22 DHL cases. Moreover, according to the threshold of C-MYC ≥70% and BCL2 ≥50%, 52 cases (5.64%) overexpressed C-MYC and BCL2 proteins, including nine DHL cases. The P53 protein expression was detected by immunohistochemistry staining. The mutant P53 expression pattern was shown in 101 out of 709 cases (14.25%) , whereas 13 cases (1.83%) were negative, likely indicating P53 gene fragment deletion.Conclusion:The incidence of high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements was low in DLBCLs, and no significant correlation between gene abnormality and protein overexpression was shown. The correct diagnosis of DHL depends on molecular genetic detection.

Objective: To understand the epidemiological characteristics of outbreaks and analyze the genetic characteristics of the whole genome of influenza H3N2 virus among avian-human-swine, and to elaborate the source of influenza virus. Methods: The epidemic information was collected using the case investigation, the pharyngeal swab samples from influenza-like-illness cases were detected by real-time PCR and virus isolation. The phylogeny and molecular features of whole-genome were analyzed with EditSeq and MEGA 5.05 software. Results: The prevalence rate of this outbreak was 34.88%, 15 samples of throat swabs were collected, the positive rate of nucleic acid detection was 73.33%, 5 strains of seasonal influenza A (H3N2) influenza viruses were isolated. The phylogenetic analysis showed the eight gene segments of the isolated influenza viruses belonged to the same cluster with 2015-2016 influenza vaccine strain A/Switzerland/9715293/2013(H3N2), and no recombination was found. Compared with vaccine strain, 14 variant amino acids of protein of HA were identified, and 8 of them were located in antigenic sites. All strains were sensitive to neuraminidase inhibitors while they showed resistance to blockers of M2 ion channel. The glycosylation sites analysis showed that two new glycosylation sites NRT151-153 and NAT245-247were added. Conclusions The outbreak was caused by seasonal influenza A (H3N2) virus which had an antigenic drift and no genetic avian-human-swine recombination was found.

Objective To explore the correlations of vascular endothelial growth factor (VEGF) and paraoxonase 1 (PON1) gene polymorphism with diabetic retinopathy. Methods A total of 168 patients with diabetes mellitus(DM) in the Second People's Hospital of Xining City were selected from January 2013 to December 2015, and were divided into the DM group (48 cases), proliferative diabetic retinopathy (PDR) group (66 cases) and non-proliferative diabetic retinopathy (NPDR) group (54 cases). A total of 104 healthy subjects were selected as the control group (NC group). The VEGF and PON1 gene polymorphisms were detected by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and levels of superoxide dismutase (SOD) and malondialdehyde (MDA) of all subjects were detected by using UV spectrophotometer. Results Compared with the NC group, the systolic blood pressures and glycated hemoglobin levels in the DM group, NPDR group and PDR group were significantly increased,there were statistically significant differences (P<0.05). No statistically significant difference was found in diastolic blood pressure and levels of cholesterol,triglyceride, high density lipoprotein among these groups (P>0.05). There was statistically significant difference in systolic blood pressure between PDR group and NPDR group (P< 0. 05), while no statistically significant difference was found in diastolic blood pressure and levels of cholesterol, triglyceride,high density lipoprotein and glycated hemoglobin between the two groups (P>0.05). The incidence rate of DR in the patients with CC,CT and TT genotype was 71.05%, 56.27 % and 38.64 %, respectively. The incidence rate of DR in the patients with CCgenotype was significantly higher than that in the patients with CT or TT genotype (P< 0.05). No statistically significant difference was found in SOD and MDA levels of healthy subjects with different PON1 192 genotypes (P>0.05). The SOD level of DR patients with QQ genotype was lower than that of patients with QR or RR genotype,and the MDA level of DR patients with QQ genotype was higher than that of patients with QR or RR genotype,the difference was statistically significant (P<0.05). Conclusion The expression of VEGF and PON1 genes could affect the development and progression of DR. PON1 gene might control the progression of DR by affecting the expression of oxidative kinase in DR patients.

OBJECTIVETo investigate the expression of high mobility group box 1 (HMGB1) in gingival tissues of chronic periodontitis.

METHODSHuman peripheral blood mononuclear cells(PBMC) were stimulated with 1 microg x mL(-1) lipopolysaccharide (LPS) for 24 h or 48 h. Expression and release of HMGB1 were checked by immunofluorescence and enzyme-linked immunosorbent assay (ELISA), respectively. PBMC were stimulated with 100 ng x mL(-1) HMGB1 or 50 ng x mL(-1) tumor necrosis factor-alpha (TNF-alpha), the expressions of TNF-alpha and HMGB1 in the supernatant were studied by ELISA. Gingival tissues and gingival crevicular fluids (GCF) were collected from patients and healthy people. Expression of HMGB1 in gingival tissues and GCF was studied using immunofluorescence and ELISA, respectively.

RESULTSHMGB1 was translocated from nucleus to cytosol in PBMC after LPS stimulation for 24 h. The content of HMGB1 in the supernatant from stimulated cells was significantly higher than that from unstimulated cells after 48 h (P < 0.01). HMGB1 was released by PBMC in response to TNF-alpha stimulation, it also stimulated PBMC to release TNF-alpha (P < 0.01). Translocation of HMGB1 from nucleus to cytosol was also found in infiltrated cells in gingival tissues from patients, and HMGB1 in GCF from patients was significantly higher than that from healthy people P < 0.01).

CONCLUSIONThe results suggest that HMGB1 may play an important role in the pathological progress of chronic periodontitis.