LKB1 regulates epithelial-mesenchymal transition in Peutz-Jeghers hamartoma and intestinal epithelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the molecular mechanism by which LKB1 regulates epithelial-mesenchymal transition (EMT) in Peutz-Jeghers hamartoma and intestinal epithelial cells.</p><p><b>METHODS</b>Immunohistochemistry was used to detect gene expression of LKB1, E-cadherin, and vimentin in 20 hamartoma tissues and 10 normal intestinal tissues, and collagen fiber deposition was analyzed using Masson trichrome staining. Normal intestinal epithelial NCM460 cells were transfected with LKB1 shRNA plasmid or negative control via lentiviral vectors, and the role of LKB1 in cell polarization and migration were determined using CCK8 and Transwell assays. Western blotting, quantitative real-time PCR (qPCR) and immunofluorescence were used to assess the alterations of EMT markers in the cells with LKB1 knockdown.</p><p><b>RESULTS</b>Compared with normal intestinal tissues, hamartoma polyps showed significantly decreased LKB1 and E-cadherin expressions and increased vimentin expression with increased collagen fiber deposition. The cells with LKB1 knockdown exhibited enhanced cell proliferation and migration activities (P<0.01). Western blot analysis, qPCR and immunofluorescence all detected decreased E-cadherin and increased N-cadherin, vimentin, Snail, and Slug expressions in the cells with LKB1 knockdown.</p><p><b>CONCLUSION</b>s LKB1 deficiency triggers EMT in intestinal epithelial cells and Peutz-Jeghers hamartoma, suggesting that EMT can serve as the therapeutic target for treatment of Peutz-Jeghers syndrome.</p>

Effect of G-CSF and TPO on HIBD in neonatal rats

OBJECTIVE@#To observe effect of granulocyte colony-stimulating factor (G-CSF) and restructure human thrombopoietin on hypoxic-ischemic brain damage (HIBD) in new born rats.@*METHODS@#A total of 60 neonatal SD rats were selected and divided into 4 groups, with 15 in each group. Group A served as control group. Rats of Groups B-D were prepared for HIBD model by ligation of left common carotid artery combined with hypoxia method. Rats of Group A were only completed with free left common carotid artery without ligation and hypoxia operation. After HIBD model preparation, Group B was administrated with subcutaneous injection of normal saline for placebo treatment; Group C was administrated with cervical subcutaneous injection of 0.5 μg/10 g granulocyte colony stimulating factor (G-CSF) for 5 d (Once a day); Group D was administrated with intraperitoneal injection of 15 U/10 g recombinant human thromobopoietin (rhTPO) for treatment. After modeling for 7, 14 and 21 d, 5 rats were sacrificed in each group, respectively. Brain quality damage (%) conditions of experimental animals in each group were compared in different time points, and cerebral histopathological changes of each group were observed. Expression of nestin in rats of each group was detected by immunohistochemical method.@*RESULTS@#After modeling for 7, 14 and 21 d, brain quality damages (%) of Groups B, C and D were significant higher than that of in Group A (P0.05).@*CONCLUSIONS@#Both G-CSF and TPO can protect the nervous system of HIBD neonatal rats. G-CSF can promote the proliferation and differentiation of neural precursor cells to decrease the degeneration and necrosis of nerve cell. TPO can obviously ameliorate morphology index of HIBD rats. Through regulating ratio of TIMP-1 and MMP-9, TPO can maintain the integrity of blood brain barrier to relieve the occurrence of brain damage.

Effect of G-CSF and TPO on HIBD in neonatal rats

Objective: To observe effect of granulocyte colony-stimulating factor (G-CSF) and restructure human thrombopoietin on hypoxic-ischemic brain damage (HIBD) in new born rats. Methods: A total of 60 neonatal SD rats were selected and divided into 4 groups, with 15 in each group. Group A served as control group. Rats of Groups B-D were prepared for HIBD model by ligation of left common carotid artery combined with hypoxia method. Rats of Group A were only completed with free left common carotid artery without ligation and hypoxia operation. After HIBD model preparation, Group B was administrated with subcutaneous injection of normal saline for placebo treatment; Group C was administrated with cervical subcutaneous injection of 0.5 μg/10 g granulocyte colony stimulating factor (G-CSF) for 5 d (Once a day); Group D was administrated with intraperitoneal injection of 15 U/10 g recombinant human thromobopoietin (rhTPO) for treatment. After modeling for 7, 14 and 21 d, 5 rats were sacrificed in each group, respectively. Brain quality damage (%) conditions of experimental animals in each group were compared in different time points, and cerebral histopathological changes of each group were observed. Expression of nestin in rats of each group was detected by immunohistochemical method. Results: After modeling for 7, 14 and 21 d, brain quality damages (%) of Groups B, C and D were significant higher than that of in Group A (. P0.05). Conclusions: Both G-CSF and TPO can protect the nervous system of HIBD neonatal rats. G-CSF can promote the proliferation and differentiation of neural precursor cells to decrease the degeneration and necrosis of nerve cell. TPO can obviously ameliorate morphology index of HIBD rats. Through regulating ratio of TIMP-1 and MMP-9, TPO can maintain the integrity of blood brain barrier to relieve the occurrence of brain damage.

Nerve protective effect of Baicalin on newborn HIBD rats

OBJECTIVES@#To investigate the nerve protective effect and mechanism of baicalin on newborn rats with hypoxic ischemic brain damage (HIBD).@*METHODS@#A total of 64 SD newborn rats were randomly divided into control group, model group, nerve growth factor group and baicalin group, with 16 in each group. Left carotid artery ligation method was adopted to establish the HIBD model except for in control group, which was treated with intraperitoneal injection of salin e10 mL/kg for 3 d. After oxygen recovery on hypoxia ischemia rats, intraperitoneal injection of saline 10 mL/kg was adopted in model group for 3 d. Intraperitoneal injection of nerve growth factor injection 50 μg/kg per day was adopted in nerve growth factor group for 3 d; intraperitoneal injection of radix scutellariae 16 mg/kg per day was adopted in baicalin group for 3 d after modeling. Four rats of each group were sacrificed at Day 1, 2, 3, 7 for microscopic observation of pathological morphological changes in brain tissue after HE staining, S-P immunohistochemical method was used for observation of Fas and FasL expression in brain cells.@*RESULTS@#Neat structure of cells was observed in control group; edema cells in disordered arrangement was observed in model group, with some cells necrosis and cavity change; tissue injury in nerve growth factor group and baicalin group was significantly lighter than that in model group; Fas and FasL expression in model group, nerve growth factor group and baicalin group were significantly higher than that in control group at different time points (P0.05).@*CONCLUSIONS@#Baicalin can reduce expression of Fas and FasL in HIBD rats, inhibit apoptosis of nerve cells, thus achieve the protective effect on HIBD rat nerves.

Effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rats

OBJECTIVE@#To observe effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rats.@*METHODS@#A total of 75 SD rats were randomly divided into A, B, C, D, E groups with 15 in each group. Rats in group A served as the control group received just only but tissue separation without modeling operation, while model of unilateral ureteral obstruction (UUO) was established in B, C, D, E groups. Rats in A, B group were given saline lavage placebo treatment, while rats in C, D, E groups were given diammonium glycyrrhizinate and alprostadil injection. Five rats were sacrificed 1, 2, 3 weeks after modeling, serum creatinine level of femoral venous blood was determined. Transforming growth factor - β1 (TGF - β1) and concentration of connective tissue growth factor (CTGF) were also detected by using ELISA. Line renal interstitial tissue was taken after HE staining, renal interstitial TGF - β1 and CTGF expression were detected by using immunohistochemical method.@*RESULTS@#Serum creatinine levels of B, C, D, E group at different time points in were significantly higher than that of group A (P0.05); while serum and kidney tissue TGF - β1, concentration of CREA, expression of rats in B, C, D, E groups showed a gradual increasing trend over time. TGF - β1 and CREF of Group B in serum and kidney tissues at each time point were significantly higher than that of the other groups (P<0.05). TGF - β1 and CREF of Group E in serum and kidney tissues at each time point were significantly lower than that of B, C, D group at all time points in serum and kidney tissues (P<0.05).@*CONCLUSIONS@#Alprostadil combined with diammonium glycyrrhizinate can significantly lower the expression of TGF - β1 and CTGF in serum and tissues of SD rat with renal interstitial fibrosis, thus inhibit rat renal interstitial fibrosis process. It has synergy protective effect.

Effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rats

Objective: To observe effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rats. Methods: A total of 75 SD rats were randomly divided into A, B, C, D, E groups with 15 in each group. Rats in group A served as the control group received just only but tissue separation without modeling operation, while model of unilateral ureteral obstruction (UUO) was established in B, C, D, E groups. Rats in A, B group were given saline lavage placebo treatment, while rats in C, D, E groups were given diammonium glycyrrhizinate and alprostadil injection. Five rats were sacrificed 1, 2, 3 weeks after modeling, serum creatinine level of femoral venous blood was determined. Transforming growth factor - β1 (TGF - β1) and concentration of connective tissue growth factor (CTGF) were also detected by using ELISA. Line renal interstitial tissue was taken after HE staining, renal interstitial TGF - β1 and CTGF expression were detected by using immunohistochemical method. Results: Serum creatinine levels of B, C, D, E group at different time points in were significantly higher than that of group A (. P0.05); while serum and kidney tissue TGF - β1, concentration of CREA, expression of rats in B, C, D, E groups showed a gradual increasing trend over time. TGF - β1 and CREF of Group B in serum and kidney tissues at each time point were significantly higher than that of the other groups (. P<0.05). TGF - β1 and CREF of Group E in serum and kidney tissues at each time point were significantly lower than that of B, C, D group at all time points in serum and kidney tissues (. P<0.05). Conclusions: Alprostadil combined with diammonium glycyrrhizinate can significantly lower the expression of TGF - β1 and CTGF in serum and tissues of SD rat with renal interstitial fibrosis, thus inhibit rat renal interstitial fibrosis process. It has synergy protective effect.

Severe sepsis as an initial presentation in children with Wernicke' s encephalopathy: report of a case and literature review / 中华儿科杂志

<p><b>OBJECTIVE</b>Wernicke's encephalopathy (WE) is an acute neuropsychiatric syndrome resulting from thiamine deficiency, which is associated with significant morbidity and mortality. The disorder is still greatly underdiagnosed in children because of either a relatively non-specific clinical presentation in some cases or unrecognized clinical setting. The aim of this literature review was to provide knowledge of pediatric WE in an effort to assist in early diagnosis, thereby reducing the morbidity and mortality.</p><p><b>METHODS</b>The clinical manifestations, characteristic magnetic resonance imaging (MRI), diagnosis and treatment of one case and the other 35 cases reported in the last decade in children were summarized.</p><p><b>RESULTS</b>Thirty-six cases (22 boys and 14 girls, 2-month to 16-year-old) were analyzed. All the other 35 cases except for our case had underlying diseases: improper feeding in 25/35 cases, long-time vomiting in 5/35 cases, immunosuppressive therapy in 4/35 cases, long-time total parenteral nutrition without multivitamin preparations supplementation in 3/35 cases and anorexia nervosa in 1/35 case. The classic triad (mental-status changes, nystagmus and ophthalmoplegia, and ataxia) was seen in 6/36 cases. The other clinical manifestations included consciousness disturbance in 24/36 cases, infection in 22/36 cases, pathological reflex and muscular tension changes in 18/36 cases, convulsion in 17/36 cases, developmental delay in 4/36 cases and failure to thrive in 2/36 cases. Cerebrospinal fluid examination was performed in 31/36 cases, and a slightly raised protein concentration was seen in 7/31 cases. The cerebrospinal fluid lactate levels were detected in 4/36 cases (all increased), serum lactic acid levels in 7/36 cases (6/7 cases increased), serum pyruvate in 4/36 cases (all increased), thiamine pyrophosphate effect (TPPE) in 9/36 cases (all increased), and serum thiamine in 2/36 cases (increased in 1/2 cases). The brain computed tomography (CT) scan was conducted in 20/36 cases and 16/20 cases showed abnormal hypodensity in bilateral basal ganglia, one case revealed diffuse cortical atrophy. The brain MR scan was conducted in 13/36 cases and all the 13 cases revealed symmetrical abnormal signal in bilateral mamillary body and basal ganglia, and 7/13 cases showed abnormal signals in the tegmentum of midbrain, cerebral aqueduct and white matter around the third and fourth ventricles. The diagnosis of WE was confirmed by MR in 12 cases, triad combined with MR in 3 cases, autopsy in 1 case among the 13 cases who underwent MR scan. The diagnosis of WE was confirmed by the TPPE and/or lactate levels in 9/11 cases. The initial thiamine was given by intravenous or intramuscular infusion in 33/36 cases, unknown method in 1 case, orally in 1 case and no thiamine was used in 1 case. The dosage of thiamine was 100 mg daily in 29/35 cases, unknown in 3/35 cases, 50 mg daily in 2/35 cases, 600 mg daily in 1/35 case. 34/35 patients' clinical symptoms improved during 24 hours to 1 week after initial treatment, and 1 case died due to no response to thiamine. Nineteen patients were followed up for 2-2.5 months and 17 cases recovered completely.</p><p><b>CONCLUSION</b>Wernicke's encephalopathy can be difficult to diagnose because of a relatively non-specific clinical presentation. The characteristic MRI findings and the dramatic response of neurological signs to parenteral thiamine will assist early clinical diagnosis. Early and timely thiamine supplementation could reverse the clinical features and improve the prognosis in most cases.</p>

Effects of TrkB-BDNF signal pathway on synthesis and secretion of vascular endothelial growth factor in human neuroblastoma cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of TrkB-BDNF signal pathway on the synthesis and secretion of vascular endothelial growth factor (VEGF) in human neuroblastoma cells (NB).</p><p><b>METHODS</b>TrkB protein expression in SY5Y cells before and after all-trans-retinoicacid (ATRA) treatment was detected by Western blot. P-TrkB protein expression in SY5Y cells before and after the treatment of ATRA along with BDNF was also detected by Western blot. VEGF concentrations in the SY5Y cell culture supernatants were measured using ELISA after the treatment with ATRA, BDNF, tyrosine kinase inhibitor K252a and PI3k inhibitor LY294002.</p><p><b>RESULTS</b>TrkB protein was undetectable in SY5Y cells before ATRA treatment. After the treatment of 1, 10 and 100 nM/L ATRA for five days, TrkB protein was expressed in SY5Y cells and the TrkB protein level increased with the increasing ATRA concentration. P-TrkB protein was not expressed in SY5Y cells treated only with 10 nM/L ATRA, but it was detectable after the treatment of ATRA along with BDNF. VEGF concentrations in the group treated with ATRA+BDNF were significantly higher than those in the untreated control and the ATRA alone treatment groups (P<0.01). VEGF concentrations in the K252a pretreated ATRA+BDNF group were significantly lower than those in the group treated with ATRA+BDNF (P<0.05). VEGF concentrations in the LY294002 treatment group (ATRA+LY294002+BDNF group) were also significantly lower than those in the group treated with ATRA+BDNF (P<0.01).</p><p><b>CONCLUSIONS</b>Activation of TrkB-BDNF signal pathway may increase the synthesis and secretion of VEGF in human NB cells. The synthesis and secretion of VEGF can be inhibited by blocking TrkB-BDNF signal pathway with K252a or blocking the TrkB-BDNF downstream signal pathway PI3K/Akt with LY294002.</p>

Statistical study on correlation between cerebral arteriovenous malformation and hemodynamic aneurysms / 中华外科杂志

<p><b>OBJECTIVE</b>to explore the characteristic factors of arteriovenous malformation (AVM) which have statistically significant correlation with hemodynamic aneurysms.</p><p><b>METHODS</b>from August 1999 to July 2009, the clinical and imaging indices of 363 consecutive patients with AVM were retrospectively reviewed and entirely statistically analyzed. There were 229 male patients and 137 female patients, the mean age at the time of presentation was 28 ± 13 years. By using SPSS 16.0 medical statistic software, the correlation were analyzed between hemodynamic aneurysms and 13 characteristic factors associated with AVM through the methods of unit-factor and multi-factor analysis. Finally, the risk of the correlative factors filtered were evaluated.</p><p><b>RESULTS</b>the crosstabs analysis of unit-factor strongly suggested that the following factors, including age, location (supertentorium, subtentorium), size, number of main feeding arteries, number of drainage veins, ectasis of drainage veins, contralateral supply, and supply by both anterior and posterior circulation, were correlated with hemodynamic aneurysms. And the results of regression analysis of multi-factors indicated the following factors, including age, number of main feeding arteries, and contralateral supply, were positively correlated with hemodynamic aneurysms and the number of drainage veins were negatively correlated with hemodynamic aneurysms.</p><p><b>CONCLUSION</b>the factors including age, number of main feeding arteries, number of drainage veins and contralateral supply, are highly correlated with hemodynamic aneurysms.</p>

Observation of special staining of human colorectal carcinoma cell lines cultured on nitrocellulose membrane / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the special staining of cells cultured on nitrocellulose (NC) membrane and evaluate the application of the novel method for cell culture and pathological staining.</p><p><b>METHODS</b>Human colorectal carcinoma SW1116 cell line and SW480 cell line were cultured using nitrocellulose membrane as the culture matrix, with the same cells cultured on slides serving as the control.</p><p><b>RESULTS</b>The cells cultured on NC membrane appeared transparent with sharp edge and purple background by macroscopic observation, showing on obvious difference in terms of cell morphology and number from the cells cultured on glass slides. Irregular polygonal SW1116 cells and SW480 cells were found on the NC membrane, on which the cells grew in colony and showed blue nucleus and red cytoplasm.</p><p><b>CONCLUSIONS</b>NC membrane produces no cytotoxicity and can be used for cell culture without affecting the normal cell morphology and number during cell culture, thus providing a new means for cell culture and pathological staining.</p>

Effects of therapeutic hypercapnia on inflammation and apoptosis after hepatic ischemia-reperfusion injury in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Therapeutic hypercapnia (TH) has been demonstrated to protect several organs ischemia-reperfusion injury. The study aimed to investigate the effects of therapeutic hypercapnia on hepatic ischemia-reperfusion injury (HIRI).</p><p><b>METHODS</b>Thirty adult male Wistar rats weighing (250+/-20) g were randomized into 3 groups (n=10 in each), group C (control group), group A (hypercapnia group) and group B (CO2 preconditioning group). A segmental ischemia of the liver was induced by interrupting the blood vessels including the bile duct to the median and left lateral lobes for 60 minutes and all the animals were sacrificed after 240 minutes observation period of reperfusion. Mean arterial pressure (MAP) and the blood gases were measured before ischemia (baseline) and at 30, 60, 120, 180 and 240 minutes after reperfusion. Arterial blood samples were obtained for determination of serum levels of TNF-alpha, IL-10, serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The histopathology of liver tissues was evaluated by light microscopy. The NF-kappaB expression and apoptotic hepatocytes were respectively determined by immunohistochemistry and TUNEL assay.</p><p><b>RESULTS</b>The serum levels of liver enzymes and TNF-alpha were significantly decreased while the IL-10 level was significantly increased in groups A and B than in group C (P<0.05), and group B surpassed group A (P<0.05). The histopathological scores, the NF-kappaB immunohistochemical score (IHS) and apoptotic index were significantly lower in groups A and B than in group C (P<0.05), and the decrease in group B was more obvious than in group A (P<0.05).</p><p><b>CONCLUSION</b>Therapeutic hypercapnia attenuates ischemia-reperfusion injury to the liver. Moreover, the effects of CO2 preconditioning are outstandingly notable.</p>

The relationship between uncoupling protein 2 expression and myocardial high energy phosphates content in abdominal aorta constriction induced heart failure rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the changes of expression of uncoupling protein 2 (UCP2) in pressure overload induced failure myocardium in rats.</p><p><b>METHODS</b>Male SD rats were randomized into 3 groups (n = 15 each): abdominal aorta constriction (AC) 20 weeks group (H20w group), sham operation group (SH20w group) and normal control group (N group). Twenty weeks later, myocardial function was evaluated by echocardiography and hemodynamic measurements. Mitochondria in ventricular tissue were isolated by centrifugation. Adenine nucleotide pools (ATP, ADP, AMP, PCr) in myocardium were measured by high performance liquid chromatography. The expression of UCP2 in mitochondria was detected by PT-PCR and Western blot analysis.</p><p><b>RESULTS</b>Myocardial function was significantly decreased 20 weeks post-AC compared to SH20w group and N group. Myocardial ATP, ADP, AMP and PCr contents were also significantly decreased in H20w group than the other 2 control groups. The expression of UCP2 in myocardial mitochondria was significantly increased in H20w group and negatively correlated with ATP contents (r = -0.929, P < 0.01).</p><p><b>CONCLUSIONS</b>The expression of UCP2 was upregulated in pressure overload induced failure heart and might be responsible for decreased myocardial adenine nucleotide and energy metabolism disturbance in this model.</p>

Effect of renshen jianxin capsule for alleviating insulin resistance in patients with coronary heart disease and glucose tolerance impairment / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of Renshen Jianxin Capsule (RJC) on insulin resistance in patients with coronary heart disease (CHD) and glucose tolerance impairment (GTI).</p><p><b>METHODS</b>Eighty patients with CHD-GTI of qi-deficiency blood-stasis syndrome were randomly assigned to 2 groups equally, the treated group was treated by RJC and the control group by metformin, based on the conventional Western medical treatment with nitric esters for 20 weeks. Changes before and after treatment in clinical symptoms and levels of blood glucose insulin, and insulin sensitivity index (ISI) were observed.</p><p><b>RESULTS</b>The scores of clinical symptoms of Chinese medicine decreased in both groups, which showed statistical significances compared with those before treatment (P<0.01, P<0.05). On alleviating the angina pectoris, the markedly effective rate of the treated group is 47.5%, the total effective rate was 80.0%, and the difference between the two groups showed statistical significance (P<0.05). FBG, INS and ISI were improved significantly in both groups after treatment (P<0.05, P<0.01); while the three indices showed in significant difference between the two groups after treatment (P> 0.05).</p><p><b>CONCLUSION</b>RJC was effective in improving insulin resistant, which may be one of the mechanisms of its therapeutic effect on CHD.</p>

Sorafenib reverses multidrug resistance of hepatoma cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the role of sorafenib in reversing multidrug resistance (MDR) in hepatoma BEL-7402/FU cells and its possible mechanisms.</p><p><b>METHODS</b>MTT colorimetric assay was used to obtain the dose-response curve of sorafenib in BEL-7402/FU cells, and flow cytometry performed to assess the effect of sorafenib on Rho123 concentration in the cells. The optimal dose of sorafenib for cell treatment was determined according to the results of MTT assay and flow cytometry. MTT assay was employed to evaluate the effect of sorfenib on the cytotoxicity of the antitumor drugs, flow cytometry performed to determine the expression of cell membrane transport protein (P-gp), and RT-PCR used to detect mdr1 gene expression in the cells treated with sorafenib at the optimal dose.</p><p><b>RESULTS</b>Sorafenib at the concentration of 4 micromol/L, efficiently reversed the MDR of the cells with minimal side effects. At the concentration of 4 micromol/L, sorafenib partially reversed the drug resistance of BEL-7402/FU cells to ADM, 5-FU, GEM and DDP, with reversal indexes of 2.98, 7.16, 1.99 and 10.08, respectively. Treatment of the cells with 4 micromol/L, sorafenib also partially down-regulated P-gp expression in BEL-7402/FU cells, and caused a reduction of mdr1 gene expression by 27.3% in comparison with the control cells.</p><p><b>CONCLUSION</b>Sorafenib can reverse MDR in human hepatoma cells probably in association with down-regulation of mdr1 gene expression and increased accumulation of the chemotherapeutic agents in the cells.</p>

Association among circulating endothelial progenitor cells, insulin resistance and severity of coronary lesions in patients with coronary artery disease / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the correlation between the number and activity of circulating endothelial progenitor cells (EPCs), insulin resistance and severity of coronary lesions in patients with coronary artery disease (CAD).</p><p><b>METHODS</b>Patients with coronary angiography evidenced CAD were divided in insulin resistance group (IR, n = 25) and insulin sensitive group (IS, n = 44) according to insulin level, 25 health volunteers served as control. Circulating EPCs were marked as KDR/CD133+ cells via fluorescence-activated cell sorter analysis. EPCs were also isolated from peripheral blood and cultured in vitro for 7 days, identified by DiI-acLDL uptake and lectin staining methods. EPCs migration activities were determined by modified Boyden chamber assay, EPCs proliferation activities were determined by MTT assay.</p><p><b>RESULT</b>Circulating EPCs number was significantly lower in IR group compared with IS group [(0.34 +/- 0.08) per thousand vs. (0.47 +/- 0.09) per thousand, P < 0.01] and control group (P < 0.05). Both insulin resistance index (r = -0.291, P = 0.01)and Gensini score (r = -0.3984, P = 0.006)were negatively correlated with number of circulating EPCs. Proliferation and migration capacities of EPCs were also significantly lower in IR group compared to those in IS group (all P < 0.05) and control group (all P < 0.05).</p><p><b>CONCLUSIONS</b>Insulin resistance/hyperinsulinemia could aggravate severity of coronary artery lesions via reducing the number and activities of circulating EPCs in patients with CAD.</p>

Blocking TrkB-BDNF signal pathway decreases the livability of neuroblastoma cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Brain-derived neurotrophic factor (BDNF) and its specific tryrosin kinase receptor-B (TrkB) are highly correlated to the chemoresistance of neuroblastoma (NB) cells and poor prognosis. This study observed the changes of the sensibility of NB cells to chemotherapy drug cisplatin (CDDP) before and after blockage of TrkB-BDNF signal pathway by specific tyrosin kinase inhibitor K252a.</p><p><b>METHODS</b>Human NB cell line SH-SY5Y (SY5Y) was routinely cultured. Expression of TrkB was induced with nM all trans-retinoid acid (ATRA). Then BDNF, CDDP or K252a were added to the cultured SY5Y cells. Cell livability was assessed by methyl thiazolyl tetrazolium (MTT) assay. TrkB autophosphorylation was determined by Western blot analysis. Cell apoptosis rate was detected by flow cytometry (FCM). The conformation of apoptosis cells was observed by transmission electron microscopy (TEM).</p><p><b>RESULTS</b>The livability and apoptosis rate in SY5Y cells treated with ATRA, BDNF and CDDP were not different from the blank control group. However, after K252a together with ATRA, BDNF and CDDP treatment, the sensibility of SY5Y cells to chemotherapy drug CDDP increased, the livability decreased and the apoptosis rate increased in SY5Y cells when compared with the blank control group (P <0.01). K252a treatment resulted in blockage of TrkB autophosphorylation.</p><p><b>CONCLUSIONS</b>The blockage of TrkB-BDNF signal pathway by K252a use can increase sensibility of NB cells to chemotherapy and thus decrease the livability of NB cells.</p>

Morphological changes of blood spleen barrier in portal hypertensive spleen / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The pathogenesis of hypersplenism and the immune function of the spleen in patients with portal hypertension (PH) remain obscure. This study aimed to evaluate the morphological changes of blood spleen barrier in spleen with hypersplenism due to PH and provide evidence for an in-depth investigation of the immune function of the spleen with hypersplenism and the mechanism of hypersplenism.</p><p><b>METHODS</b>Spleen samples from 12 portal hypertensive patients and 4 patients with traumatic ruptures of spleen were examined. The samples of spleen were made into pathological sections, stained with Masson trichrome stain, Gomori stain, and CD68, CD34 immunohistochemistry, and were examined microscopically for the changes in the distribution of collagen fibers, reticular fibers, macrophages, and vascular endothelial cells. The changes in ultrastructure of macrophages and endothelial cells in marginal zone were also evaluated by transmission electron microscopy.</p><p><b>RESULTS</b>As compared to the normal spleen, the density of macrophage in the PH spleen was decreased, but the macrophages were mainly located in the marginal zone and distributed around the splenic corpuscle, with many villi and pseudopodium-like protrusion on the cell surface. The accrementition of collagen fibers was obvious around the splenic corpuscle and central artery. The increased reticulate fibers encircled the splenic corpuscle with more connection between the fibers. The vascular endothelial cells were in diffused distribution, without any regionality in PH spleen, but the vessel with enlarged lumina increased in red pulp.</p><p><b>CONCLUSIONS</b>The morphological changes of the blood spleen barrier can be one of the pathological fundaments for the abnormality of the immune function and the increased destruction of blood cells located in the spleens of patients with PH. However, this still entails clarification.</p>

Radioimmunotherapy with 131 I-labeled humanized anti-HBsAg Fab and anti-nucleus antigen monoclonal antibody chTNT in nude mice bearing human hepatocellular carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the feasibility and efficacy of radioimmunotherapy with 131 I-labeled humanized anti-HBsAg Fab (131 I-anti-HBsAg Fab) combined with 131 I-labeled anti-nucleus antigen monoclonal antibody chTNT (131 I-chTNT) in nude mice bearing human hepatocellular carcinoma.</p><p><b>METHODS</b>Nude mice bearing subcutaneous human hepatocellular carcinoma xenografts were treated by intratumoral injection of 131 I-anti-HBsAg Fab and/or 131 I-chTNT, and the changes in the tumor size and alterations in the radioactivity concentration in the tumor and non-tumor tissues were observed.</p><p><b>RESULTS</b>The tumor inhibition rate in mice treated with 131 I-anti-HBsAg Fab combined with 131 I-chTNT (73.09%) was significantly higher than that in mice treated with 131 I-anti-HBsAg Fab (47.8%) or 131 I-chTNT (54.26%) alone. Combined treatment also resulted in significantly higher tumor-to-normal radioactivity concentration ratios than the treatment with the single agents.</p><p><b>CONCLUSION</b>Intratumoral injection with 131 I-labeled monoclonal antibodies can increase the radioactivity concentration in the tumor and enhance the efficacy of the radioimmunotherapy in nude mice bearing human hepatocellular carcinoma.</p>

Identification of human coronary artery endothelial cells obtained by coronary endovascular biopsy / 中华心血管病杂志

<p><b>OBJECTIVE</b>To develop a method to obtain and identify human coronary artery endothelial cells obtained during percutaneous coronary interventions (PCI).</p><p><b>METHODS</b>Coronary guide wires were used to obtain endothelial cells from coronary arteries in 28 patients undergoing PCI. The cells were eluted from the wire tips and then purified by magnetic beads coated with anti-CD146 antibody. von Willebrand factor (vWF) was used as an immunocytochemical marker for endothelial cells. The cellular viability was evaluated by observing cell membrane integrity and energy-dependent uptake of DiI-labeled acetylated low-density lipoprotein.</p><p><b>RESULTS</b>An average of 96 coronary artery endothelial cells with good viability per patient were obtained by one guide wire. vWF identification showed their endothelial morphology and immunoreactivity.</p><p><b>CONCLUSION</b>The viable coronary endothelial cells could be obtained during routine percutaneous coronary interventions combined with magnetic beads isolation technique. These cells may be used for further cellular functional analyses (such as immunocytochemistry and molecular biology) and expand our understanding on mechanisms of coronary artery diseases.</p>

Inhibitory effects of recombinant human endostatin on the metastatic potential and invasiveness of hepatocellular carcinoma HCCLM6 cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effects of recombinant human endostatin (rh-ES) on cell adhesion, metastatic potential and invasiveness of hepatocellular carcinoma HCCLM6 cells in vitro.</p><p><b>METHODS</b>The changes in the cell proliferation status of HCCLM6 cells treated with different concentrations of rh-Endostatin in vitro was measured with MTT assay, and their invasiveness and migration were assayed using transwell cell culture chamber. The cell adhesion assay was carried out on 96-well plate precoated with matrigel.</p><p><b>RESULTS</b>Rh-ES showed inhibitory effect against the proliferation of HCCLM6 cells after a 72-h treatment. The adhesion, metastatic potential and invasiveness of the cells were obviously inhibited by rh-Endostatin in a dose-dependent manner.</p><p><b>CONCLUSION</b>Rh-ES inhibits the adhesion, invasiveness and migration of hepatocellular carcinoma cells in vitro, and the mechanism needs further investigation.</p>