Objective To evaluate the role of angiotensin Ⅱ(AngⅡ) signal passway on the expression of Rho GDP dissociation inhibitor alpha (RhoGDIα) in hypertensive rats. Methods Protein and mRNA expressions of RhoGDIα in aortae of 4, 12 and 18 week-old spontaneously hypertensive rats (SHR, n = 4) and Wistar Kyoto rats (WKY, n= 4) were examined by Western blotting and real-time PCR. Aortas from SHR and WKY were analyzed using immonuchemical staining to locate the RhoGDIα in the aorta. The RhoGDIα expression in aorta of hypertensive rat model of aorta coarctation (ACR, n = 6) was also analyzed using Western blotting. Furthermore, The effect of mechanical strain at 10 % elongation on expression of RhoGDIα in vascular smoothmuscle cells (VSMCs) in the presence or absence of L-158809, an antagonist for AngⅡ type 1 receptor, was also evaluated by Western blotting. Results No significant difference of RhoGDIα expression was found between SHR and WKY at 4-week-old and 12-week-old. However, in 18-week-old group, RhoGDIα was significantly highly expressed in SHR than that of WKY at both mRNA and protein levels. RhoGDIα was located in the media of the aorta. Expression of RhoGDIα protein was upregulated in aortas of ACR at 2 and 4 weeks as compared with the controls. The expression of RhoGDIα in VSMCs was inhibited by mechanicalstrain at 10 % elongation, and further decreased by treatment of L-158809. Conclusion RhoGDIα is upregulated in aortae of the hypertensive rats. AngⅡ signal passway may be involved in the process of regulating expression of RhoGDIα.

Objective To clone the fragment of hepatitis C virus(HCV) core gene and express it in E.coli.Methods The fragment of HCV core gene(approximate 366bp) was amplified by PCR and inserted into the pMD18-T vector.The cloned HCV core gene,which was confirmed by the digestion with EcoRⅠ/BamHⅠ,was subcloned into the expression vector pBV220 to construct recombinant plasmid pBV220/HCV-C.The expressed gene product was identified by SDS-PAGE and Western blotting.Results The fragment of HCV core gene was expressed successfully after temperature induction and a protein of 14 000 u was resulted.Conclusion Expression of the HCV-C gene in E.coli was achieved,which may be helpful for further studies on characterizations of HCV-C gene.

It analyzed the definition, mechanism, characteristics of Mirror Visual Feedback and summarized the application of mirror visual feedback in recovering upper limb function after stroke patients at home and abroad, so as to provide evidences for the further research in China.

OBJECTIVETo investigate the association between catecholamine-beta-adrenoceptor (beta-AR)-adenosine 3', 5'-monophosphate (cAMP) system and long-term prognosis in patients with chronic heart failure (CHF).

METHODSThe study population comprised 73 patients with CHF (EF: 23% +/- 10%) with a mean follow-up of 3.8 +/- 1.9 years. Plasma levels of norepinephrine (NE) were measured using high performance lipid chromatography, beta-adrenergic receptor density (Bmax) and the content of cAMP in peripheral lymphocytes were calculated using 3H-dihydroalpneolo as ligand and competitive immunoassay, respectively. Deaths due to cardiovascular events within the follow-up period were registered.

RESULTSThe total mortality was 64.7%, 57.4% of which was for cardiogenic (worsening heart failure: 32.4%; sudden death: 25.0%). In the cardiogenic death group, plasma levels of NE and epinephrine (E) (3.74 nmol/L +/- 0.09 nmol/L and 3.17 nmol/L +/- 1.0 nmol/L) and the contents of peripheral lymphocyte cAMP (3.64 pmol/mg protein +/- 1.4 pmol/mg protein) were significantly increased as compared with the survival group (2.68 nmol/L +/- 0.07 nmol/L, 2.41 nmol/L +/- 0.24 nmol/L and 2.73 pmol/mg protein +/- 0.9 pmol/mg protein, respectively, all P < 0.01). In the sudden death group, plasma levels of NE and E (5.01 nmol/L +/- 0.06 nmol/L and 4.13 nmol/L +/- 0.08 nmol/L) were significantly increased as compared with the worsening heart failure group (2.49 nmol/L +/- 0.07 nmol/L and 2.33 nmol/L +/- 0.8 nmol/L, all P < 0.001) and to the survival group (2.68 nmol/L +/- 0.07 nmol/L and 2.41 nmol/L +/- 0.14 nmol/L, all P < 0.01). The incidences of sudden death were 0%, 75%, and 100% (chi(2) = 16.018, P < 0.01) in patients with plasma NE < 2.5 nmol/L, NE 2.5 nmol/L - 4.5 nmol/L, and NE > 4.5 nmol/L, respectively. In the worsening heart failure group, the content of peripheral lymphocyte cAMP (4.46 pmol/mg protein +/- 0.18 pmol/mg protein) was significantly increased compared with the sudden death group (2.39 pmol/mg protein +/- 0.9 pmol/mg protein, P < 0.001) and to the survival group (2.73 pmol/mg protein +/- 1.1 pmol/mg protein, P < 0.001). The worsening heart failure death occurences were 5.0%, 72.2%, and 100% (chi(2) = 14.26, P < 0.01) in patients with a content of peripheral lymphocyte cAMP < 2.5 nmol/L, cAMP 2.5 nmol/L - 4.5 nmol/L, and cAMP > 4.5 nmol/L, respectively. Bmax in peripheral lymphocyte was not significantly different (P > 0.05) among the sudden death, worsening heart failure, and survival groups in CHF patients.

CONCLUSIONSPlasma levels of catecholamine increase significantly, and Bmax and the contents of cAMP in peripheral lymphocytes decrease significantly in patients with CHF. High plasma catecholamine levels may be associated with sudden death, and high intralymphocyte cAMP content may be associated with worsening heart failure in CHF patients.

Adult ; Aged ; Catecholamines ; blood ; Cyclic AMP ; blood ; Death, Sudden, Cardiac ; Female ; Heart Failure ; blood ; mortality ; Humans ; Lymphocytes ; chemistry ; Male ; Middle Aged ; Receptors, Adrenergic, beta ; blood

Objective To evaluate the diagnostic value of circulating tumor cells(CTCs) in prostate cancer (Pca) through studying the relationship between CTCs and Gleason scores and pathological TNM stage in Pca patients. Methods A total of 238 patients including 161 Pca patients as cancer group, 35 male patients with benign prostatic diseases as benign group and 42 male with non-prostate disease as control group, who were treated in our hospital from July 2016 to January 2018,were enrolled. Venous blood of every patient was collected and CTCs were enriched and identified by immunocytochemistry CD45 capturing leukocyte and fluorescence in situ hybridization with chromosome 8 (CEP8-FISH). Cells displaying CD45-/DAPI+/CEP8>2 were characterized as CTCs. One-way ANOVA was used to exam the correlations of the number of CTCs with Gleason scores and pathological TNM stage. Results CTCs ≥2 were detected in 74.53％(120/161) of Pca patients and 20.00％(7/35)of benign prostatic diseases patients and 7.14％(3/42)of control group (χ2=79.605,P<0.05). In group Gleason scores 6, the numbers of CTCs were 2.00 ± 2.42, the ratios of CTCs≥5 and tetraploid were 13.33％ (2/15)and 26.67％(4/15) respectively. In 7 scores group, the results were 3.14±2.68,17.72％(14/79) and 34.18％(27/79)respectively;In 8 scores group, the results were 3.57 ± 2.70, 33.33％(7/21)and 42.86％ (9/21)respectively; In 9 scores group, these three results were 4.65±4.41, 43.48％(20/46) and 45.65％(21/46)respectively. The numbers of CTCs in the≤pT2b (20), pT2c(27), pT3a(19), pT3b(16)and≥pT4(12) groups were 2.25±2.45, 3.56±2.79, 4.05±3.47, 4.69±2.12 and 5.17±3.21 respectively. The ratios of CTCs≥5 were 25.00％(5/20), 25.93％(7/27), 26.32％(5/19), 50.00％(8/16) and 58.33％ (7/12)respectively. The proportions of tetraploid were 20.00％(4/20), 25.93％ (7/27), 31.58％(6/19), 50.00％(8/16) and 58.33％(7/12) respectively. There were significant differences between CTC and Gleason scores (F=3.200, P<0.05)and pathological stage (F=2.673, P<0.05). The ratios of CTCs≥5 increased with the increase of Gleason scores (χ2=11.592, P<0.05). Conclusions The detection of CTCs could be used for the differential diagnosis of Pca and benign prostatic disease. There were notable correlations between the numbers of CTCs and Gleason scores and pathological stage in Pca patients, especially between CTCs≥5 and Gleason scores.

OBJECTIVE:To optimize the purification technology of total saponins from Asparagus cochinchinensis with macro-reticular resin. METHODS:Using content of total saponins from A. cochinchinensis as index,single factor test was used to investi-gate the macroreticular resin model,sampling adsorption time,mass concentration of the column,adsorption capacity,volume frac-tion and the amount of elution solvent,elution rate,and optimize the purification technology. And verification test was conducted. RESULTS:HPD-300 macroreticular resin showed strong absorption and desorption property. The optimal purification technology was that sampling adsorption time was 60 min,mass concentration of sample liquid was 0.1 g/mL,adsorption capacity was 120 mL(15 BV),it was eluded with 60% ethanol solution with 3 BV and elution rate was 4 BV/h. In the verification test,the average desorption rate of total saponins was 68.30%(RSD=0.95%,n=3). CONCLUSIONS:Optimized purification technology is sta-ble,feasible,and can easily separate and purify the total saponins from A. cochinchinensis.

Objective In this study ,the effect of Spirulina kinase(SPK)on hemorheology ,platelet aggregation and cAMP in acute blood stasis rats model were investigated .Methods Forty SD rats were randomly divided into normal control group ,model group ,aspirin group and the high and low dose of SPK groups .All treatments were performed by gavage once a day for 7 days .Sub-cutaneous injection of adrenalin combined with ice water bath were used to establish the acute blood stasis rat model .The whole blood viscosity ,plasma viscosity ,red blood cell hematocrit (Hct) ,erythrocyte aggregation index (EAI) ,red blood cell deformation index(DI)anderythrocyterigidityindex(ERI)weredetectedbyautomaticbloodflow speedanalyzer .Meanwhile,thelevelofcAMP was detected by ELISA method .Rat platelet aggregation induced by ADP and the maximum aggregation rate was measured by tur-bidimetry .Results Results showed that SPK could significantly decrease the whole blood viscosity ,plasma viscosity ,Hct ,EAI and platelet aggregation rate ,increase the level of cAMP compared with model group(P<0 .05) ,but had no effect on DI and ERI .Con-clusion SPK can markedly improve the abnormal changes of hemorheology in acute blood stasis rats and inhibit the platelet aggregation .

The paper reports a rare case of alkaptonuria (AKU) with IgA nephropathy, and analyzes its clinical manifestations, imaging findings, pathological features, gene diagnosis and treatment process, so as to provide reference for the diagnosis and treatment of the disease. The clinical symptoms of the patient were mainly black urine, microscopic hematuria and proteinuria. Renal pathology showed mild mesangial hyperplasia IgA nephropathy, and renal tubular epithelial cytochrome deposition. Genetic analysis indicated that a pathogenic mutation was detected on the AKU-related homogentisate 1, 2-dioxygenase gene possibly associated with the phenotype of the patient. Genetic testing and renal pathology were effective methods to make a definite diagnosis for the case.

Objective:To investigate the mutation of epidermal growth factor receptor (EGFR), the expression of programmed death ligand 1 (PD-L1), cell proliferation-associated antigen (Ki-67) in elderly patients with non-small cell lung cancer (NSCLC), and their correlation with clinical feature such as gender, histological type and TNM stage.Methods:The tissue samples of 340 elderly NSCLC patients with definite histopathological diagnosis were collected from January 2020 to December 2020 in Huadong Hospital Affiliated to Fudan University, including 195 males and 145 females, age between 68.9±6.0 years. Patients were grouped according to clinical features such as gender, histological type and TNM stage. The expression of EGFR mutation, PD-L1 and Ki-67 were detected by Super-ARMS and immunohistochemistry. The correlation between tnem and clinical features was statistically analyzed, and the correlation between EGFR mutation and PD-L1/Ki-67 expression was further analyzed separately.Results:In elderly NSCLC patients′ tissues, the positive rate of EGFR mutation was 48.53% (165/340). L858R and 19del mutations were the most common types, which were 56.36% (93/165), 30.30% (50/165) respectively. The mutation rate of EGFR was higher in women, lung adenocarcinoma, well-differentiated, and low-stage patients, which were 65.52% (95/145), 53.77% (164/305), 56.75% (143/252), 52.53% (135/257) respectively. In addition, the positive rate of PD-L1 expression was higher in elderly patients with non-adenocarcinoma lung cancer and poorly differentiated adenocarcinoma, which were 37.14% (13/35), 24.53% (13/53) respectively. The negative rate of PD-L1 expression was higher in elderly patients with NSCLC in stage Ⅰ+Ⅱ, no lymph node metastasis and weakly positive Ki-67, which were 89.11% (229/257), 87.63% (248/283), 94.71% (197/208) respectively. Correlation analysis showed that EGFR mutation was negatively correlated with the expression of PD-L1 and Ki-67 (PD-L1: r=-0.22, P<0.001; Ki-67: r=-0.32, P<0.001). Conclusion:There is a negatively correlation between EGFR mutation and the expression of PD-L1 and Ki-67 in elderly NSCLC, suggesting that the combined detection of EGFR mutation and PD-L1 expression could provide the basis for precise targeted therapy for elderly NSCLC patients.

Objective:To study the correlation between circulating tumor cells (CTC) and the degree of pathological invasion, recurrence and metastasis of urothelial carcinoma, and so to explore the clinical value of CTC detection in bladder cancer.Methods:A total of 142 patients with urothelial carcinoma in Huadong Hospital Affiliated to Fudan University were enrolled as cancer group from July 2016 to January 2018. According to the degree of tumor invasion, cancer group was divided into the non-muscle-invasive group (49 cases) and the muscle-invasive group(93 cases). In addition, 52 patients with benign urinary tract lesions admitted were selected as the benign group and 56 patients with non-urinary tract diseases and non-tumor as the control group. A total of 3.2 ml of venous anticoagulant blood from each subject was collected. CTC was enriched by negative enrichment using the magnetic beads coated with monoclonal antibody Cluster 45 of differentiation (CD45) to capture and remove white blood cells, and identified by chromosome 8 probe(CEP8) fluorescence in situ hybridization (FISH) technique. CD45-/4′,6′-diamidino-2-phenylindole+/CEP8>2(CD45-/DAPI+/CEP8>2) cells were judged as CTC. SPSS22.0 statistical software was used for statistical analysis.Results:≥2 CTCs/3.2 ml in blood was set as cutoff value. CTC positive rates in bladder cancer group, benign group and control group were 70.42%(100/142), 28.85%(15/52) and 8.93%(5/56), respectively, and there was a significant difference (χ 2=70.496, P=0.000). There was a statistically difference ( U=2 863.5, P=0.011) in the mean count of CTC(2 CTCs/3.2 ml vs 4 CTCs/3.2 ml) between the two groups. The proportion of≥5 CTCs/3.2 ml in the muscle-invasive group was 40.86% (38/93), which was significantly higher than that in the non-muscle-invasive group, 18.37% (9/49) (χ 2=7.330, P=0.007). Cystoscope follow-up of 65 patients treated with transurethral resection of the bladder tumor showed that the recurrence and metastasis rate in patients with≥5 CTCs/3.2 ml was as high as 47.62% (10/21), compared with 11.36% (5/44) of patients with<5 CTCs/3.2 ml (χ 2=10.530, P=0.001). Among 59 patients undergoing radical cystectomy, no significant difference was found in tumor diameter >3 cm, positive surgical margins and positive lymph nodes among all groups according to CTC negative or positive and CTC number ( P>0.05). But the recurrence and metastasis rate of patients with ≥5 CTCs/3.2 ml (59.10%) was significantly higher than that of patients with <5 CTCs/3.2 ml (6/30)(χ 2=8.364, P=0.004). Conclusion:The number of CTC increased with the deepening of tumor invasion; Tumor recurrence and metastasis increased significantly in the patients with ≥5/3.2 ml CTCs in blood.