Aim To investigate the role of endothe-lium in the enhancement of phenylephrine-mediated vasoconstriction by bupivacaine in the isolated rat aor-ta.Methods The isolated rat aortic rings were pre-pared, and the vascular endothelium was removed chemically or physically .Phenylephrine-mediated vas-oconstriction was recorded .Results A pretreatment with bupivacaine at 30 μmol · L-1 for 20 min signifi-cantly increased the Emax value of vasoconstrictive re-sponses to phenylephrine from 2.22 ±0.07 g of sol-vent-controlled group to 2.50 ±0.05 g ( P<0.01 ) in the isolated endothelium-intact rat aorta.However, the Emax value was not significantly changed by a pretreat-ment with bupivacaine at 30 μmol · L-1 for 5 , 10 or 15 min ( P>0.05 ) .A pretreatment with bupivacaine at 30 μmol · L-1 for 20 min slightly but significantly inhibited the vasoconstrictive responses to low concen-tration of phenylephrine in the isolated endothelium-de-nuded rat aorta (P<0.05).In the isolated endotheli-um-intact rat aorta under a combined treatment with in-dometacin, ChTX, apamin and L-NAME, the vasodi-lator responses to acetylcholine were completely sup-pressed , and a pretreatment with bupivacaine at 30μmol· L-1 for 20 min did not significantly affect the vasoconstrictive responses to phenylephrine ( P >0.05 ) .Conclusion Bupivacaine enhances α1-adre-noceptor-mediated vasoconstriction by inhibiting vascu-lar endothelium in the isolated endothelium-intact rat aorta, Which potentiates indirectly the vasoconstrictive responses to phenylephrine .

Aim To investigate the selective inhibition of ethanol on muscarinic receptor-or 5-HT receptor-me-diated contractile responses in the circular smooth mus-cle strips isolated from the different regions of rat stom-ach. Methods Circular muscle strips isolated from the rat gastric fundus, body, cardia and pylorus were prepared, and the contractile responses to carbachol ( CCh ) or 5-HT were recorded. Results Ethanol (0. 000 05~0. 000 5, V/V) did not affect the contrac-tile response to CCh in circular muscle strips from the rat gastric fundus and cardia, and that to 5-HT in the strips from rat gastric fundus and body ( P >0. 05 ) . However, ethanol(0. 000 1 and 0. 000 5) significantly inhibited the Emax value of the contraction by CCh from (12. 18 ± 0. 33) g of control level to (10. 88 ± 0. 41) g and -lgEC50 value from ( 6. 33 ± 0. 05 ) of control level to (6. 12 ± 0. 06)(P <0. 05) in the strips from rat gastric body. Ethanol(0. 000 1 and 0. 000 5) also significantly inhibited the Emax value of the contraction by CCh from (2. 87 ± 0. 15) g of control level to (2. 2 ± 0. 13) g and -lgEC50 value from (6. 49 ± 0. 10) of control level to (6. 05 ± 0. 09)(P<0. 01) in the strips from rat gastric pylorus. Moreover, ethanol ( 0. 000 1 and 0. 000 5) significantly inhibited the Emax value of the contraction by 5-HT from (2. 93 ± 0. 35) g of con-trol level to ( 2. 1 ± 0. 30 ) g ( P<0. 05 ) , but did not affect the -lgEC50 value in the strips from rat gastric cardia. Conclusions Ethanol inhibits the contractile responses to 5-HT only in the circular muscle strips of rat gastric cardia, and it inhibits the contractile respon-ses to CCh more strongly in the circular muscle strips of gastric pylorus than gastric body. In those gastric circular muscle strips, ethanol decreases both the ac-tivity and affinity of CCh to muscarinic receptors, but decreases only the activity of 5-HT to its receptors.

Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.

Objective: To describe the clinical characteristics with long-term prognosis in patients with mid-ventricular obstructive hypertrophic cardiomyopathy (MVOHCM).
Methods: A total of 66 MVOHCM patients treated in our hospital were retrospectively studied for their morbidity, clinical characteristics and mortality. The cumulative survival rate was calculated by Kaplan-Meier method; the risk factors for cardiac death and cardiovascular events were analyzed by uni- and multivariate Cox proportional hazard model.
Results: There were 66 (2.74%) patients suffering from MVOHCM among 2413 patients of hypertrophic cardiomyopathy and the average diagnostic age was (40.16 ± 14.64) years. With (7.30 ± 6.25) years of follow-up study, the cardiovascular mortality was 13.6% and unexplained syncope (HR=13.37, 95% CI: 1.65-114.46, P=0.015) was the independent predictor for cardiovascular death. There were 45.45% (30/66) patients experienced at least 1 time of cardiovascular event and the most frequent one was non-sustained ventricular tachycardia (NSVT); 19.70% (13/66) of patients combined with apical aneurysms, and they were more inclined to experience NSVT.
Conclusion: MVOHCM patients usually have unfavorable prognosis with the higher incidence of cardiovascular events, some patients may develop apical aneurysm. The early diagnosis of MVOHCM is important for appropriate treatment.

Background: The dietary agent sulforaphane (SFN) has been reported to reduce diabetes-induced renal fibrosis, as well as inhibit histone deacetylase (HDAC) activity. Bone morphologic protein 7 (BMP-7) has been shown to reduce renal fibrosis induced by transforming growth factor-beta1. The aim of this study was to investigate the epigenetic effect of SFN on BMP-7 expression in diabetes-induced renal fibrosis. Methods: Streptozotocin (STZ)-induced diabetic mice and age-matched controls were subcutaneously injected with SFN or vehicle for 4 months to measure the in vivo effects of SFN on the kidneys. The human renal proximal tubular (HK11) cell line was used to mimic diabetic conditions in vitro. HK11 cells were transfected to over-express HDAC2 and treated with high glucose/palmitate (HG/Pal) to explore the epigenetic modulation of BMP-7 in SFN-mediated protection against HG/Pal-induced renal fibrosis. Results: SFN significantly attenuated diabetes-induced renal fibrosis in vivo. Among all of the HDACs we detected, HDAC2 activity was markedly elevated in the STZ-induced diabetic kidneys and HG/Pal-treated HK11 cells. SFN inhibited the diabetes-induced increase in HDAC2 activity which was associated with histone acetylation and transcriptional activation of the BMP-7 promoter. HDAC2 over-expression reduced BMP-7 expression and abolished the SFN-mediated protection against HG/Pal-induced fibrosis in vitro. Conclusion Our study demonstrates that the HDAC inhibitor SFN protects against diabetes-induced renal fibrosis through epigenetic up-regulation of BMP-7.

OBJECTIVETo assess the pathologic markers for evaluation of reversibility in pulmonary hypertension (PAH) related to congenital heart disease.

METHODSTwenty-eight patients with congenital heart disease complicated by PAH were subclassified into reversible pulmonary hypertension (RPAH) and irreversible pulmonary hypertension (IPAH), according to post-operative mean pulmonary artery pressure (MPAP). Pulmonary vascular lesion was analyzed according to Ruan's method. Mean medium thickness percent, mean medium area percent and pulmonary arteriolar density were measured by quantitative morphometry. Immunohistochemical study for transgelin and filamin A was carried out.

RESULTSAmongst the 28 cases studied, 24 were RPAH and 4 were IPAH. Of the 24 patients with RPAH, 13 (54.2%, 13/24) had pulmonary vascular lesion of grade 0, 9 (37.5%, 9/24) of grade 1 and 2 (8.3%, 2/24) of grade 2. Of the 4 patients with IPAH, 1 had lesion of grade 1, 1 of grade 2 and 2 of grade 3. Both preoperative and postoperative MPAP were higher in IPAH patients than that in RPAH patients[(53.3±23.4) mmHg versus (34.1±12.7) mmHg, P=0.020 and (35.0±8.8) mmHg versus (17.8±3.9) mmHg, P<0.01]. Compared to patients with pulmonary vascular lesion of grades 0 and 1, the preoperative MPAP in patients with grades 2 and 3 showed no significant difference, but the postoperative MPAP was higher (P<0.05 or 0.01). Compared to control group, mean medium thickness percent and mean medium area percent were significantly higher in RPAH and IPAH categories (12.0±3.5, 8.5±2.0 versus 5.7±1.0, P<0.01 and 55.8±11.1, 49.0±9.4 versus 34.0±5.5, P<0.01). Mean medium thickness percent was significantly higher in IPAP group than that in RPAH group (12.0±3.5 versus 8.5±2.0, P=0.001). Correlation analysis demonstrated that mean medium thickness percent and mean medium area percent had positive correlation with preoperative and postoperative MPAP. There was no correlation between grading of pulmonary vascular lesion and reversibility. Transgelin and filamin A had stronger staining in pulmonary vascular smooth muscle cells in IPAH than those in RPAH and controls(P<0.05).

CONCLUSIONSPathologic assessment of lung biopsy remains the gold standard for evaluation of the reversibility in PAH related to congenital heart disease. Mean medium thickness percent, mean medium area percent and immunoreactivity for transgelin and filamin A are useful parameters.

Biomarkers ; metabolism ; Biopsy ; Filamins ; metabolism ; Heart Diseases ; complications ; pathology ; Humans ; Hypertension, Pulmonary ; complications ; diagnosis ; pathology ; Lung ; pathology ; Microfilament Proteins ; metabolism ; Muscle Proteins ; metabolism

Objective:To construct and validate a nomogram model based on clinical factors and PET/CT metabolic parameters of 18F-FDG for predicting epidermal growth factor receptor (EGFR) mutations in lung adenocarcinoma. Methods:From January 2014 to January 2019, 114 patients (59 males, 55 females, age (60.0±10.8) years) with lung adenocarcinoma in the First Affiliated Hospital of Harbin Medical University were retrospectively enrolled. Clinical data (smoking status, tumor location, clinical stage and carcinoembryonic antigen (CEA) level), 18F-FDG PET/CT metabolic parameters (SUV max, metabolic tumor volume (MTV) and total lesion glycolysis (TLG)) and EGFR mutation status were analyzed. Patients were divided into training group (80 cases) and validation group (34 cases). In the training group, univariate analyses (independent-sample t test, Wilcoxon rank sum test, χ2 test or Fisher′s exact probability method) were used for categorical variables. Variables that showed significant differences between EGFR mutation group and wild type group were selected. Variance inflation factors (VIF) were calculated and the collinearity variables were deleted, and a nomogram model of optimal logistic model was constructed based on Akaike information criterion (AIC). The effect of the model was evaluated by the concordance index (C-index), sensitivity, specificity, accuracy, calibration and decision curve analysis (DCA) in the training group and the validation group. Results:Among 114 patients, 56 were with EGFR mutations and 58 were with EGFR wild type. In the training group, there were significant differences in gender (male/female: 14/26 vs 25/15; χ2=6.05, P=0.014), smoking status (with/without smoking history: 4/36 vs 22/18; χ2=18.46, P<0.001) and SUV max (5.72(3.90, 8.32) vs 8.09(4.56, 12.55); W=1 045.50, P=0.018) between EGFR mutation group and wild type group. However, there were no significant differences in other factors ( t=-0.54, χ2 values: 0.20 and 0.20, W values: 921.50 and 983.00, all P>0.05). The VIF of gender, smoking status and SUV max were all less than 10, and the nomogram model with three factors showed the minimum AIC (90.06). In the training group, C-index value of the model was 0.798 (95% CI: 0.699-0.897), with the sensitivity of 85.0%(34/40), the specificity of 70.0%(28/40) and the accuracy of 77.5%(62/80). In the validation group, C-index value was 0.854(95% CI: 0.725-0.984), with the sensitivity of 13/16, the specificity of 14/18, and the accuracy of 79.4%(27/34). The calibration curve and the goodness of fit test showed good calibration, and DCA showed that the model could benefit patients clinically within a large risk threshold range (training group: 0-0.59, validation group: 0-0.65). Conclusion:The nomogram model based on gender, smoking status and SUV max can be used to easily predict EGFR mutation status in lung adenocarcinoma.

Objective:To investigate the clinical effects of tibial transverse bone transport assisted by nose ring drainage (NRD) in the treatment of foot and ankle chronic osteomyelitis.Methods:A retrospective study was conducted to analyze the data of 32 patients with foot and ankle chronic osteomyelitis who had been treated at Department of Orthopedics, Beijing Rehabilitation Hospital from March 2013 to February 2022. The patients were assigned into a study group and a control group. In the control group, there were 12 males and 3 females, aged (39.5±8.8) years. The osteomyelitis was located at the distal tibia in 4 cases, at the talus in 1 case, at the calcaneus in 2 cases, at the midfoot in 4 cases and at the forefoot in 4 cases. According to the Crerny-Mader classification, there were 5 cases of type Ⅰ and 10 cases of type Ⅱ. The control group was treated with Ilizarov transverse tibial bone transport in combination with thorough debridement and anti-infection therapy. In the study group, there were 12 males and 5 females, aged (42.3±13.4) years. The osteomyelitis was located at the distal tibia in 4 cases, at the talus in 1 case, at the calcaneus in 3 cases, at the midfoot in 5 cases and at the forefoot in 4 cases. According to the Crerny-Mader classification, there were 7 cases of type Ⅰ and 10 cases of type Ⅱ. The study group was treated with NRD drainage in addition to the procedures in the control group. The curative effects were evaluated by comparing the cure, recurrence, amputation (toe amputation), antibiotic use time, wound healing time, relevant inflammatory indicators [WBC, neutrophil percentage (NEU), erythrocyte sedimentation rate (ESR), procalcitonin, interleukin-6 (IL-6), C-reactive protein (CRP)], and postoperative functional recovery between the 2 groups.Results:The 2 groups were comparable because there was no statistically significant difference in the general data between them ( P>0.05). The follow-up period was (36.0±9.8) months. There was no significant difference between the 2 groups in the cure or amputation (toe amputation) ( P>0.05), but the recurrence rate in the study group was significantly lower than that in the control group ( P<0.05). The antibiotic use time [(20.7±3.6) d] and wound healing time [(88.3±17.1) d] in the study group were significantly shorter than those [(37.9±6.5) d and (102.2±22.6) d] in the control group ( P<0.05). The ESR, IL-6 and CRP at 1 week after operation in the study group were significantly lower than those in the control group ( P<0.05), but there was no significant difference between the 2 groups in the WBC, NEU or PCT at 1 week after operation ( P>0.05). There was no significant difference either in the inflammatory indicators between the 2 groups at 1 month after operation ( P>0.05). In all patients, the inflammatory indicators like WBC, NEU, ESR, PCT, IL-6 and CRP at 1 week and 1 month after operation were significantly better than those before operation, and the Maryland foot functional score at 12 months after operation was significantly higher than the preoperative one ( P<0.05). There was no significant difference between the 2 groups in the Maryland foot functional score at 12 months after operation ( P>0.05). Conclusion:In the treatment of foot and ankle chronic osteomyelitis, compared with the Ilizarov transverse tibial bone transport, our tibial transverse bone transport assisted by NRD can achieve satisfactory therapeutic effects, shorten antibiotic use time and treatment cycle, and reduce recurrence rate.

OBJECTIVETo investigate the histopathological features of primary restrictive cardiomyopathy (PRCM).

METHODSNine extransplanted hearts from heart transplantation recipients were examined. Gross and histopathological findings were observed, photographed and final pathological diagnosis was compared to clinical diagnosis. The myocardial ultrastructure changes were determined using transmission electron microscopy.

RESULTSThe hallmark pathologic feature of PRCM was distinguished by myocardial cell degeneration and hyperplastic collagen fibrils around the myocardial cells.Fibrosis was severer in left ventricle free wall than in ventricular septum and right ventricle. The degree of myocardial cell degeneration and poloidal disorder were severer in patients with reduced ejection fraction (EF) than in patients with preserved EF. Transmission electron microscope evidenced severe interstitial fibrosis, myofibrillar changes of sarcomere structure, abnormalities both on intercalated disc number and distribution.

CONCLUSIONSPRCM is characterized by hyperplastic collagen fibrils around the cardiomyocytes. Fibrosis is severer in left ventricle than in right ventricle. Sarcomere dysplasia is the main cause of PRCM, and ultrastructural examination is helpful for PRCM diagnosis.

Cardiomyopathy, Restrictive ; surgery ; Fibrosis ; Heart Transplantation ; Heart Ventricles ; Humans ; Myocardium ; pathology ; Myocytes, Cardiac ; Sarcomeres

OBJECTIVETo study the clinical and pathologic features of primary cardiac inflammatory myofibroblastic tumor.

METHODSA total of 4 patients with primary cardiac inflammatory myofibroblastic tumor were encountered during the period from 1993 to 2013 in National Center for Cardiovascular Disease. The clinical features, imaging findings and outcomes of the 4 patients were evaluated. ALK protein expression and ALK gene status were studied using the archival tumor tissues.

RESULTSThere were 1 female and 3 male patients. The age of patients ranged from 5 months to 30 years (mean = 16 years). The tumor was located in right ventricle (n = 2), right atrium (n = 1) or pericardium (n = 1). Histologic patterns included 2 cases of fibrous histiocytoma type, 1 case of granulomatous type and 1 case of sclerosing type. Immunohistochemical study showed that 2 cases expressed ALK protein. Fluorescence in-situ hybridization however did not reveal any ALK gene rearrangement.

CONCLUSIONSInflammatory myofibroblastic tumor of the heart is rarely encountered and easily misdiagnosed. It carries distinctive clinical and pathologic features. ALK protein expression is helpful in arriving at the correct diagnosis.

Adolescent ; Adult ; Biomarkers, Tumor ; genetics ; metabolism ; Child ; Diagnosis, Differential ; Female ; Granuloma, Plasma Cell ; enzymology ; pathology ; Heart Neoplasms ; enzymology ; pathology ; Histiocytoma, Benign Fibrous ; enzymology ; pathology ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Infant ; Male ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism