Value of CHCHD2 as a potential marker of non-small cell lung cancer: analysis of 60 cases / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of CHCHD2, a potential tumor marker, in tumor and adjacent tissues from patients with non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Immunohistochemistry was used to detect the expression and location of CHCHD2 in the tumor tissues from 60 patients with NSCLC and 35 adjacent tissues to analyze the correlation of CHCHD2 expression with the clinicopathological variables and overall survival of the patients. The expression profile of CHCHD2 mRNA in NSCLC was analyzed using Oncomine database.</p><p><b>RESULTS</b>The positivity rate of CHCHD2 was significantly higher in the tumor tissues than in adjacent tissues in patients with NSCLC (75.0% vs 17.1%). CHCHD2 positivity in the tumor tissues was associated with lymph node metastasis, pathological TNM stage, and tumor grades but not with age, gender, or histological type of the tumors. Analysis using Oncomine database showed that CHCHD2 mRNA was expressed at significantly higher levels in NSCLC than in normal control group (P<0.05). Kaplan-Meier survival analysis showed that NSCLC patients with a positive expression of CHCHD2 had a significantly shorter overall survival time than those negative for CHCHD2 (P<0.05).</p><p><b>CONCLUSION</b>As a potential tumor marker, CHCHD2 over-expression plays a role in the occurrence and progression of NSCLC and promotes tumor invasion and metastasis, and can potentially serve as an indicator for early diagnosis and prognostic evaluation of NSCLC.</p>

Establishing a rat model of intervertebral disc degeneration using three methods / 中国组织工程研究

BACKGROUND: Numerous studies focus on animal models of intervertebral disc degeneration (IDD), but criteria for establishing the animal models of IDD have not been confirmed, and there is a lack of systematic comparison among models. OBJECTIVE: To compare the rat models of IDD established by puncturing at annulus, endplate injection and their combination, thus providing reference for IDD model selection. METHODS: Eighty Sprague-Dawley rats were equivalently randomized into four groups: puncturing group (puncturing at the annulus), endplate injection group (endplate injected with ethyl alcohol), combination group (puncturing at the L5-6annulus and endplate injection at the same segment) and sham operation group. Three rats in each group were taken at postoperative 4, 8, and 12 weeks for X-ray examination to measure the disc height; and the discs were removed for histological observation and immunohistochemical staining. RESULTS AND CONCLUSION: The results of X-ray examination, hematoxylin-eosin staining and immunohistochemical staining all showed that the IDD degree was gradually aggravated in all groups except the sham operation group. At postoperative 4 weeks, compared with the sham operation group, in the endplate injection and combination groups, the percent disc height was significantly decreased, the pathological scores were significantly increased and the average gray value of collagen type I was significantly reduced (P ＜ 0.05). At postoperative 8 and 12 weeks, compared with the sham operation group, the percent disc height in the other three groups were all significantly decreased, the pathological score was significantly increased, and the average gray value of collagen type I was significantly decreased (P ＜ 0.05). Compared with the puncturing and endplate injection group, in the combination group, the percent disc height at postoperative 8 weeks was significantly decreased, and the average gray value of collagen type I at postoperative 12 weeks was significantly decreased (P ＜ 0.05). These results suggest that the rat IDD model can be successfully constructed by above three methods. Puncturing at the annulus is easy to operate and control IDD progression, which can be used to study different stages of IDD. Endplate injection is suitable for the etiological study of IDD, and induces IDD earlier than puncturing, but the final results are similar. The combination method can significantly accelerate IDD aggravation, and thus is not time consuming.

Influence of bear bile on rat hepatocarcinoma induced by diethylnitrosamine / 药学学报

To investigate the influence of bear bile on rat hepatocarcinoma induced by diethylnitrosamine (DEN), a total of 40 rats were randomly divided into 4 groups: normal control group, model group, and two bear bile treatment groups. The rat liver cancer model was induced by breeding with water containing 100 mg x L(-1) DEN for 14 weeks. The rats of the bear bile groups received bear bile powder (200 or 400 mg x kg(-1)) orally 5 times per week for 18 weeks. The general condition and the body weight of rats were examined every day. After 18 weeks the activities of serum alanine transaminase (ALT), aspartate transaminase (AST) and total bilirubin (TBIL) were detected. Meanwhile, the pathological changes of liver tissues were observed after H&E staining. The expression of proliferative cell nuclear antigen (PCNA) and a-smooth muscle actin (alpha-SMA) in liver tissue were detected by immunohistochemical method. After 4 weeks the body weights of rats in normal group were significantly more than that in other groups (P < 0.05); and that in the two bile groups was significantly more than that in the model group. Compared with normal group, the level of serum glutamic-pyruvic transaminase and total bilirubin increased significantly in other groups; compared with model group, these two indexes decreased significantly in two bile groups. Hepatocellular carcinoma occurred in all rats except for normal group; there were classic cirrhosis and cancer in model group while there were mild cirrhosis and high differentiation in two bile groups. There were almost no expressions of PCNA and alpha-SMA in normal group while there were high expressions in model group; the two bile groups had some expressions but were inferior to the model group, and alpha-SMA reduced markedly. It indicated that bear bile restrained the development of liver cancer during DEN inducing rat hepatocarcinoma, which may be related to its depressing hepatic stellate cell activation and relieving hepatic lesion and cirrhosis.

Humanized monoclonal antibody TNT-3-mediated truncated tissue factor for the treatment of H22 hepatoma-bearing mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the inhibitory effects of humanized monoclonal antibody-3 (huTNT-3) mediated truncated tissue factor (tTF) on the H(22) hepatoma-bearing mice, and to explore its mechanisms.</p><p><b>METHODS</b>The coagulation activity of the huTNT-3/tTF fusion protein was detected by clotting assay and clotting factor X (FX) activation test in vitro. Mouse hepatoma cell line H(22) cells were inoculated subcutaneously into mice to establish the mouse models of hepatoma. The mice were randomly divided into two groups to be injected once with huTNT-3/tTF fusion protein or tTF protein labeled with rhodamine B isothiocyanate (RBITC), respectively. The localization of huTNT-3/tTF fusion protein in the mouse hepatoma tissue was analyzed by confocal laser scanning microscopy 24 hour after the injection. Fifteen mice were randomly divided into three groups to be injected with the huTNT-3/tTF fusion protein, tTF protein or phosphate buffered saline (PBS) once, respectively. The tumor size was measured every two days to calculate the tumor volume. Ten days after the injection the mice were sacrificed. Samples of the tumor, heart, livers, spleen, lung, kidney and brains of the mice were taken for histopathological examination.</p><p><b>RESULTS</b>Both the huTNT-3/tTF fusion protein and tTF protein effectively promoted blood coagulation. Under the conditions of Ca(2+), the coagulation time in the 1.5, 3, 6 µmol/L huTNT-3/tTF groups was (12.90 ± 0.60) min, (10.39 ± 0.40) min and(8.15 ± 0.24) min, respectively, and the coagulation time of the 1.5, 3, 6 µmol/L tTF groups was (14.23 ± 0.46) min, (12.10 ± 0.49) min and (9.83 ± 0.52) min, respectively, the difference between the two groups was not significant (F = 0.145, P = 0.705). The huTNT-3/tTF fusion protein was similar to the tTF protein in the ability of activating FX (t = 0.101, P > 0.05). The confocal laser scanning microscopic analysis showed that RBITC-fluorescence labeled huTNT-3/tTF fusion protein was enriched in the hepatoma tissue. The tumor volume of the huTNT-3/tTF fusion protein group was significantly lower than that of the tTF and PBS groups (both P < 0.001), however, there was not significant difference between the tTF and PBS groups (t = -0.616, P > 0.05). The survival time of the huTNT-3/tTF group was (25.5 ± 2.5) d, significantly longer than that of the PBS group (17.3 ± 1.9) d and the tTF group (18.6 ± 1.9) d, (both P < 0.05).</p><p><b>CONCLUSION</b>The huTNT-3/tTF fusion protein retains the coagulation ability and has the capability of targeting to tumor vasculature, and induces thrombosis in the tumor vessels, thus to suppress the growth of hepatoma in the mice.</p>

Pharmacokinetics of photosensitizer m-THPC in rat models of liver cancer via orthotropic implantation using Walker-256 / 中华外科杂志

<p><b>OBJECTIVE</b>To study the pharmacokinetics, distribution and excretion of m-THPC in rat models of liver cancer via orthotropic implantation using Walker-256.</p><p><b>METHODS</b>After an intravenous injection of m-THPC with 0.3 mg/kg, the concentrations of m-THPC in biological specimens were determined by a fluorescence method. The data obtained were processed with PK-GRAPH pharmacokinetic procedure.</p><p><b>RESULTS</b>The disposition of m-THPC in rat models of liver cancer Walker-256 was conformed to a two compartment model with T(1/2)α = 1.18 h, T(1/2)β = 22.57 h at the dose of 0.3 mg/kg.m-THPC was shown to be widely distributed to the various tissues. There was a highest drug accumulation in liver and liver cancer, and lowest in skin and muscle. Ratio of m-THPC concentration in the Walker-256 tumor compared to normal tissue reach the peak 24 h after m-THPC administration.</p><p><b>CONCLUSIONS</b>m-THPC is distributed widely and eliminated at a rapid rate in Walker-256 rats. Twenty four hours after m-THPC administration may be the best time for photodynamic therapy of liver cancer.</p>

Comparisons of endocrine hormones levels between Tibetan antelope and Tibetan sheep / 生理学报

The Tibetan antelope, a prototype mammal, has developed a unique adaptation to extreme high altitude-associated hypoxia. To investigate the role of the endocrine system in adaptation to high altitude in the Tibetan antelope, comparisons of endocrine hormones levels between Tibetan antelope (n = 9) and Tibetan sheep (n = 10) were performed. Both two kinds of animals were captured at an altitude of 4 300 m and then transported to experimental base at 2 800 m altitude. The blood samples were drawn from right external jugular vein in the next morning, and the 20 hormones in hypothalamus-adenohypophysis-peripheral hormonal axis were measured with radioimmunoassay or enzyme-linked immunosorbent assay. Heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean pulmonary arterial pressure (mPAP) were recorded using catheterization. Moreover, hemoglobin (Hb) content was measured by blood analyser. The results showed that, the levels of FT(3), FT(4) and Ang II in Tibetan antelope were significantly lower than those in Tibetan sheep, whereas TRH, CRH, GHRH, F, E(2), Ald, ACTH and CGRP levels were significantly greater in Tibetan antelope than those in the Tibetan sheep. Compared with Tibetan sheep, Tibetan antelope showed lower HR, mPAP, SBP, DBP and Hb content. In Tibetan antelope and Tibetan sheep, both Hb and Ang II were correlated positively with respective mPAP. In Tibetan antelope, FT(3) level was correlated positively with GH and negatively with ACTH. These results suggest that the endocrine system of Tibetan antelope is characterized by low energy expenditure and high stress, which may be one of the mechanisms underlying the Tibetan antelope adaptation to chronic hypoxia.

Arginine-glycine-aspartic polypeptide mediated truncated tissue factor therapy for colorectal carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the therapy effects of (arginine-glycine-aspartic, RGD)(3)-truncated tissue factor (tTF) fusion protein on colorectal carcinoma in mice.</p><p><b>METHODS</b>The (RGD)(3)-tTF fusion gene, constructed with tTF and three series-wound peptides RGD, was expressed in Escherichia coli BL21 (DE(3)). The fusion protein was purified through Nickel affinity chromatography column. The coagulation activity of the (RGD)(3)-tTF fusion protein was detected by clotting assay in vitro. Mice colorectal cancer cells line CT26 were inoculated subcutaneously into mice to establish colorectal cancer model. Four mice were randomly divided into two groups to be injected with the (RGD)(3)-tTF or tTF fusion protein labeled with rhodamine B isothiocyanate (RBITC) at a single dose of 50 microg respectively. The location of the (RGD)(3)-tTF fusion protein in the colorectal carcinoma bearing mice tissue was analyzed by using in vivo optical imaging one hour after the injection and confocal microscopy twenty-four hours after the injection. Fifteen mice bearing colorectal carcinoma were randomly divided into three groups for injection with the (RGD)(3)-tTF, tTF fusion protein or phosphate buffered saline (PBS) at a single dose of 50 microg respectively. The tumor size was measured daily to calculate the tumor volume. Five days after the injection, the mice were killed to harvest tumor tissues, hearts, livers, spleens, lung, kidneys and brains to observe valid thrombogenesis and tumor necrosis.</p><p><b>RESULTS</b>With the concentration of the (RGD)(3)-tTF fusion protein increased, the clotting time was shorten correspondingly under the conditions of Ca(2+), and the clotting time was (8.6 +/- 0.2) min when the concentration was 6 micromol/L, and it was >30 min in the group of 0 micromol/L (P < 0.05). The coagulation activity of (RGD)(3)-tTF and tTF fusion protein was alike (F = 0.09, P > 0.05). The in vivo optical imaging and confocal microscopy analyses showed that RBITC fluorescence labeling (RGD)(3)-tTF fusion protein was assembled in the tumor vasculature. On the first, third, fifth day after injection, the tumor volume of (RGD)(3)-tTF fusion protein group was (120.8 +/- 4.8) mm(3), (93.8 +/- 3.4) mm(3), (132.2 +/- 7.7) mm(3) respectively, which was significantly smaller than that of the tTF group [(181.4 +/- 13.8) mm(3), (333.0 +/- 32.0) mm(3), (514.0 +/- 11.5) mm(3)] and PBS group [(182.6 +/- 11.5) mm(3), (332.8 +/- 21.0) mm(3), (524.2 +/- 16.7) mm(3)] (both P < 0.05). However, there was no significant difference in the tumor volume between the latter two groups (P > 0.05).</p><p><b>CONCLUSION</b>The (RGD)(3)-tTF fusion protein is capable of targeting to tumor vasculature and inducing thrombogenesis for suppressing the tumor growth in the colorectal carcinoma mice model, and it's expected to be a new therapy for colorectal cancer.</p>

Clinical outcome of mitral valve repair in primary infective endocarditis with mitral insufficiency / 中华外科杂志

<p><b>OBJECTIVE</b>To study the clinical results of mitral valve repair in patients of primary infective endocarditis with mitral insufficiency.</p><p><b>METHODS</b>From January 2004 to July 2007, 40 patients who had undergone valve repair procedure for infective endocarditis with mitral insufficiency were analyzed retrospectively. There were 26 male and 14 female patients, with an average age of (34.0 +/- 3.5) years old, including 6 patients of underlying heart disease, 34 patients of no previously underlying heart disease. There were 12 patients in NYHA functional class II, 19 patients in class III, 9 patients in class IV preoperatively. Preoperative echocardiography showed moderate to severe MR in all patients. The surgery was performed under extracorporeal circulation and moderate hypothermia. The distribution of anatomical lesion according to surgical findings were vegetation in 32 patients, leaflet prolapsed in 34 patients, leaflet perforation in 16 patients, chordal rupture in 32 patients, and annular abscess in 2 patients. The vegetations and infected tissues were debrided. The surgery consisted of complex methods to repair mitral valve, including direction leaflet closure in 5 patients, pericardial patch closure of leaflet perforation in 18 patients, chords reimplantation in 4 patients and chords transference in 6 patients, quadrangular resection in 12 patients, double orifice method in 17, closure of the commissure in 8, rings annuloplasty in 28 cases, and so on. There were 28 selective surgeries and 12 emergent ones. Patients were evaluated for early and long-term clinic and echocardiographic outcome before and after operation.</p><p><b>RESULTS</b>There were no early postoperative death. Mitral valve repair was feasible in 39 patients, one patient was transformed to mitral valve replacement during the operation. Postoperative echocardiography demonstrated no (n = 24) or mild (n = 15) mitral regurgitation at the discharge examination and observed significant reductions in left ventricular end diastolic [from (62 +/- 7) mm to (51 +/- 6) mm, P < 0.05] and end systolic dimensions [from (45 +/- 3) mm to (40 +/- 4) mm, P < 0.05] and left atrial dimensions [from (49 +/- 4) mm to (42 +/- 6) mm, P < 0.05]. Mean follow-up (25.6 +/- 3.2) months, freedom from recurrent moderate to severe MR, freedom from repeat operation or infective endocarditis, revealed patients were 36 cases in NYHY class I, 3 cases in class II.</p><p><b>CONCLUSION</b>Mitral valve repair for mitral valve endocarditis is feasible with a satisfied clinical outcome, maintains valve competency with significant reductions in left atrial and left ventricular dimensions after surgery.</p>

Histone deacetylase inhibitor SAHA induces inactivation of MAPK signaling and apoptosis in HL-60 cells / 中国实验血液学杂志

The study was aimed to investigate the molecular mechanisms of histone deacetylase inhibitor SAHA-induced apoptosis of acute myeloid leukemia cell line HL-60. The effect of SAHA on HL-60 cell proliferation was detected by MTT assay and the cell morphological changes were observed with Wright-Giemsa and Hoechst33342 staining. The cell cycle distribution was determined by flow cytometry and the expression of cell signaling proteins were detected by Western-blot analysis. The results showed that SAHA inhibited the proliferation of HL-60 cells in dose- and time-dependent manners, after 2 micromol/L SAHA exposure for 12 - 48 hours, the cell cycle was arrested at G(0)/G(1) phase and apoptotic cell death was confirmed by either defined apoptotic bodies stained by Hoechst33342, Western blot showed cleaved-PARP, which represents the activation of caspase 3. The Western blot analysis indicated the activation of two important survival signal pathways after SAHA treatment, the phosphorylation of Raf and its downstream ERK kinases were remarkable downregulated, whereas the phosphorylation of AKT and its downstream molecular mTOR were not changed. It is concluded that SAHA-induced apoptosis of HL-60 cells is mediated by inactivation of p44/42 MAPK signaling pathway.

Comparative analysis of the risk factor on the clinical information of myocardial revascularization combined with concomitant valve operations / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the clinic information of coronary artery bypass grafting (CABG) combined with concomitant valve operation.</p><p><b>METHODS</b>Retrospectively analyze the information of morbidity and mortality of 126 cases patients who underwent combined valve and bypass procedures between December 2000 and January 2005. These patients had been divided into 2 groups according to sex.</p><p><b>RESULTS</b>There were significant differences in the clinic characteristic such as weight and diabetes mellitus and mitral valve stenosis and three disease vessels of coronary artery between 2 groups (P < 0.05). The rate of the number of bypass grafts and morbidity and mortality of complication were significant differences (P < 0.05). The number of mitral valve replacement of female was more than that of male (P < 0.05). Five males died after operation, 1 case of heart failure, 1 case of high blood sugar, 2 cases of arrhythmia, 2 cases of organs failure; Seven females died after operation, one case of heart failure, one case of alimentary tract haemorrhage, three cases of arrhythmia, two cases of organs failure.</p><p><b>CONCLUSIONS</b>Coronary artery bypass grafting (CABG) combined with concomitant valve operation is safe and effective. The rate of morbidity and mortality of complication of female is more than that of male. The study demonstrates that female gender is an independent risk factor for combined morbidity and mortality during and after combined valve and coronary bypass surgery. That is related to low weight and mitral valve stenosis of female.</p>

Off-pump coronary artery bypass grafting with only bilateral internal mammary artery composite Lima-Rima Y graft / 中华外科杂志

<p><b>OBJECTIVE</b>To improve the early and late benefits (the patency and total myocardium revascularization) of coronary artery bypass grafting, stimulate us using only bilateral internal mammary artery (BIMA) composite Lima (left internal mammary artery)-Rima (right internal mammary artery) Y graft with off-pump, here is the early evaluation.</p><p><b>METHODS</b>From October 2002 to December 2005, 125 patients underwent off-pump coronary artery bypass grafting with the only composite grafts. The bilateral semi-skeletonization internal mammary artery pedicles composed the Y graft, the free Rima was anastomosed to the in situ Lima. The operation was performed off-pump and sequential anastomosis.</p><p><b>RESULTS</b>Four hundred and thirteen grafts for 125 patients, average number of grafts per patient was 3.3. Graft flow was measured with Transit time flowmeter during operation time. All grafts were patent during operation. There was no death perioperative period.</p><p><b>CONCLUSION</b>Off-pump coronary artery bypass grafting with only bilateral internal mammary artery composite Lima-Rima Y graft is secure and feasible. The technique could achieve total arterial myocardium revascularization and avoiding any procedure on the ascending aorta.</p>

FK228 blocks cells survival signal pathways and induces apoptosis of prostate cancer DU145 cells / 中国肿瘤生物治疗杂志

Objective:To investigate the underlying mechanism of histone deacetylase (HDAC) inhibitor FK228-in- duced apoptosis of the prostate cancer cell line DU145.Methods:The inhibitory effect of FK228 on DU145 cell growth and its cytotoxicity were determined by MTT assay;cell cycle arrest was detected by flow cytometry assay;morphological change was observed by Giemsa staining;and defined kinase protein levels were determined by Western blot analysis.Re- suits:FK228 obviously inhibited DU145 cells growth,arrested cell cycle at G_0/G_1 phase,induced cells morphological changes and degraded several kinase proteins,including EGFR,Her2,Raf-1,Src,Cdk4 and IAP member Survivin.The degradation of these kinases blocked Raf-Mek-Erk and PI3K/Akt survival signal pathways,inducing apoptosis.Condu- sion:FK228 may induce DU145 cell apoptosis through depletion of multiple kinase proteins and blockade of survival sig- nal pathways of DU145 cells.