Research progress on processing historical evolution, chemical constituents, and pharmacological action of Bombyx Batryticatus / 中国中药杂志

Bombyx Batryticatus is a precious traditional Chinese animal drug commonly used in clinical practice in China, which has the effects of extinguishing wind, stopping convulsions, dispelling wind, relieving pain, resolving phlegm, and dissipating mass. The processing of Bombyx Batryticatus has a long history. As early as in the Liu Song period of the Southern and Northern Dynasties, there was a record of the processing of Bombyx Batryticatus with rice swill. In addition to the processing with bran, honey bran, and ginger juice, which are still used today, there are also processing methods such as rendering, flour processing, wine processing, salt processing, oil processing, charcoal, and red dates processing in ancient times. After processing, the fishy smell of Bombyx Batryticatus can be removed, and avoid nausea and vomiting caused by the direct taking. Furthermore, processing can also facilitate the removal of surface hairs and toxicity reduction, making the medicinal material crispy and easy to crush. Previous studies have shown that the main chemical constituents of Bombyx Batryticatus include protein polypeptides, sterols, and flavonoids, with anticonvulsant, anticoagulation, antithrombotic, anti-cancer, hypnotic, hypoglycemic, and other pharmacological effects. This paper reviewed the processing historical evolution, chemical constituents, and pharmacological effects of Bombyx Batryticatus to lay a foundation for the research on the processing mechanism, quality control, and active core substances of Bombyx Batryticatus.

Effect of different electroacupuncture stimulation parameters on sudden hearing loss / 中国针灸

OBJECTIVE@#To compare the efficacy of different electroacupuncture (EA) frequencies and wave patterns combined with medication and medication alone for sudden hearing loss (SHL), and to explore better electroacupuncture stimulation parameters.@*METHODS@#All of 118 patients with SHL were randomly divided into an acupuncture and medication group 1 (group 1, 30 cases, 1 case dropped off), an acupuncture and medication group 2 (group 2, 30 cases), an acupuncture and medication group 3 (group 3, 31 cases) and a medication group (27 cases, 1 case dropped off ). The patients in the medication group were treated with conventional medication. On the base of the medication group, the patients in the group 1, 2, and 3 were treated with acupuncture at Ermen (TE 21), Tinggong (SI 19), Tinghui (GB 2), Fengchi (GB 20), etc. on the affected side, and EA at Ermen (TE 21)-Yifeng (TE 17), Tinghui (GB 2)-Yifeng (TE 17) alternately. The 3 groups were given continuous wave with frequency of 2 Hz, continuous wave with frequency of 50 Hz, and disperse-dense wave with frequency of 2 Hz/50 Hz respectively. The treatment was given once a day, 10 days were as one course, with 2 courses in total. Before and after treatment, the pure tone hearing threshold test was performed, and the curative effect of pure tone hearing threshold test and the curative effect of tinnitus, ear fullness and dizziness were compared in the 3 groups.@*RESULTS@#After treatment, the pure tone hearing threshold test values of each group were lower than those before treatment (@*CONCLUSION@#On the basis of conventional medication treatment, the addition of electroacupuncture can effectively improve the hearing and ear stuffiness symptoms of patients with SHL, and the disperse-dense wave with frequency of 2 Hz/50 Hz is more effective.

Effect of dhfr gene overexpression on ethanol-induced abnormal cardiovascular development in zebrafish embryos / 中国当代儿科杂志

OBJECTIVE@#To study the effect of dhfr gene overexpression on ethanol-induced abnormal cardiac and vascular development in zebrafish embryos and underlying mechanisms.@*METHODS@#dhfr mRNA was transcribed in vitro and microinjected into zebrafish fertilized eggs to induce the overexpression of dhfr gene, and the efficiency of overexpression was verified. Wild-type zebrafish were divided into a control group, an ethanol group, and an ethanol+dhfr overexpression group (microinjection of 6 nL dhfr mRNA). The embryonic development was observed for each group. The transgenic zebrafish Tg (cmlc2:mcherry) with heart-specific red fluorescence was used to observe atrial and ventricular development. Fluorescence microscopy was performed to observe the development of cardiac outflow tract and blood vessels. Heart rate and ventricular shortening fraction were used to assess cardiac function. Gene probes were constructed, and embryo in situ hybridization and real-time PCR were used to measure the expression of nkx2.5, tbx1, and flk-1 in the embryo.@*RESULTS@#Compared with the ethanol group, the ethanol+dhfr overexpression group had a significant reduction in the percentage of abnormal embryonic development and a significant increase in the percentage of embryonic survival (P<0.05), with significant improvements in the abnormalities of the atrium, ventricle, outflow tract, and blood vessels and cardiac function. Compared with the control group, the ethanol group had significant reductions in the expression of nkx2.5, tbx1, and flk-1 (P<0.05), and compared with the ethanol group, the ethanol+dhfr overexpression group had significant increases in the expression of nkx2.5, tbx1, and flk-1 (P<0.05), which were still lower than their expression in the control group.@*CONCLUSIONS@#The overexpression of the dhfr gene can partially improve the abnormal development of embryonic heart and blood vessels induced by ethanol, possibly by upregulating the decreased expression of nkx2.5, tbx1, and flk-1 caused by ethanol.

Effect of glucose metabolism on hypothalamus-pituitary-thyroid axis / 医学研究生学报

Glucose is the basic metabolic substrate and main source of energy for the body. A complex neuroendocrine system regulate blood glucose concertration to keep it at a constant level. Thyroid hormones participate in the regulation of glucose metabolism through the PI3K/ERK and other signaling pathways. At the same time, abnormal glucose metabolism can cause the abnormal regulation of hypothalamus pituitary thyroid axis. In this paper, we review the change and molecular mechanism of thyroid hormone, thyroid stimulating hormone and thyroid releasing hormone in Hypoglycemia and Hyperglycemia.

Effects of CXC-chemokine Receptor 7 Expression in Endothelial Progenitor Cells on Angiogenesis after Cerebral Ischemia-reperfusion Injury / 中国康复理论与实践

Objective:To explore the effect of upregulating CXC-chemokine receptor 7 (CXCR7) in endothelial progenitor cells (EPCs) on angiogenesis after cerebral ischemia-reperfusion injury. Methods:EPCs were isolated and cultured from human umbilical cord blood and identified. Then, the EPCs were transfected with CXCR7 overexpression lentiviral vector, and the expression of CXCR7 was identified with real-time PCR and Western blotting. The tube-like structure formation and apoptosis of EPCs under oxidized low density lipoprotein (ox-LDL) were detected with tube-like structure formation test and Annexin V/PI staining. Cerebral ischemia-reperfusion injury model in rats was established, and the qualified model rats were randomly divided into three groups after 24 hours reperfusion: PBS group (n = 12) was injected with phosphate buffers through tail vein, control group (n = 12) was injected the EPCs infected with control lentiviral vector, and CXCR7 group (n = 12) was injected with EPCs infected with CXCR7 overexpression lentiviral vector. Neurological function scores were determined seven and 14 days after transplantation. The cerebral infarct volume was measured, the number of GFP-positive cells in the ischemic site and the density of capillary were observed. Results:The expression of CXCR7 in EPCs increased after transfection (P < 0.01). Overexpression of CXCR7 improved tube formation and reduced apoptosis of EPCs under ox-LDL (P < 0.05). Compared with PBS and control groups , neurological function improved in CXCR7 group, with less infarct volume, more GFP-positive cells and density of capillary (P < 0.05). Conclusion:Up-regulating CXCR7 can improve the survival and angiogenesis of EPCs, and improve the repair of cerebral ischemia-reperfusion injury.

Cribriform adenocarcinoma of lung:a clinicopathological analysis of twenty cases / 临床与实验病理学杂志

Purpose To investigate the clinicopathological features,diagnosis and differential diagnosis,and genetic alteration of lung cribriform adenocarcinoma (LCA).Methods Clinicopathological features and immunophenotype were retrospectively evaluated in twenty LCA cases collected from Fujian Provincial Hospital,combined with genetic mutation analysis of EGFR.Results The tumors occurred in 5 women and 15 men aged from 39 to 74 years (median =59.25).Diameter of masses ranged from 1.5-5.5 cm (median =3.265).Histologically,the tumors were unencapsulated and were divided by fibrous septa into lobules.Major parts of the lesions were composed of areas with solid and microcystic growth patterns.The most striking cytological feature was that the tumor cell nuclei were pale staining with a ground glass feature,and they often appeared to overlap.Immunephenotypically,the tumor cells were positive for TTF-1(20/20),Napsin A (18/20),SPB(19/20),CK7(20/20),CD56 (2/20),EGFR (7/20),β-catenin (4/20),ALK (2/20).Tg,CDX-2,Syn,CgA,S-100 and SMA were negative.Gene mutation detection confirmed that EGFR genes were mutating in nine cases,and eleven cases were wild type.The twenty cases were all performed for the completely surgical resection.20 cases accepted chemotherapy,and 4 cases unaccepted chemotherapy.Conclusion LCA is a rare tumor with distinct morphologic feature.Clinically and pathologically,it needs to differentiate from alveolus adenocarcinoma,adenoid cystic carcinoma and papillary carcinoma of the thyroid,and so on.The primary treatment for LCA is completely surgical excision and chemotherapy.Otherwise its prognosis is poor.

Betanin down-regulates the protein coding transcripts of the pyruvate by-pathway in mitochondia of HeLa cells / 中国药学杂志

OBJECTIVE: To study the effects of batanin on proliferation of HeLa cells and the relationships with pyruvate metabolism and glycolysis. METHODS: The HeLa cells were cultured in vitro and betanin at 0, 62.5 and 500 mg·L-1 was added after the attachment of the cells. The cells were harvested after 48 h of culture for the cell counting and storaged at low temperature. Referring to human genome, the frozen cells were carried out for the analysis and calculation of high-throughout transcrip-tome. Utilizing internet data base, the parts related with glycolysis, pyruvate metabolism and citric cycle were selected and analyzed to study the effect of betanin on the protein coding transcripts of the glycolysis and citric cycle in HeLa cells. RESULTS: Betanin with both of the 2 concentrations inhibited the proliferation of the cultured HeLa cells, and affected the transcript reads of the cultured cells. There were 74 of protein coding transcripts expressed in HeLa cells, but only 40 of them had the reads >1 or <1 but with significant difference between treatments. Betanin significantly up-regulated the protein coding transcripts of the phosphofructokinase (platelet) (ENST00000381075) and fructose-1,6-bisphosphatase 1 (ENST00000375326) which were rate-limited enzymes in the glycolysis and gluconeogenesis respectively, it was benefit for cell metabolism, possibly being the stress response of the cell to betanin. However, betanin significantly down-regulated the protein coding transcript of the mitochondial lactate dehydrigenase D (LDHD) (ENST00000450168) of HeLa cell, and the mitochondial phosphoenolpyruvate carboxykinase 2 (PEPCK) also had a tendency to being down-regulated, which all were related with pyruvate metabolism. CONCLUSION: The inhibition of betanin on proliferation of HeLa cell is related with the down-regulation of the protein coding transcripts of the LDHD and PEPCK of the mitochondrial pyruvate metabolism of the cell.

Two-dimensional electrophoresis analysis for protein profile change in zebrafish alcohol syndrome model / 中华围产医学杂志

Objective To study the putative mechanisms underlying fetal alcohol syndrome by comparative protein-profile analysis between normal and ethanol-treated zebrafish embryo with twodimensional electrophoresis (2-DE).Methods Zebrafish embryos were exposed in 400 mmol/L ethanol at dome stage for 3 hours,and then ethanol-induced abnormalities were observed.Proteomes of zebrafish embryos at early stages including zygote stage,dome stage,shield stage and 5-somite stage,were separated by 2-DE.The subtraction analysis method was applied to eliminate the interference from maternal derived proteins.The ethanol-treated embryos at 5-somite stage was analyzed by 2-DE,and the protein profile was compared with that generated from control embryos at the same stage.The data obtained from 2-DE analysis were verified by in-situ hybridization.Results 400 mmol/L ethanol treatment caused axial malformation (62%) and cyclopia (60%) in zebrafish embryos.The 2-DE analysis showed that the expression of Collagen2al (Col2a1) and TAR DNA binding protein (TDP) was decreased in 12 hours post fertilization (12 hpf) ethanol-treated embryos by 81% and 73%,respectively.The in-situ hybridization also demonstrated that the expression of Col2al in axial mesoderm was reduced by ethanol treatment at the same stage.But for 24 hpf ethanoltreated embryos,the expression of Col2al in axis recovered to a comparable level to that in control embryos,while the structure of neural tube was disrupted severely by ethanol exposure.Conclusions It is suggested that the expressions of Col2al and TDP were disrupted by ethanol during early stage,which might induce the zebrafish developmental abnormalities.The ethanol interference on early expression of Col2al is supposed to be one of the major reasons leading to later abnormalities of axis and neutral tube.

Effects of folic acid on the development of heart of zebrafish / 中华儿科杂志

<p><b>OBJECTIVE</b>To construct the folic acid deficient model in zebrafish and observe the abnormal cardiac phenotypes, to find the optimal period for supplementing folic acid that can most effectively prevent the heart malformation induced by folic acid deficiency, and to investigate the possible mechanisms by which folic acid deficiency induces malformations of heart.</p><p><b>METHOD</b>The folic acid deficient zebrafish model was constructed by using both the folic acid antagonist methotrexate (MTX) and knocking-down dhfr (dihydrofolate reductase gene). Exogenous tetrahydrofolic acid rescue experiment was performed. Folic acid was given to folic acid deficient groups in different periods. The percent of cardiac malformation, the cardiac phenotypes, the heart rate and the ventricular shortening fraction (VSF) were recorded. The out flow tract (OFT) was observed by using fluorescein micro-angiography. Whole-mount in situ hybridization and real-time PCR were performed to detect vmhc, amhc, tbx5 and nppa expressions.</p><p><b>RESULT</b>About (78.00 ± 3.74)% embryos in MTX treated group and (68.00 ± 6.32)% embryos in dhfr knocking-down group had heart malformations, including the abnormal cardiac shapes, the hypogenesis of OFT and the reduced heart rate and VSF. Giving exogenous tetrahydrofolic acid rescued the above abnormalities. Given the folic acid on 8 - 12 hours post-fertilization (hpf), both the MTX treated group (20.20% ± 3.77%) and dhfr knocking-down group (43.40% ± 4.51%) showed the most significantly reduced percent of cardiac malformation and the most obviously improved cardiac development. In folic acid deficient group, the expressions of tbx5 and nppa were reduced while the expressions of vmhc and amhc appeared normal. After being given folic acid to MTX treated group and dhfr knocking-down group, the expressions of tbx5 and nppa were increased.</p><p><b>CONCLUSIONS</b>The synthesis of tetrahydrofolic acid was decreased in our folic acid deficient model. Giving folic acid in the middle period, which is the early developmental stage, can best prevent the abnormal developments of hearts induced by folic acid deficiency. Folic acid deficiency did not disrupt the differentiations of myosins in ventricle and atrium. The cardiac malformations caused by folic acid deficiency were related with the reduced expressions of tbx5 and nppa.</p>

Effects on function rehabilitation of vocal cord after vocal cord polyps surgery treated with acupuncture at Sheng's four points of throat / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects on function rehabilitation of vocal cord after vocal cord polyps surgery treated with acupuncture at Sheng's Four Points of Throat.</p><p><b>METHODS</b>Sixty cases were randomly divided into a combined therapy group of Four Points of Throat and medication (group 1, 30 cases) and a medication group (group 2, 30 cases). In group 1 Four Points of Throat were punctured and routine medication was applied; in group 2, only routine medication was applied. The scores of symptom-sign and status of voice were observed and analyzed before and after treatment in two groups.</p><p><b>RESULTS</b>At 4th day after the surgery, the improvement of the symptom-sign scores in group 1 was more significant than that in group 2 (P < 0.05); and the voice analysis status in group 1 was superior to that in group 2 (all P < 0.05). The total effective rate was 83.3% (25/30) in group 1, which was superior to that of 60.0% (18/30) in group 2 (P < 0.05).</p><p><b>CONCLUSION</b>The effect on function rehabilitation of vocal cord after vocal cord polyps surgery treated with the combined therapy group of Four Points of Throat and routine medication is favorable, superior to that with routine medication therapy.</p>

Facial nerve function index evaluation on deviation of the mouth in intractable facial palsy treated with sticking needle and traction method on three points of the mouth / 中国针灸

<p><b>OBJECTIVE</b>To probe into the effective acupuncture technique for deviation of the mouth in intractable facial palsy.</p><p><b>METHODS</b>One hundred and one cases of intractable facial palsy were randomly divided into an observation group (48 cases) and a control group (53 cases). Cuanzhu (BL 2), Sibai (ST 2), Jiache (ST 6) and Qianzheng (Extra) on the affected side were punctured in two groups. Additionally, three acupoints of the mouth were supplemented, named Dicang (ST 4), Kouheliao (LI 19) and Jiachengjiang (Extra) were added, and the sticking needle and traction method was adopted on them in observation group. the routine needling technique was applied in control group. The treatment was given once a day and 10-day treatment made one session. The changes in facial nerve function index (FNFI) were observed in 2 sessions of treatment.</p><p><b>RESULTS</b>After treatment, FNFI in two groups increased significantly (both P < 0.01), but the improvement in observation group was better than that in control group (P < 0.01). In observation group, the basic recovery rate of FNFI was 87.5% (42/48), which was higher than that (67.9%, 36/53) in control group (P < 0.05).</p><p><b>CONCLUSION</b>The sticking needle and traction method o three points is the quite effective approach in the treatment of deviation of the mouth in intractable facial palsy.</p>

The experimental research for contribution of insulin-like growth factor binding protein-2 during zebrafish embryonic cardiovascular development / 复旦学报（医学版）

Objective To establish a zebrafish IGFBP-2 gene knock-down model by morphilino modified antisense oligonucleotide injection, so as to investigate the abnormal phenotypes of heart and vessels in early stage of zebrafish development and the expression of zebrafish cardiogenesis related genes. Methods The spatiotemporal expression of IGFBP-2 gene in early stage of zebrafish development was testified by whole mount in situ hybridization with antisense RNA IGFBP-2 probe. The IGFBP-2 morpholino (IGFBP-2 MO) that especially inhibited the gene promoter and standard control morpholino (Con-MO) were designed and synthesized by Gene-tools Corporation. Four different concentration gradients (0.05, 0.10, 0.25 and 1.0 mmol/L) were set as IGFBP-MO injection groups with 0.25 mmol/L Con-MO injection group and wild type group as controls. Contribution to the incidence of heart abnormal phenotypes and mortality rate induced by 4 different IGFBP-2 concentrations injection group was recorded and compared with 2 control groups. Heart abnormal phenotypes at different developmental stages in 0.25 mmol/L IGFBP-2 injection group were observed in detail. To validate the effectiveness of IGFBP-2 MO, the expression of enhanced green fluorescence presented by wild type zebrafish embryos at 12hpf which received single injection of IGFBP-2 EGFP recombinant plasmid and those co-injected with Con-MO or IGFBP-2 MO were detected. To investigate the regulation relationship between IGFBP-2 gene and other cardiogenesis related genes, expression of atrium specific marker gene Amhc was detected in IGFBP-2 MO and wild type group by in situ hybridization. Ventricle specific green fluorescence of Vmhc-EGFP transgenic zebrafish embryos whose IGFBP-2 gene was knocked-down were compared with those untreated. Zebrafish peripheral vascular development in the IGFBP-2 MO group was also checked out by micro-angiography. Results Whole mount in situ hybridization revealed that IGFBP-2 gene expressed in turn at eyes, midbrain and then focused on liver in early stage of zebrafish development. The micro-injection of 0.25 mmol/L IGFBP-2 MO resulted in heart malformation in nearly 60% of all injected zebrafish embryos. Heart malformation phenotypes included slow heart beat, pericardial edema, weak ventricle systole contraction and heart tube looping disorder. Some of them represented atria dilation, blood regurgitation and ciculation obstruction. Wild type zebrafish embryos that received single injection of IGFBP-2 EGFP plasmid DNA or co-injected with Con-MO presented strong enhanced green fluorescence at 12hpf, meanwhile, the fluorescence was barely seen in the embryos co-injected with IGFBP-2 MO. This strongly validated the gene specific knock-down effect of IGFBP-2 MO. Amhc was down-regulated at 48hpf in IGFBP-2 MO group. Vmhc-EGFP transgenic zebrafish down-regulated by IGFBP-2 gene also resulted in attenuated expression of ventriclar-specific green fluorescence protein at 48hpf. Intersegmental blood vessels of IGFBP-2 MO group by micro-angiography at 60hpf demonstrated an sparsate and chaos image, which suggested that IGFBP-2 gene expression was involved in the regulation of normal vascular development. Conclusions Micro-injection of IGFBP-2 MO is an efficient way to knock-down IGFBP-2 gene in zebrafish embryos. IGFBP-2 gene expression down-regulation leads to heart and vessels maldevelopment and have an impact on the expression of cardiogenesis related genes of zebrafish embryos as well. In short, IGFBP-2 plays a critical role in the normal cardiovascular development of zebrafish embryos.

Effect of Tbx1 knock-down on cardiac performance in zebrafish / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Tbx1 is the major candidate gene for DiGeorge syndrome (DGS). Similar to defects observed in DGS patients, the structures disrupted in Tbx1(-/-) animal models are derived from the neural crest cells during development. Although the morphological phenotypes of some Tbx1 knock-down animal models have been well described, analysis of the cardiac performance is limited. Therefore, myocardial performance was explored in Tbx1 morpholino injected zebrafish embryos.</p><p><b>METHODS</b>To elucidate these issues, Tbx1 specific morpholino was used to reduce the function of Tbx1 in zebrafish. The differentiation of the myocardial cells was observed using whole mount in situ hybridization. Heart rates were observed and recorded under the microscope from 24 to 72 hours post fertilization (hpf). The cardiac performance was analyzed by measuring ventricular shortening fraction and atrial shortening fraction.</p><p><b>RESULTS</b>Tbx1 morpholino injected embryos were characterized by defects in the pharyngeal arches, otic vesicle, aortic arches and thymus. In addition, Tbx1 knock down reduced the amount of pharyngeal neural crest cells in zebrafish. Abnormal cardiac morphology was visible in nearly 20% of the Tbx1 morpholino injected embryos. The hearts in these embryos did not loop or loop incompletely. Importantly, cardiac performance and heart rate were reduced in Tbx1 morpholino injected embryos.</p><p><b>CONCLUSIONS</b>Tbx1 might play an essential role in the development of pharyngeal neural crest cells in zebrafish. Cardiac performance is impaired by Tbx1 knock down in zebrafish.</p>

Arthroscopic release to treat subtalar stiffness after calcaneal fracture / 中华外科杂志

<p><b>OBJECTIVE</b>To report the results of arthroscopic release to treat subtalar stiffness after calcaneal fractures.</p><p><b>METHODS</b>From September 2004 to December 2006, 10 cases of subtalar stiffness were treated. There were 8 male and 2 female cases, with an average age of 36 years old (ranging from 18 to 48). All, but 2 cases, had single subtalar involvement. The routine triple portals (lateral, anterolateral, posterolateral portals) were applied with the patient placed in the lateral decubitus position. The anterior capsule, lateral gap, calcaneofibular ligament, posterior capsule together with the posteromedial corner of the subtalar joint were released step by step under arthroscopic control. Finally, manual release was performed.</p><p><b>RESULTS</b>All cases were followed-up for 12 to 36 months (mean, 24.5 months). According to the AOFAS hindfoot activity rating scale, 10 cases were rated as Grade III, 2 as Grade II before the surgery. Nine cases were improved to Grade I, 3 to Grade II at the last follow-up after the surgery. AOFAS hindfoot scores were significantly improved from 71.4 before the surgery to 90.6 at the last follow-up (P < 0.01). All cases returned to the previous work at an average of 1.8 months (range, 1 to 3 months) after the surgery.</p><p><b>CONCLUSION</b>Arthroscopic release to treat subtalar stiffness after calcaneal fracture has such advantages as minimally-invasiveness, simplicity and effectiveness.</p>

Comparison of indirect immunofluorescence assay and ELISA for detecting antinuclear antibodies and anti-double-stranded DNA antibodies / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare indirect immunofluorescence assay (IIFA) and enzyme-linked immunosorbent assay (ELISA) for detecting antinuclear antibodies (ANA) and anti-double-stranded DNA antibodies (anti-dsDNA).</p><p><b>METHODS</b>A total of 125 serum samples were obtained from patients with established or suspected autoimmune disease, and 82 samples were used for ANA detection and 57 for anti-dsDNA detection using both IIFA and ELISA. Fourteen samples were examined for both ANA and anti-dsDNA. In cases where discrepancy occurred in the results by the two methods, extractable nuclear antigens were detected using immunoblotting.</p><p><b>RESULTS</b>The positivity rate of ANA detected by IIFA and ELISA was significantly different (87.8% and 73.17%, respectively, P<0.01), but the positivity rate of anti-dsDNA was similar between IIFA and ELISA (77.19% and 71.93%, respectively, P>0.05). The percent agreement between the two testing methods with different cutoff values of ANA and anti-dsDNA showed significant differences (P<0.01), and for some uncommon patterns, the percent agreement of the two methods was lowered in ANA detection but remained unchanged for anti-dsDNA with different ANA patterns. High percent agreements of the two methods were obtained with the cutoff ANA titer of 1:100 and the cutoff anti-dsDNA value of weak positivity, but they demonstrated a significant difference in testing low-titer ANA and anti-dsDNA.</p><p><b>CONCLUSION</b>IIFA is more sensitive than ELISA in detecting the total ANA and anti-dsDNA. ELISA prescreening combined with IIFA can obtain the information of the nuclear pattern and allow the observation of the titer alterations. The combination of two or more testing methods can greatly enhance the accuracy of the results.</p>

The Effects of BHC80 Down Regulation on Embryonic Erythropoiesis in Zebrafish / 生物化学与生物物理进展

Whole mount in situ hybridization with BHC80 RNA probe showed that BHC80 was expressed in zebrafish central nervous system. Morpholino-modified antisense oligonucleotide was injected into zebrafish zygotes to knock down BHC80 expression. BHC80 knockdown resulted in striking decrease of erythrocytes and accumulation of erythrocytes at PBI. Further investigation of embryonic erythrocytes marker βe3 globin and hematopoiesis transcription factors gata1, c-myb and lmo2 by in situ hybridization showed that the erythroid progenitors marked with gata1 in BHC80 knockdown embryos were high proliferation and their differentiation were delayed, which led to decrease of erythrocytes and accumulation of erythrocytes at PBI. Both in situ hybridization and microangiography indicated that vasculature pattern of BHC80 knockdown embryos were almost normal.

The effects of folic acid deficiency on the axial development of zebrafish / 复旦学报（医学版）

Objective To construct a folic acid deficient model in zebrafish and to observe the axial development in folic acid deficient embryos, so as to probe the mechanism by which folic acid deficiency induces abnormal development of axis. Methods We constructed the folic acid deficient zebrafish model by both using the antagonism of dihydrofolate reductase (MTX) and knocking-down dihydrofolate reductase gene. Then we observed the axial excursion of folic acid deficient embryos at 17 hpf under microscope. We labeled and observed the positions of liver, spleen and heart by using whole-mount in situ hybridization with specific antisense RNA probes. The expressions of some genes, which are down stream factors of Nodal signal pathway and important for axial development, were detected by whole-mount in situ hybridization and Real-time PCR. Results Parts of folic acid deficient embryos had axial excursion and abnormal positions of liver, spleen and heart. The expressing intensities of ntl and gsc appeared normal in folic acid deficient embryos, but the expressing spatial patterns were abnormal, which revealed the malformation of axial mesoderm. Conclusions Folic acid deficiency induced the abnormal development of axis and the malformation of axial mesoderm. Folic acid deficiency had no obviouse effect on Nodal pathway.

Epidemiological study of the effects of smoking cigarette on thyroid gland / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the effect of cigarette smoking on thyroid gland volume, thyroid function and thyroid autoantibodies in the areas with different iodine intakes.</p><p><b>METHODS</b>A cross-sectional epidemiological study in Panshan (mild iodine-deficient area), Zhangwu (more than adequate iodine intake area) and Huanghua (iodine-excessive area) was conducted in 3761 subjects in 1999.80.2 % of them were followed up in 2004. Questionnaires, thyroid function, thyroid autoantibodies, urinary iodine concentration,and thyroid B ultrasound were performed.</p><p><b>RESULTS</b>The prevalence of goiter was higher in smokers than in non-smokers (15.1% vs. 11.5%, P< 0.05). The average thyroid volume was higher in smokers with phenomenon more obvious in Panshan and Huanghua areas. Data from logistic analysis showed that smoking cigarette was an independent risk factor of goiter. There was no difference in serum TSH and Tg level between smokers and non-smokers. The positive rate of TPOAb (>100 IU/ml) was higher in smokers than in non-smokers(10.8% vs. 9.0 % , P <0.05) and was especially obvious in Huanghua area. Smoking was a independent risk factor of increasing positive rate of TPOAb. During the prospective observation,it was found that the incidence of positive TPOAb(>,100 IU/ml) was 7.4% in the subjects that were from non-smokers turning to smokers and 2.9% in those whose smoking behavior did not change. Logistic analysis indicated that the shifting from non-smoking to smoking was independent risk factor for the increase on high incidence of positive TPOAb.</p><p><b>CONCLUSION</b>Smoking cigarette was a independent risk factor of goiter. Smoking was also a risk factor of increasing TPOAb positive rate. Shifting from not smoking to smoking was an independent risk factor of increasing high incidence of positive TPOAb.</p>

Effect of dihydrofolate reductase gene knock-down on the expression of heart and neural crest derivatives expressed transcript 2 in zebrafish cardiac development / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Folic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of folic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase (DHFR) gene knock-down zebrafish embryos, investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 (HAND2) and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations.</p><p><b>METHODS</b>Morpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression.</p><p><b>RESULTS</b>DHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos (P < 0.05). Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos.</p><p><b>CONCLUSIONS</b>DHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation.</p>

Folic acid antagonist methotrexate causes the development malformation of heart and down-regulates the BMP2b and HAS2 expressions in zebrafish / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of methotrexate (MTX), a folic acid antagonist which can lead to folic acid deficient, on the cardiac development and on the expressions of BMP2b and HAS2 in zebrafish.</p><p><b>METHODS</b>The zebrafish embryos at 6-48 hrs post fertilization (hpf) were treated with various concentrations of MTX (0.5 x 10(-3), 1.0 x 10(-3) and 2.0 x 10(-3) M). At 48 hpf, the percentage of cardiac malformation and heart rate were recorded. The zebrafish embryos at 6-10 hpf treated with 1.5 x 10(-3) M MTX were used as the MTX treatment group. At 24 and 48 hpf the cardiac morphology was observed under a microscope. The expressions of BMP2b and HAS2 in zebrafish were detected by in situ antisense RNA hybridization and real-time PCR.</p><p><b>RESULTS</b>6-12 hpf, the early embryonic developmental stage, was a sensitive period that MTX affected cardiac formation of zebrafish. The retardant cardiac development and the evidently abnormal cardiac morphology was found in the MTX treatment group. The results of in situ antisense RNA hybridization showed that the expressions of BMP2b and HAS2 in the zebrafish heart were reduced in the MTX treatment group at 36 and 48 hpf. The real-time PCR results demonstrated that the BMP2b expression decreased at 12, 24, 36 and 48 hpf, and that the HAS2 expression decreased at 24, 36 and 48 hpf in the treatment group compared with the control group without MTX treatment.</p><p><b>CONCLUSIONS</b>The inhibition of folic acid function may affect cardiac development of early embryos, resulting in a retardant development and a morphological abnormality of the heart in zebrafish, possibly by down-regulating the expressions of BMP2b and HAS2.</p>