Objective To observe the effects of electroacupuncture at"Ciliao","Zhongji","Sanyinjiao"and"Dazhui"on urodynamics and expression of ERK/CREB/Bcl-2 pathway in spinal cord tissue of neurogenic bladder rats after suprasacral spinal cord injury.Methods Sixty female SD rats randomly selected 24 and divided into blank group and sham-operation group(12 rats in each group),the remaining 36 rats were subjected to surgical modeling.After modeling,rats were randomly divided into the model group and the electroacupuncture group,with 12 rats in each group.The electroacupuncture group received unilateral electroacupuncture stimulation at acupoints"Ciliao","Zhongji","Sanyinjiao",and"Dazhui"for 30 minutes each time,once a day,for 7 consecutive days.After administration,urodynamic testing was performed,HE staining was used to observe the morphology of bladder detrusor tissue,TUNEL method was used to detected apoptosis in spinal cord tissue,Western blot was used to detected expressions of p-ERK1/2,p-CREB,p-p90Rsk,CRE,Bcl-2,and Bax proteins in spinal cord tissue.Results Compared with the sham-operation group,the basal pressure,maximum pressure,and leakage point pressure of the bladder in the model group increased significantly(P<0.01),while the maximum capacity and compliance of the bladder decreased significantly(P<0.01);the structure of bladder smooth muscle cells was severely damaged and disorderly arranged,accompanied by a large amount of inflammatory cell infiltration;the apoptosis rate of spinal cord tissue cells significantly increased(P<0.01),and the expressions of p-ERK1/2,p-p90Rsk,p-CREB,CRE,and Bcl-2 proteins in spinal cord tissue were significantly decreased,while the expression of Bax protein significantly increased(P<0.01).Compared with the model group,the basal pressure,maximum pressure,and leakage point pressure of the bladder in the electroacupuncture group decreased significantly(P<0.05),while the maximum capacity and compliance of the bladder increased significantly(P<0.05,P<0.01);the integrity of bladder smooth muscle cells was enhanced,the degree of cell edema was reduced,and inflammatory cell infiltration was reduced;the apoptosis rate of spinal cord tissue cells was significantly reduced(P<0.05),and the expressions of p-ERK1/2,p-p90Rsk,p-CREB,CRE,and Bcl-2 proteins in spinal cord tissue significantly increased,while the expression of Bax protein was significantly decreased(P<0.05,P<0.01).Conclusion Electroacupuncture can promote the repair of bladder detrusor tissue in rats with neurogenic bladder model after suprasacral spinal cord injury,increase the maximum capacity and compliance of the bladder,alleviate the high pressure state in the bladder,and its mechanism is related to activating the ERK/CREB/Bcl-2 pathway,reducing secondary apoptosis of damaged neurons,effectively improving bladder innervation,and protecting bladder function.

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 clarify the preparation methods of four rat models of liver ischemia/reperfusion injury (IRI) and to determine a liver IRI animal model that is consistent with clinical conditions, has stable pathological and physiological injury, and is easy to operate. METHODS: A total of 160 male Sprague-Dawley (SD) rats were randomly divided into four groups using an interval grouping method: 70% IRI (group A), 100% IRI (group B), 70% IRI with 30% hepatectomy (group C), and 100% IRI with 30% hepatectomy (group D), with 40 rats in each group. Each model was further divided into sham operation group (S group) and ischemia groups of 30, 60, and 90 minutes, with 10 rats in each group. After surgery, the survival status and awakening time of the rats were observed, and the liver lobectomy weight, bleeding volume, and hemostasis time of groups C and D were recorded. Blood samples were collected by cardiac puncture after 6 hours of reperfusion for determination the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), blood urea nitrogen (BUN), serum creatinine (SCr), and γ-glutamyl transpeptidase (γ-GT) in the serum to assess liver and kidney function. Hematoxylin-eosin (HE) staining and immunohistochemical staining of macrophages were performed to analyze the liver tissue structure damage from a pathological perspective. RESULTS: Rats in group A exhibited earlier awakening and acceptable mental status, while rats in the other groups showed delayed awakening and poor mental status. The hemostasis time in group D was approximately 1 second longer than that in group C. The mortality of rats subjected to 60 minutes of 70% hepatic ischemia was 0. Compared to the sham operation group, rats in each experimental group showed significant increases in serum levels of AST, ALT, ALP, BUN, SCr, and γ-GT, indicating impaired liver and kidney function in the rat models of liver IRI. In groups A, B, and C, the 90-minute ischemia subgroup exhibited more pronounced elevation in AST, ALT, ALP, BUN, SCr, and γ-GT levels compared to the 30-minute ischemia subgroup [AST (U/L): group A, 834.94±56.73 vs. 258.74±18.33; group B, 547.63±217.40 vs. 277.67±57.92; group C, 930.38±75.48 vs. 640.51±194.20; ALT (U/L): group A, 346.78±25.47 vs. 156.58±13.25; group B, 408.40±138.25 vs. 196.80±58.60; group C, 596.41±193.32 vs. 173.76±72.43; ALP (U/L): group A, 431.21±34.30 vs. 315.95±15.64; group B, 525.88±62.13 vs. 215.63±17.31; group C, 487.53±112.37 vs. 272.46±92.33; BUN (U/L): group A, 18.35±5.63 vs. 14.32±2.30; group B, 30.21±4.55 vs. 17.41±8.14; group C, 20.50±3.64 vs. 15.93±3.22; SCr (U/L): group A, 27.47±8.91 vs. 22.37±5.66; group B, 43.60±15.57 vs. 36.80±7.95; group C, 63.81±20.24 vs. 42.47±7.03; γ-GT (U/L): group A, 15.64±3.57 vs. 6.82±1.48; group B, 9.28±1.91 vs. 5.62±1.21; group C, 10.98±3.18 vs. 5.67±1.10; all P < 0.05]. The 100% IRI 90-minute group and 100% IRI 90-minute group with 30% hepatectomy exhibited more pronounced increases in the above-mentioned indicators compared to the corresponding 70% IRI control group, indicating increased liver and kidney damage in rats subjected to combined blood flow occlusion and hepatectomy. HE staining showed clear liver tissue structure with intact and orderly arranged cells in the sham operation group, while the experimental groups exhibited cell structure damage, including cell rupture or collapse, cell swelling, nuclear pyknosis, deep cytoplasm staining, cell shedding, and necrosis. The interstitium showed infiltration of inflammatory cells. Immunohistochemical staining revealed a higher number of macrophages in the experimental groups compared to the sham operation group. CONCLUSIONS Four models of liver IRI in rat were successfully established. As the duration and severity of hepatic ischemia increased, liver cell ischemia worsened, leading to increased hepatocellular necrosis and exhibiting characteristic features of liver IRI. These models can effectively simulate liver IRI following liver trauma, with the group subjected to 100% ischemia and 30% hepatectomy showing the most severe liver injury. The designed models are reasonable, easy to perform, and exhibit good reproducibility. They can be used for investigating the mechanisms, therapeutic efficacy, and diagnostic methods related to clinical liver IRI.
Rats ; Male ; Animals ; Reproducibility of Results ; Rats, Sprague-Dawley ; Liver ; Reperfusion Injury/drug therapy* ; Ischemia ; Disease Models, Animal ; Necrosis

OBJECTIVE: To summarize the genetic characteristics of 671 Chinese pedigrees affected with Duchenne/Becker muscular dystrophy (DMD/BMD). METHODS: Clinical data of the pedigrees were collected. Multiplex PCR, multiple ligation dependent probe amplification (MLPA), next generation sequencing (NGS), Sanger sequencing and long read sequencing were used to detect the variant of DMD gene in the probands and their mothers, and prenatal diagnosis was provided for high risk pregnant women. RESULTS: Among 178 pedigrees analyzed by multiplex PCR, 44 variants of the DMD gene were detected, with the genetic diagnosis attained in 110 pedigrees. Among 493 pedigrees analyzed by MLPA in combination with NGS or Sanger sequencing, 294 pathogenic/possible pathogenic variants were identified, among which 45 were unreported previously, and the genetic diagnosis attained in 484 pedigrees. Structural variants of the DMD gene were identified in two pedigrees by long-read sequencing. Among 444 probands, 341 have inherited the DMD gene variant from their mothers (76.8%). Among 390 women with a high-risk, 339 have opted to have natural pregnancy and 51 chose preimplantation genetic testing for monogenetic disease (PGT-M). The detection rate of neonatal patients and carriers following natural pregnancy was significantly higher than that for PGT-M. CONCLUSION Combined application of MLPA, NGS, Sanger sequencing and long-read sequencing is an effective strategy to detect DMD/BMD. PGT-M can effectively reduce the risk of fetuses. Above finding has expanded the spectrum of DMD gene variants and provided a basis for reproductive intervention for pregnancies with a high risk for DMD/BMD.
China ; Dystrophin/genetics* ; Exons ; Female ; Genetic Testing ; Humans ; Infant, Newborn ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne/genetics* ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis

Objective Colon-targeting capsules based on gastric pellets and enteric pellets were prepared from Baizhu Huanglian prescription. The formulation composition and preparation process were optimized and the in-vitro release characteristics were investigated. Methods Optimum formulation composition and process parameters of Baizhu Huanglian pellets were screened out by single factor experiment and orthogonal design. The pellets core were prepared by extrusion-spheronization technique and coated in the fluid bed using bottom spray coating technique. To investigate the effect of coating level of the isolation layer, the proportion of polymer, the amount of plasticizer and weight gain of enteric coating on the release behavior of the enteric pellets. The pellets release behavior was fitted by model as well. Results The prescription of gastric pellets was drug loading 50%, PVPP 5%, MCC to lactose 1∶2 and wetting agent 40%. The process parameters were extrusion frequency 20 Hz, rounding speed 500 r/min and rounding time 5 min. The prescription of enteric pellets was drug loading 27%, PVPP 5%, MCC to lactose 5∶2, wetting agent 30% and adhesive 20%. The process parameters were extrusion frequency 20 Hz, rounding speed 700 r/min and rounding time 7 min. For enteric coating layer, the coating mixture of EUDRAGIT®L30D-55 to EUDRAGIT® FS30D was 1∶2. The amount of plasticizer was 10%. The increased weight of coating layer was 15%. The release time of enteric pellets in-vitro was up to 24 hours. The release behavior of the pellets conforms to the Higuchi model. Conclusion The colon targeting capsule of Baizhu Huanglian pellets were successfully prepared and showed the characteristics of sustained release and colon targeting.

Objective:To investigate the clinical features and influencing factors of renal artery involvement in patients with Takayasu's arteritis (TA).Methods:One hundred and five TA patients were included and their clinical features, laboratory tests, echocardiographic findings were collected and analyzed. The t-test, Mann-Whitney U test, Sperman's correlation and logistic regression were used for statistical analysis. Results:Of the 105 patients with Takayasu's arteritis, 21 (20%) patients were diagnosed to have renal artery involvement. They had higher frequency of hypertension [76%(16/21) vs 25%(21/84); χ2=19.291, P<0.01], syncope [19%(4/21) vs 4%(3/84); P=0.028] and headache [52%(11/21) vs 25%(21/84); χ2=5.944, P=0.019]. Patients with renal artery invol-vement had higher levels of creatinine [(67±30) mmol/L vs (50±14) mmol/L; t=2.436, P=0.023], lower levels of lymphocyte subsets CD3 +[1 299(1 261.5, 1 313.45)/μl vs 1 531 (1 330, 1 559.5)/μl; Z=-3.12, P=0.002], CD4 +[793.6(715, 804.32)/μl vs 914.88(794.3, 914.8)/μl; Z=-2.597, P=0.009], CD8 +[451.09(451.09, 489.5)/μl vs 552.39(459.75, 557.5)/μl; Z=-3.271, P=0.001] and glomerular filtration rate ( Z=-2.612, P=0.009). Comparison of echocardiographic parameters between the two groups showed that the diameters of left atrium [(35±4) mm vs (32±4) mm; t=3.065, P=0.003] and end-diastolic left ventricular [49.2(47.5, 52.5) vs 45.84(43.25, 47.75); Z=-3.795, P<0.01] were larger in renal artery involvement group. Abdominal aorta ( χ2=14.292, P<0.01), superior mesenteric artery ( P<0.01), inferior mesenteric artery ( P=0.038) and common iliac artery ( P=0.005), were more likely involved in patients with renal artery involvement; while the common carotid artery ( χ2=3.815, P=0.047) and subclavian artery ( P=0.022) were less affected. There was a positive correlation between creati-nine level ( r=0.282, P=0.004) and renal artery involvement in patients with arteritis, and the number of lymphocyte subsets (CD3, CD4, CD8) ( r=-0.306, P=0.002; r=-0.255, P=0.009; r=-0.321, P=0.001) was negatively correlated with renal artery involvement. Conclusion:The most common pattern of renal artery involvement is stenosis and occlusion. Hypertension, syncope, headache, kidney dysfunction and heart failure are more prevalent among patients with renal artery involvement. The number of lymphocyte subsets is low, and involvement of abdominal aorta is common. It should be assessed and treated as soon as it is discovered.

Objective: To introduce a combined operation for treating chronical peroneal tendon dislocation and to evalu-ate the clinical outcomes of patients. Methods: Data of 12 ankles in 12 patients (male 9, female 3) with chronical peroneal ten-don dislocation who underwent the fibular groove deepening procedure with transposition of the peroneal longus muscle from June 2006 to August 2013 were retrospectively analyzed. The mean age was 24.6±4.3 years (range, 18-34 years), and there were 3 cas-es on left side, 9 on right side. The mechanism of peroneal tendon injury consisted of sports injuries in 8 and sprains in 4. There were 8 cases of flat fibula sulcus and 4 cases of convex. All patients met the inclusion criteria of a painful snapping or popping sen-sation or palpable clicking and positive provocation maneuver and without fracture and were treated with peroneal sulcus deepen-ing and peroneal longus tendon transposition. The duration of preoperative popping and pain symptoms ranged from 12 to 23 weeks, with an average of 16.9±4.0 weeks. All cases were treated conservatively 3-4 weeks before operation and were not effec-tive. The clinical outcomes were evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot scale, visual anologue scale (VAS). Results: Twelve patients were followed up with an average period of 37.3±7.0 (range, 25-50) months. The mean VAS scale score of all patients reduced from 5.6±0.9 to 0.5±0.7 at the latest follow-up. The mean AOFAS scale score improved from 61.4±5.6 to 92.6±4.2 at the latest follow-up. The difference between preoperative and postoperative was statis-tically significant. (t=16.250,-18.475; P=0.000). According to the evaluation of symptom and function scoring system, 10 cases were excellent and 2 cases were good, with an excellent rate of 100%. The mean postoperative return-sport time was 26.42±3.06 weeks (range, 23-32 weeks). All patients healed primarily and no infection, skin necrosis and residual redislocation occurred. The osteotomy healed completely without displacement, which was confirmed by imaging examination in three months postoperation. No patients had intractable pain after surgery, and they were able to perform daily activities at 3 months and physical exercise at 6 months after surgery. No slip occurred in all patients at the latest follow-up. Conclusion The fibular groove deepening procedure with transposition of the peroneal longus muscle can effectively treat chronical peroneal tendon dislocation and obtain good medi-um-term results.

Objective:To explore the relationship between the expression of p57KIP2,cyclin D1,and cyclin E receptors in male breast can-cer(MBC)and its clinical significance.Methods:Data of MBC cases diagnosed in The First Affiliated Hospital of Wenzhou Medical Uni-versity between January 2000 and December 2016 were reviewed.Forty cases of infiltrating ductal carcinoma(IDC)and 20 cases of male ductal carcinoma in situ(DCIS)were selected,and 20 cases of gynecomastia(GYM)were selected as controls.The expression of p57KIP2,cyclin D1,and cyclin E mRNA and protein in the tissue samples obtained from IDC,DCIS,and GYM cases were measured by re-verse transcription polymerase chain reaction and immunohistochemistry,respectively.Results:The expression of p57KIP2mRNA in IDC was 0.18±0.07,which was lower than that in DCIS(0.42±0.05)and GYM(0.75±0.04).The rate of p57KIP2positivity in IDC was 25%(10/40),which was lower than that in DCIS 60%(12/20)and GYM 90%(18/20).There were significant differences in the expression of p57KIP2mRNA and protein among the three groups(P<0.05).The expression of cyclin D1 and cyclin E mRNA in IDC was 0.92±0.12 and 0.96±0.08,which was higher than that in DCIS(0.72±0.06,0.64±0.01)and GYM(0.38±0.03,0.21±0.02),respectively.Cyclin D1 and cy-clin E protein positive rates in IDC were 90%(36/40)and 88%(35/40),which were higher than those in DCIS[80%(16/20),85%(17/20)],and GYM[25%(5/20),20%(4/20)].In IDC tissues,the expression of p57KIP2,cyclin D1,and cyclin E proteins was associated with the clinical stage and histological grade(P<0.05),and the expression of p57KIP2protein was correlated with axillary node metastasis(P<0.05).There was a negative correlation between the expression of p57KIP2and cyclin D1 and between p57KIP2and cyclin E.However,the correlation between cyclin D1 and cyclin E expression was positive(P<0.05).Conclusions:p57KIP2,cyclin D1,and cyclin E may play an im-portant role in the development and progression of MBC.Combined clinicopathological detection of p57KIP2,cyclin D1,and cyclin E can aid future research on MBC.

Objective To study the mental status and its influencing factors of patients with rheumatic disease. Methods Three hundreds and eighty-one patients with rheumatic diseases were assessed with the self-rating anxiety scale, self-rating depressive scale, Eysenck Personality Questionnaire symptom checklist-90, and demographic characteristics questionnaire. The comparison among groups were tested by t tests, Spearman's correlation analysis. Results Patients with anxiety symptoms accounted for 35.96%, and pre-valence rate of depression was 48.03%. The gender (r=0.87, OR=2.34) and duration (r=0.006, OR=1.006) were the risk factors and the C4 (r=-2.63, OR=0.07) was the protective factor. In patients with systemic lupus erythematosus, depression was related to education (Z=2.228, P=0.026), disease duration (Z=-2.015,P=0.044), IgG level (t=-2.266, P=0.027), T cell subsets CD8 (Z=1.981, P=0.047) and CD3 (Z=1.967, P=0.049). Anxiety was related to education (Z=2.452, P=0.014), duration (Z=-2.054, P=0.04), complement C3 (t=2.035, P=0.045), C4 (Z=-2.204, P=0.028) and IgG level (t=-2.126, P=0.036). The value of CRP(Z=-1.984, P=0.047) and IgM (t=2.243, P=0.028) of rheumatoid arthritis patients were higher in the depression group. The depression group was different in disease duration (Z=-2.631, P=0.009), gender ( χ2=4.415, P=0.042), ESR (Z=2.17, P=0.023) and CD3 cell (Z=2.023, P=0.043) in Sj?gren's syndrome patients. Correlation analysis showed positive correlation between the score of SDS, SAS and the P value (r=0.21, P=0.01) (r=0.14, P=0.007) N dimension (r=0.52, P=0.01) (r=0.55, P=0.007) of EPQ. There were negative correlation between the score of SDS, SAS and the E dimension (r=-0.23, P=0.01) (r=-0.14, P=0.006) of EPQ. Conclusion The prevalence of depres-sion and anxiety is higher in patients with depressive and anxious symptoms. They are in older age, female gender, poor education, longer duration, lower complement and lower T cell subsets when compared with those patients without depression and anxiety. Patients who have an introverted, instable, nervous temperament are more likely to be depressed and anxious. Psychological assessment intervention and therapy should be carried out as early as possible.