Effect of Astragalus Polysaccharide on Growth and Tumor-related Apoptosis Protein of Human Breast Cancer MDA-MB-231 Transplanted Tumor in Nude Mice / 中国实验方剂学杂志

Objective: To observe the inhibitory effect of astragalus polysaccharides (APS) on growth of human breast cancer MDA-MB-231 xenograft tumor in nude mice and its effect on the apoptosis of tumor cells, in order to study the effect of APS on growth and induction of apoptosis of triple negative breast cancer MDA-MB-231 and its possible molecular mechanism. Method: Human breast cancer cell MDA-MB-231 was inoculated into the right axillary subcutaneous of BALB/c-nu female nude mice to establish the transplanted tumor model of breast cancer. Eighteen nude mice were randomly divided into 3 groups:model group (saline per day), low-dose APS group (200 mg·kg-1 APS per day), and high-dose APS group (400 mg·kg-1 APS per day), with 6 rats in each group. The drug was administered by gavage (200 μL) daily for 21 days. In the experiment, the length and diameter of breast cancer transplanted tumor were measured every two days, and the tumor volume was recorded and calculated. At the end of the experiment, the changes of tumor mass and tumor volume of the low and high-dose APS groups and the model group were observed and compared, and the tumor inhibition rate was calculated. The cell morphology in tumor tissue was observed by hematoxylin-eosin (HE) staining, and Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) was used to verify the apoptosis of breast cancer tissues. The expressions of apoptosis-related proteins, such as B-cell lymphoma/leukemia-2 protein (Bcl-2), Bcl-2 associated X protein (Bax), Caspase in tumor tissues was detected by Western blot. Result: The tumor volume of breast cancer decreased in the low and high-dose APS groups, and the tumor inhibition rates were 37.9%and 57.57%, respectively, with statistically significant differences from the model group (PP0.01). HE of tumor tissue cells showed that APS led to obvious morphological changes, with apoptosis in the tissue cells. TUNEL staining showed that the apoptosis rate of tumor cells in APS intervention groups was higher than that in control group. Western blot showed that expression of Bcl-2 protein decreased(PPPPConclusion: APS can effectively inhibit the growth of MDA-MB-231 breast cancer xenografts in nude mice and induce apoptosis in human breast cancer MDA-MB-231 cells. The mechanism may be related to the effect of APS on expressions of apoptosis-related proteins Bcl-2, Bax, Caspase-9 and Caspase-7 in breast cancer cells.

Clinical manifestations and gene analysis of 2 Chinese children with cystic fibrosis / 中华儿科杂志

<p><b>OBJECTIVE</b>Cystic fibrosis (CF) is rare in Chinese mainland. We present two cases of CF patients diagnosed by gene analysis. Their clinical manifestations and genetic mutation features are analyzed in this article. It will be of special interest to pediatricians in recognition of CF.</p><p><b>METHOD</b>The clinical material of two CF patients who were diagnosed by gene analysis was retrospectively analyzed.</p><p><b>RESULT</b>The first patient is a 13-year-old girl. She had a complaint of recurrent fever and cough for 6 months, expectoration for 2 months and hemoptysis for 20 days. After 3 months of her birth, she was operated on for bullae of lung. She was susceptible to upper respiratory tract infection. There was no family history of recurrent wheeze and other special diseases. Aspergillus fumigatus specific IgE was at grade 3 and aspergillus fumigatus IgG was high. Pseudomonas aeruginosa was positive in sputum culture. Sweat testing was performed and Na+ was higher. Pulmonary CT indicated bronchiectasis. Nasal sinus CT showed optical density of soft tissue within maxillary sinus and chronic bilateral sinusitis. The electron microscopy of cilia suggested immobile cilia syndrome. A heterozygotic mutation (263T > G, 2909G > A) was found after CFTR genetic mutation analysis. Both her parents were carriers. She was treated with inhalation of nebulized hypertonic saline and postural drainage for a long time. And she got better during a follow up period of 1 year. The second patient was a 10-year-old girl who complained of recurrent expectoration for 3 years and shortness of breath for half a year. She had a history of sinusitis and steatorrhea. The family history was normal. Both the lipase and insulin level in blood serum was lower.Pseudomonas aerugino and Aspergillus fumigatus were both positive in sputum culture. Aspergillus fumigatus IgE was normal. Pulmonary CT indicated bronchiolitis and bronchiectasis. Nasal sinus CT showed bilateral maxillary sinusitis. CFTR genetic mutation analysis revealed a homozygous mutation (3196C > T). Her parents and relatives did not participate in this study. Unfortunately, this child died of respiratory failure 3 months after discharge.</p><p><b>CONCLUSION</b>CFTR gene mutation was a main cause of CF. Common symptoms are those of bronchiectasis, pancreatitis and sinusitis. The two Chinese patients were diagnosed by gene analysis. One had a heterozygous mutation (263T > G, 2909G > A) and the other had a homozygous mutation (3196C > T), not ΔF508 which is common in western countries.</p>

Clinical features and molecular analysis of 2 Chinese children with autosomal recessive chronic granulomatous disease caused by CYBA mutations / 中华儿科杂志

<p><b>OBJECTIVE</b>To summarize clinical and molecular features of two children with autosomal recessive chronic granulomatous disease caused by CYBA mutations.</p><p><b>METHOD</b>The clinical records and CYBA mutations were reviewed for analysis of infections and inflammatory complications.</p><p><b>RESULT</b>The first case was a girl diagnosed with "liver and spleen abscess" in our hospital when she was 2.9 years old, with past history of neonatal impetigo and recurrent purulent lymphadenitis and positive family history. The results of DHR123 flow-cytometry showed that positive phagocytes after phorbol ester (PMA) stimulation was 84.63%. CYBA mutation analysis showed that she had heterozygous 35C > T, Q3X and IVS-2A > G. The second case was a boy diagnosed with "sepsis (salmonella D)" when he was 4 years old with a past history of impetigo, sepsis, perianal abscess, skin infection and positive family history. The results of flow cytometry showed that positive phagocytes after PMA stimulation was 96.13%. CYBA mutation analysis showed that he had homozygous 35C > T, Q3X and his parents were all carriers. All of them had BCG related axillary lymphnode calcification.</p><p><b>CONCLUSION</b>A22CGD cases had recurrent purulent infections (skin, lymphnode, liver and spleen, lung, blood), DHR123 flow cytometric analysis helped the diagnosis of CGD, CYBA mutation analysis ascertained the diagnosis of A22CGD.</p>

Screening and application of prokaryotic enhancer-like sequence 3A / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To screen enhancer-like sequences from Escherichia coli strain C600 genome, to construct an expression vector harboring prokaryotic enhancer-like sequence and study the effect of interferon gene expression.</p><p><b>METHODS</b>Enhancer-like element from Escherichia coli strain C600 genome was obtained by using the chloramphenicol acetyl-transferase (CAT) gene as reporter gene. An expression vector harboring prokaryotic enhancer-like sequence from Escherichia coli strain C600 was constructed. Interferon was expressed and assayed.</p><p><b>RESULTS</b>An enhancer-like sequences with distance and orientation independence property were screened and named 3A. Quantification test showed that the direct and reverse orientation of 3A could increase the activity of beta-galactosidase with 7.11 and 2.93 times. The enhancing activity of the element was on transcription level. An expression vector harboring the prokaryotic enhancer-like sequence 3P3 which was enhancing function region of sequence 3A was constructed. Using this vector the antiviral activity of interferon alpha-2b was increased by 3.7 times in comparison with the original expression plasmid.</p><p><b>CONCLUSION</b>3A enhancer-like sequence was screened from Escherichia coli strain C600 genome. Interferon gene was highly expressed by using an expression vector harboring enhancer-like sequences.</p>

Clinical analysis of 18 children with disseminated Bacille Calmette-Guérin infection / 中华儿科杂志

<p><b>OBJECTIVE</b>To explore the clinical manifestation, immune abnormality and outcome of disseminated Bacille Calmette-Guérin (BCG) infection in children.</p><p><b>METHOD</b>The clinical data of 18 children with disseminated BCG infection seen from January 2000 to December 2007 were analyzed retrospectively.</p><p><b>RESULT</b>Thirteen of the children were male among 18 patients. Disseminated infection first appeared in armpit lymph nodes ipsilateral to the vaccination site, then spread to lungs in 15, lymphnodes of mediastinum or abdominal cavity in 18, skin and soft tissues in 5, skeletons in 4, liver in 4, spleen in 8, kidney, adrenal gland or meninges in 3. Twelve children were diagnosed to have primary immunodeficiency; 3 had severe combined immunodeficiency (SCID); 7 had chronic granulomatous disease (CGD), 2 had IL-12/IFN-gamma passageway deficiency. Eleven of the 18 patients died, and the remaining 7 patients were followed up from 1 to 9 years and are alive at present, but presented recurrent skin and bone tuberculosis in 4 and recurrent other infection in 3.</p><p><b>CONCLUSION</b>Most Children with disseminated BCG infection had primary immunodeficiency. CGD and IL-12/IFN-gamma passageway deficiency accounted for considerable proportion, so special immune function should be detected in these patients. The prognosis was poor. The type of the immunodeficiency diseases should be identified in early stage and the specific immune treatment should be given to the patients.</p>

Effects of oxygen and calcium on the expression of wild-type and mutated hypoxia-inducible factor-1alpha eukaryotic vectors in HEK293 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effects of oxygen and calcium on the expression of eukaryotic vectors harboring wild-type or mutated hypoxia-inducible factor-1alpha (HIF-11alpha) in HEK293 cells.</p><p><b>METHODS</b>HEK293 cells were transiently transfected with pcDNA3.1+/HIF-11alpha, pcDNA3.1+/HIF-11alpha-564Ala and pcDNA3.1+/HIF-11alpha-564Ala-803Ala via lipofectin. Western blotting were used to detect HIF-11alpha protein after normoxic or hypoxic exposure of the transfected HEK293 cells in the presence or absence of Ca(2+). The levels of vascular endothelial growth factor (VEGF) mRNA in the transfected cells in normoxic condition was detected using RT-PCR.</p><p><b>RESULTS</b>The levels of HIF-11alpha protein and VEGF mRNA increased in HEK293 cells transfected with the vectors harboring mutated HIF-11alpha, but not in the cells transfected with wild-type HIF-11alpha vectors in normoxia. Hypoxia increased the levels of HIF-11alpha protein in the cells transfected with wild-type HIF-11alpha vectors, which was inhibited by the application of Ca(2+). Ca(2+) showed no inhibitory effect on HIF-11alpha levels in HEK293 cells transfected with the vectors containing mutated HIF-11alpha.</p><p><b>CONCLUSION</b>The protein products of pcDNA3.1+/HIF-11alpha-564Ala and pcDNA3.1+/HIF-11alpha- 564Ala-803Ala in HEK293 cells enhance the cell tolerance to oxygen and protease.</p>

Expression of B7-related protein-1 on human coronary artery endothelial cells and its clinical significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of oxidized low density lipoprotein (ox-LDL) on the expression of B7-related protein-1 (B7RP-1) on human coronary artery endothelial cells (HCAECs).</p><p><b>METHODS</b>HCAECs were incubated in the presence of 100 mg/L ox-LDL for 24 h, and B7RP-1 expression levels were determined using fluorescence reverse transcription PCR (RT-PCR) and Western blotting.</p><p><b>RESULTS</b>B7RP-1 expression was detected HCAECs, with spots of fluorescence signals distributing on the cell membrane as observed under fluorescence microscope. RT-PCR with B7RP-1 specific primers yielded products of expected size (496 bp). Western blotting identified B7RP-1 expression in the HCAECs as a cell-associated protein with an apparent molecular mass of 70,000. Treatment of the cells with ox-LDL significantly increased B7RP-1 expression at both the mRNA and protein levels (P<0.05).</p><p><b>CONCLUSION</b>B7RP-1 is expressed on the membrane of HCAECs. ox-LDL can promote up-regulate the expression of B7RP-1, which might be one of the immunopathogenesis of atherosclerosis.</p>

Effect of porcine relaxin on NO production of human microvascular endothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe effect of porcine relaxin(pRLX) on NO production of human microvascular endothelial cells(HMVECs) and discuss its possible mechanism.</p><p><b>METHODS</b>iNOS and cNOS expression of HMVECs with or without pRLX were detected using western blotting. NO production of HMVECs with pRLX at different concentration or different time were determined by method of Griess. NO production of pRLX of HMVECs plus Non-selective NOS inhibitor NG-monomethyl-L-arginine(L-NMMA), selective iNOS inhibitor aminoguanidine(AG) or nuclear factors-kappaB (NF-kappaB) inhibitor pyrrolidine dithiocarbamate(PDTC) were also analysed.</p><p><b>RESULTS</b>pRLX promoted iNOS protein expression of HMVECs, but not cNOS protein expression. NO production of HMVECs was promoted by pRLX on concentration-dependent pattern instead of time-dependent one. AG, L-NMMA and PDTC were showed to block the effect of pRLX on NO production of HMVECs.</p><p><b>CONCLUSION</b>pRLX promote iNOS expression and NO production of HMVECs.</p>

Clinical features of 17 cases of X-linked agammaglobulinemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate clinical features of X-linked agammaglobulinemia (XLA) in children.</p><p><b>METHODS</b>The medical records of 17 children with XLA between January 2001 and April 2007 were reviewed.</p><p><b>RESULTS</b>The age at first diagnosis in 88.2% of patients was more than 6 years, with a mean of 7.7 years. Twelve patients (70.6%) presented first symptoms over 2 years old, with a mean of 4.2 years. Respiratory infections as first symptoms and complaints occurred in 64.7% of the patients and 35.3% of the patients presented with polyarthritis. Skin and soft tissue infections were rarely seen in less than 1 year old group children. Abrupt sepsis and abscess in deep tissues were seen in the older children. CD4+ T cells decreased and CD8+ T cells increased in 9 patients and an inversed ratio of CD4+/ CD8+ was observed in 11 patients.</p><p><b>CONCLUSIONS</b>Both the age presenting first symptoms and the age at first diagnosis in children with XLA in this study were later than the reported data. Respiratory infection was the most common manifestation. High prevalence of polyarthritis was observed. Abnormal T cell phenotypes occurred in more than one half of patients.</p>

Clinical features of coronary artery spasm patients with or without myocardial bridge / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the clinical features of coronary artery spasm patients with or without myocardial bridge and explore the roles of endothelial dysfunction in these patients.</p><p><b>METHODS</b>One hundred eighteen patients undergone acetylcholine provoking test were divided into myocardial bridge (MB) group (n = 26) and non-myocardial bridge (NMB) group (n = 92). The results of acetylcholine test, treadmill exercise electrocardiography, stress myocardial perfusion scintigraphy, plasma level of endothelin-1 and nitric oxide were compared between MB group and NMB group.</p><p><b>RESULTS</b>Coronary artery spasm was induced in 21 patients in MB group (81%) and 52 patients in NMB group (57%, P < 0.05). Positive treadmill electrocardiography was obtained in 19 patients in MB group (73%) and 7 patients in NMB group (8%, P < 0.001). Ischemic perfusion defect in 20 (77%) and 9 patients (10%, P < 0.001) and reverse redistribution in 23 (88%) and 68 patients (74%, P > 0.05). Patients showed different clinical features in MB group and NMB group (more short-duration exertional angina and could not be readily released by nitroglycerine in MB group while more patients experienced long-lasting variant angina and symptoms could be readily released by nitroglycerine). Plasma endothelin-1 level was significantly higher [(132.1 +/- 6.5) ng/L vs. (108.5 +/- 8.2) ng/L, P < 0.01] while nitric oxide was significant lower [(84.7 +/- 17.5) ng/L vs. (99.8 +/- 18.2) ng/L, P < 0.05] in MB group compared to NMB group.</p><p><b>CONCLUSION</b>MB patients were prone to coronary artery spasm partly due to endothelial dysfunction. Patients with MB and coronary artery spasm also showed classic clinical symptoms and positive stress tests for ischemia.</p>

Comparison of plasma low-density lipoprotein and oxidized low-density lipoprotein levels with coronary lesion severities in patients with coronary artery disease / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the relationship between plasma low-density lipoprotein (LDL) and oxidized low-density lipoprotein (ox-LDL) levels and the severity of coronary atherosclerosis.</p><p><b>METHODS</b>Fasting plasma ox-LDL was measured by enzyme-linked immunosorbent assay and plasma LDL was measured by biochemical autoanalyser in 31 patients with coronary artery spasm (CAS group, chest pain with positive acetylcholine provocation test but without significant coronary artery stenosis), 35 patients with stable angina pectoris (SAP group) and 24 healthy persons (control group).</p><p><b>RESULTS</b>Plasma LDL levels were similar between CAS and SAP groups but significantly higher than that in control group. Plasma ox-LDL levels significantly increased in proportion to coronary lesion severities [SAP (575 +/- 219 microg/L) > CAS (299 +/- 117 microg/L) > control (218 +/- 35 microg/L)]. In SAP group, plasma ox-LDL level was also significantly higher in multi-vessel disease group than that in single-vessel disease group (672 +/- 92 vs. 462 +/- 72 microg/L, P < 0.05).</p><p><b>CONCLUSION</b>Plasma ox-LDL but not LDL level is significantly correlated to the severity of coronary atherosclerosis and should therefore be the focused therapy target in patients with coronary artery disease.</p>

Fifteen Cases with Severe Combined Immunodeficiency Disease / 实用儿科临床杂志

2 g/L can′t excluded SCID.

Clinical features of patients with atypical coronary artery spasm / 中华心血管病杂志

<p><b>OBJECTIVE</b>This study is aimed to compare the clinical characteristics of patients with typical and atypical coronary artery spasm.</p><p><b>METHODS</b>Out of 64 patients with chest pain at rest and without significant coronary artery stenosis, coronary artery spasm was provoked by intracoronary injection of acetylcholine in 46 patients, including 12 with ST segment elevation (typical coronary artery spasm group) and 34 without ST segment elevation (atypical coronary artery spasm group). The demographic data, coronary angiographic findings, treadmill electrocardiogram, dipyridamole and rest thallium-201 myocardial perfusion computed tomography, and the follow-up clinical data of the two groups were compared.</p><p><b>RESULTS</b>The patients with typical coronary artery spasm were younger (47 +/- 6 vs. 58 +/- 12, P < 0.05) than patients with atypical coronary artery spasm group. Hyperlipidemia were more common in atypical coronary artery spasm group (74% vs. 33%, P < 0.05) and myocardial bridging was more common in patients with typical coronary artery spasm group (67% vs. 32%, P < 0.01). Focal coronary spasm during acetylcholine provocation was seen in 92% patients with typical coronary spasm and in 32% patients with a atypical coronary artery spasm (P < 0.01) while diffuse coronary spasm was seen in 8% patients with typical coronary spasm and in 68% patients with a atypical coronary artery spasm (P < 0.01). All patients with coronary artery spasm were treated with aspirin, calcium channel blockers, long-acting nitroglycerine, with or without lipid-lowering drugs, 2 patients with typical coronary spasm and 4 patients with atypical coronary spasm were rehospitalized due to chest pain and rest of the patients remained free of chest pain during the median follow-up period of 18 +/- 14 months.</p><p><b>CONCLUSION</b>Atypical coronary artery spasm is common in patients with rest angina and diffuse coronary microvascular spasm might be the cause of chest pain in these patients.</p>

Demonstration of a mechanism of anti-inflammatory effect of erythromycin on allergic airway inflammation in rat / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate whether erythromycin exerts anti-inflammatory effect on allergic airway inflammation and whether erythromycin modulate allergic airway inflammation by inhibiting nuclear factor kappa B (NF-kappa B) activation.</p><p><b>METHODS</b>Ovalbumin (OVA) together with aluminum hydroxide and Bordetella Pertussis was injected intraperitoneally to immunize SD rats and two weeks later 1% OVA was inhaled to challenge them for consecutive 7 days to mimic allergic airway inflammation. In treatment group, erythromycin was given orally (180 mg/kg.d) during the course of allergen exposure. WBC counts in bronchoalveolar lavage fluid (BALF) and lung specimen analysis were used to describe lung tissue inflammation. The expression of NF-kappa B subunit p65 in cell nucleus of lung tissue was measured by immunohistochemistry and NF-kappa B binding activation in lung tissue by electrophoresis mobility shift assay (EMSA).</p><p><b>RESULTS</b>Lung tissue specimen analysis indicated that the severity of allergic inflammation was reduced in treatment group. The number of total WBC in BALF (x 10(8)/L) (31 +/- 22) was lower than that in model group animals (66 +/- 28), P < 0.01. The number (x 10(3)/mm(2)) of cells with nuclear staining of NF-kappa B per square millimeter of submucosal region around large bronchus (1.4 +/- 0.4) was lower than that in model group animals (2.6 +/- 0.6), P < 0.01. NF-kappa B binding activity (32 +/- 14) was lower than that of model group (46 +/- 17), P < 0.05.</p><p><b>CONCLUSION</b>Erythromycin had an obvious protective role in allergic airway inflammation. Erythromycin inhibited NF-kappa B transcriptional activity to exert anti-inflammatory effect.</p>