Eating behaviors and growth status in children with attention deficit and hyperactivity disorder / 中国学校卫生

Objective@#To discuss the correlation between growth status and eating behaviors in children with attention deficit and hyperactivity disorder (ADHD), providing reference data for management and dietary behavior guidance among ADHD children.@*Methods@#A total of 703 children aged 4-13 years old were collected from the ADHD patients from Children s Health Department of Children s Hospital of Nanjing Medical University from June to September, 2019. The demographic characteristics and information regarding children’s eating behaviors were collected by self-designed questionnaire and Chinese version of the parent-completed Children’s Eating Behavior Questionnaire(CEBQ). The correlation physical growth with dietary behaviors among the ADHD children were analyzed.@*Results@#Food avoidant behaviors, including satiety responsiveness, slowness in eating and emotional undereating in ADHD children with thinness scored significantly higher than that of children with short stature, overweight and obesity(F=17.57, 29.32, 4.07, P<0.01), while food approach behaviors, including food responsiveness, enjoyment of food, desire to drink and emotional overeating scored higher in obese children, compared to other three groups(F=24.54, 47.44, 2.96,5.85, P<0.05). Multiple linear regression analysis showed that, after adjusting for the confounders, satiety responsiveness, slowness in eating were still negatively associated with BMI-Z score of the ADHD children(B=-0.05, -0.07, P<0.01). Food responsiveness, enjoyment of food and emotional overeating had a positive association with the BMI-Z score(B=0.04, 0.09, 0.05, P<0.05).@*Conclusion@#Emotional eating and high food responsiveness in ADHD children are associated with the overweight and obesity, while long eating time and high satiety responsiveness is associated with underweight among ADHD children. For clinical doctors and parents, problematic eating behaviors among ADHD children should be concerned regarding its negative effects on growth and development, besides core symptoms of ADHD.

Analysis of a pedigree affected with congenital dysfibrinogenemia due to heterozygous Gln195Arg mutation of fibrinogen γ chain gene / 中华检验医学杂志

Objective:To analyze the phenotype and genotype of a Chinese pedigree with congenital dysfibrinogenemia and investigate the molecular mechanism of the disease.Methods:Pedigree analysis. Peripheral blood samples were collected from 7 members of the pedigree and routine coagulation tests were conducted. The activity of fibrinogen was measured using Clauss method, and fibrinogen antigen was measured by immunoturbidimetry. All the exons and exon-intron boundaries of FGA, FGB and FGG genes were amplified using PCR, which was followed by direct sequencing. Electrophoretic and immunological analysis of fibrinogen, fibrinogen clottability measurement, fibrin polymerization measurement and scanning electron microscopy were used to investigate the pathogenesis of this disease. Results:The proband showed normal activated partial thromboplastin time (APTT) , prolonged prothrombin time(PT), thrombin time (TT),and reptilase time (RT).The antigen level of fibrinogen in the proband (1.6 g/L) decreased slightly, while the activity level of fibrinogen (0.7 g/L) decreased significantly. His father and grandmother showed normal APTT and PT, prolonged TT and RT. The antigen levels of fibrinogen in both of them were normal (2.0 g/L and 2.2 g/L, respectively), while the activity levels of fibrinogen were low (1.0 g/L and 1.1 g/L, respectively). The results of other members from the pedigree were all within the normal range. Genetic analysis revealed a heterozygous A>G mutation at nucleotide 4774 in exon 6 of FGG gene in the proband, which was predicated to be a novel Gln195Arg mutation. The mutation was also found in his father and grandmother.Western blot results showed that no abnormal bands of plasma fibrinogen were found in the proband, his father and grandmother. The fibrinogen clottability in the proband was 49.3%, while that in the heathy control was 98.9%. Both thrombin-induced fibrin polymerization and reptilase-induced fibrin polymerization were significantly impaired in the proband, compared to that in the heathy control. Scanning electron microscopy revealed that compared with the heathy control, the average fiber diameters of the fibrin clot in the proband increased significantly ( P<0.001), while the density of fibers decreased and the arrangement of fibers was sparse. Conclusions:The heterozygous Arg19Gly mutation, which probably damages functions of fibrinogen, should be responsible for the congenital dysfibrinogenemia in this pedigree. This mutation has not been reported.

Identification and molecular pathogenesis study of a case of inherited dysfibrinogenemia / 临床检验杂志

Objective@#To analyze the phenotype and genotype of a Chinese pedigree with inherited dysfibrinogenaemia and investigate the molecular mechanism of the disease. @*Methods@#Venous blood samples were collected from all family members, and routine coagulation tests were conducted. Functional fibrinogen in venous blood samples was measured by Clauss method, and the antigen level of fibrinogen in plasma was measured by immunoturbidimetry assay. All the exons and exon-intron boundaries of the three fibrinogen genes were analyzed by direct sequencing. Fibrinogen electrophoresis, fibrinogen clottability measurement, fibrin polymerisation measurement and electron microscopy scanning were also used to investigate the molecular characteristics and pathogenesis. @*Results@#The proband had normal activated partial thromboplastin time, prothrombin time and plasma fibrinogen antigen, but prolonged thrombin time, prolonged reptilase time and reduced fibrinogen activity level, which were also found in his father. The sequencing results of the proband revealed heterozygous A1211G in the exon 2 of FGA gene originating from his father, which caused Arg19Gly missense mutation. The western-blot results showed that no abnormal bands of plasma fibrinogen were found in the proband and his father. Both thrombin-induced fibrin polymerisation and reptilase induced fibrin polymerisation were significantly impaired compared to normal control. Fibrinogen clottability measurement showed that only about 20.8% molecules of plasma fibrinogen in the proband were involved in the clot formation. Scanning electron microscopy revealed that the proband′s average fibre diameters were found to be significantly thicker than that of the control(P＜0.001), and the density was smaller than that of normal control. @*Conclusion@#The Arg19Gly mutation should be responsible for the proband′s dysfibrinogenaemia and the relevant clinical symptoms.

Exploration and practice of"5+3"integrated training model / 中华医学教育探索杂志

"5+3" integrated training program is a new training model to cultivate doctors under the background of preclinical teaching and clinical training integration. Based on practice in Shantou University Medical College, the program design current progress and solution to the problem were elaborated. It adheres to principles of overall optimization and orienting to clinic, and to ensure the smooth implementation of the training model. Which will provide important reference for the reform of long-term medical training program in higher medical colleges and universities.

Glutamine supplementation provides neuroprotection by inhibiting microglia-mediated neuroinflammatory response after traumatic brain injury in rats / 中华临床营养杂志

Objective To investigate the effects and mechanisms of Glutamine (Gln) supplementation on neurobehavioral outcome,neuronal apoptosis,microglia polarization,and neuroinflammatory response after traumatic brain injury (TBI) in rats.Methods TBI animal models were established using Feeney's method.Sixty Wistar rats were randomly divided into three groups:control group (Con),traumatic brain injury group (TBI),and glutamine supplementation group (TBI+Gln).We measured rat behavioral outcomes by modified neurologic severity score (mNSS) tests at day 1,3,7 and 14 after TBI.Apoptotic neurons were determined by Nissl staining.The microglia polarization relatived protein (Iba-1,CD16,CD86) expressions in TBI cerebral cortices were determined by immunohistochemistry staining and western blotting,respectively.While,the serum levels of tumor necrosis factor-α (TNF-α),interleukin (IL)-1 and interferon (IFN)-γ were tested by enzyme linked immunosorbent Assay (ELISA).Results Compared with the Con group,the levels of neurobehavioral outcome,neurons apoptosis,microglia polarization and neuroinflammatory factors were significantly increased in the other two groups (P=0.00).Compared with the TBl group,glutamin supplementation improvedneurobehavioral outcome [7 d:(10.74±0.25) points vs.(8.94±0.24) points,P=0.01;14 d:(8.77± 0.16) points vs.(7.43±0.13) points,P=0.03].Meanwhile,glutamin supplementation suppressed the apoptotic rates of neurons [3d:(80.18±8.38)％ vs.(65.47±7.02)％,P=0.01;7 d:(58.90±6.12)％ vs.(42.73±4.88)％,P=0.01;14d:(39.56±2.95)％vs.(31.12±3.16)％,P=0.01],inhibited protein expressions of Iba-1 and CD16,and increased the protein expression of CD86,which promoted the phenotypic shift of microglia from M1 to M2 phenotype,inhibited microglial activation,and thus reduced TBI-induced neuroinflammatory factors [TNF-α:(125.42 ± 12.81) pg/ml vs.(74.36 ± 9.25) pg/ml,P =0.01;IL-1:(69.04±8.48) pg/ml vs.(34.73±3.92) pg/ml,P=0.01;TNF-α:(89.75±9.40) pg/ml vs.(45.62±6.64) pg/ml,P=0.02].Conclusion Glutamine supplementation can markedly reduce neuron apoptosis and improve neurological outcomes after TBI,possibly mediated by promoting the phenotypic shift of microglia from M1 to M2 phenotype and thus reducing TBI-induced neuroinflammatory factors.

Therapeutic Efficacy of Buzhong Yiqi Decoction Containing Different Doses of Radix Astragali for Stage Ⅲ Internal Hemorrhoids Patients with Spleen Deficiency and Sinking of Qi and Its Effect of on Fibulin-3 Expression in Hemorrhoid Tissues / 广州中医药大学学报

Objective To compare the effect of Buzhong Yiqi Decoction (BYD) containing different doses of Radix Astragali on fibulin-3 expression in the hemorrhoid tissues of stage Ⅲ internal hemorrhoids patients with spleen deficiency and sinking of qi, and to evaluate its therapeutic efficacy and possible mechanism. Methods Fifty-five qualified patients were randomly divided into control group(N = 15), Chinese medicine group 1(N =20), and Chinese medicine group 2(N=20). All of the 3 groups were treated with operation, and additionally, Chinese medicine group 1 was given BYD containing Radix Astragali 20 g, and Chinese medicine group 2 was given BYD containing Radix Astragali 50 g orally after operation. The scores of anal pendant expansion and anal prolapse were evaluated, and the expression level of fibulin-3 in the hemorrhoid tissues was detected by Western blot method. Results After treatment, the symptoms of anal pendant expansion and anal prolapse were improved in the 2 Chinese medicine groups (P 0.05). Conclusion BYD with large dose of Radix Astragali exerts stronger therapeutic efficacy for the treatment of prolapsed hemorrhoids than BYD with small dose of Radix Astragali, and its therapeutic mechanism has no obvious relation with promoting the increase of fibulin-3 expression.

Association of oxygen uptake efficiency in exercise test with exercise capacity in patients of obstructive sleep apnea hypopnea syndrome / 中华全科医师杂志

Objective To explore the characteristics of oxygen uptake efficiency (OUES) in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) and analyze the relationship between OUE and severity of disease.Methods Pulmonary function test,polysomnogram and cardiopulmonary exercise testing were performed in 35 patients with OSAHS and 25 age-matched healthy volunteers.Their successive breathing respiratory exchange parameters were collected and analyzed.And t and x2 tests were used for 2 sample comparison.Correlation analysis was performed by Pearson correlation test.Results Significant differences in peak VO2 and peak VO2 ％ pred existed between OSAHS and normal control groups [(18±4) vs.(28 ±6) L/min,P＜0.01;(68±14) vs.(84±16) ％,P＜0.01].Compared with normal control group [(2.3 ±0.5) L · min-1 · lg-1 ; (36 ±4) ml/L; (36 ±4) ml/L],OUES,OUEP and OUE@AT of OSAHS group [(1.8 ± 0.4) L · min-1 · lg-1 ; (31 ± 5) ml/L; (30 ± 5) ml/L] were significantly lower (t =3.78-4.49,all P ＜0.01).And OUES,OUEP and OUE@AT in OSAHS patients were correlated (r =0.53-0.67,all P ＜0.01) positively with exercise tolerance (peak VO2％ pred) while negatively with apnea hypopnea index (AHI) (r=-0.67--0.54,all P ＜0.01).Conclusion The oxygen uptake efficiency of patients with OSAHS is significantly reduced compared to that of normal subjects.And it is correlated negatively with severity of disease.

Study of molecular mechanism and antigen expression of CisAB01 blood group / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the molecular mechanism of CisAB01 subtype in the ABO blood group system, and to investigate the expression of A and B antigens in red blood cells (RBCs).</p><p><b>METHODS</b>For 5 unrelated individuals with the CisAB phenotype, the molecular basis for the blood type was studied with serological assay, DNA sequencing and haplotype analysis. Bioinformatics analysis was carried out to investigate the changes in structure and function of relevant enzymes. Expression of A and B antigens in RBCs of CisAB01 was detected by flow cytometry.</p><p><b>RESULTS</b>All of the 5 samples were found to have a CisAB01 subtype. The underlying mutations, 467C>T and 803G>C in exon 7, have resulted in replacement of amino acid P156L and G268A. The mean fluorescence intensity (MFI) of A antigen in CisAB01 cases was 135, while the control group was 172. The B antigens in CisAB01 cases (MFI=38) showed significant decrease in MFI compared with the control group (MFI=164).</p><p><b>CONCLUSION</b>803G>C mutation of the ABO gene probably underlies the CisAB01 subtype. Fluorescence intensity of A antigens in CisAB01 subtype cases is slightly lower than the normal type, while the B antigen was significantly lower.</p>

Long noncoding RNA MALAT1 regulates Rac1b expression and is associ-ated with colorectal cancer metastasis / 中国病理生理杂志

AIM:ToinvestigatetheroleofMALAT1incolorectalcancermetastasis.METHODS:ThemRNA expression levels of MALAT1 and Rac1b in the tumor and adjacent normal tissues were examined by real-time PCR. MALAT1 was knocked down by siRNA in colorectal cancer cell lines .The expression of Rac1b and the epithelial-mesen-chymal transition markers was examined by Western blot .Cell proliferation was determined by EdU analysis .The effects of MALAT1 on the cell migration and invasion were examined by Transwell assay .RESULTS: The expression of MALAT1 was down-regulated in colorectal cancer .Down-regulation of MALAT1 induced Rac1b overexpression, which in turn in-creased the expression levels of E-cadherin and β-catenin.Furthermore, down-regulation of MALAT1 promoted the cell proliferation, invasion and migration.CONCLUSION:MALAT1 is associated with metastasis of colorectal cancer through regulating the expression of Rac1b and the downstream factors.

Ventilatory efficiency and exercise capacity during exercise in patients with idiopathic pulmonary fibrosis / 中华全科医师杂志

Objective To explore the characteristics of ventilatory efficiency and exercise capacity during cardiopulmonary exercise testing in patients with idiopathic pulmonary fibrosis (IPF).Methods Pulmonary function test,arterial blood gas analysis and cardiopulmonary exercise testing were performed in 28 IPF patients (IPF group) from April 2012 to April 2013 and 28 healthy volunteers (control group).And the relevant parameters were measured and compared.Results No significant differences existed in age [(57.8 ±9.8) vs.(59.2 ±5.5) years],gender or body mass index (BMI) [(23.8 ±2.7) vs.(25.0 ± 2.8) kg/m2,P ＞ 0.05].The paramneters of pulmonary function test,such as forced vital capacity ％ predicted (74.8 ± 14.6 vs.101.8 ± 10.8),forced expiratory volume in 1 second ％ predicted (73.8 ± 14.6 vs.97.0 ± 10.1),maximum voluntary ventilation ％ predicted (77.5 ± 14.9 vs.95.4 ±24.5),total lung capacity ％ predicted (75.6 ± 12.4 vs.99.8 ± 5.4),residual volume ％ predicted (80.7 ± 15.4 vs.95.8 ± 11.3),diffusing capacity of lung for carbon monoxide ％ predicted (66.2 ± 13.7 vs.103.2 ± 17.3) in the IPF group,were significantly lower than those of the control group (P ＜ 0.01).The parameters of arterial blood gas analysis,such as PaO2 [(72.7 ± 7.3) vs.(92.6 ± 3.8) mmHg] and SaO2 (94.3 ± 2.1 vs.98.3 ± 0.7),were lower than those of the control group (P ＜ 0.01).Thus P(A-a) O2 in the IPF group was higher than that in the control group (33.3 ± 5.7 vs.17.8 ± 1.9,P ＜0.01).These results strongly suggested that IPF group had restrictive ventilatory dysfunction and impaired gas exchange.The IPF patients had higher VE/VCO2-slope (37.4 ± 5.3 vs.25.7 ± 2.5,P ＜ 0.01) and lowest VE/VCO2 (39.2 ±6.7 vs.30.6 ± 2.7,P ＜ 0.01) than the controls; VE/VCO2 and VD/VT during every period were significantly higher in the IPF group than those in the control group (P ＜ 0.01) ; during peak exercise,peakLoad％pred (70.4 ±±29.9 vs.104.8 ±29.7,P ＜0.01) and peakVO2％pred (68.7 ±29.8 vs.98.7 ±36.4,P =0.001) were significantly lower in the IPF group than those in the control group.In the IPF group,VE/VCO2@AT,VE/VCO2-slope and lowest VE/VCO2 had a negative correlation with DLCO％pred (r=-0.589,P ＜0.01; r=-0.481,P＜0.05; r=-0.527,P＜0.05).In the IPF group,VE/VCO2@AT,VE/VCO2-slope and lowest VE/VCO2 had a negative correlation with peakVO2％ pred (r =-0.548,P ＜ 0.05 ; r =-0.539,P ＜ 0.05 ; r =-0.564,P ＜ 0.05).So the exercise tolerance and ventilation efficiency of the IPF group decreased significantly.Conclusion Cardiopulmonary exercise testing reveals that the ventilation efficiency of IPF patients decreases significantly so as to seriously affect their exercise tolerance

Therapeutic Effects of Cyclovirobuxine D on Rats with Heart Failure / 医药导报

Objective To exPlore the theraPeutic effect of cycloVirobuxine_D ( CVB_D) on heart failure by obserVing the influences of CVB_D on cardiac function,myocardial morPhology and structure in rats with heart failure. Methods The rat model of heart failure was established by ligating left anterior descending coronary artery,and then the rats were randomly diVided into 5 grouPs,Plus the sham grouP. EchocardiograPhy was used to assess cardiac function,including EF,LVSF,SV,HR,CO and so on. Myocardial tissue section was obtained. Change in the myocardial tissue was obserVed by HE staining. Myocardial ultra_structure was obserVed by using transmission electron microscoPe. Results As comPared with the model grouP,EF and LVSF were significantly increased in medium and high dose CVB_D grouPs (P<0. 05 or P<0. 01),SV was increased in low and medium dose CVB_D grouPs (P<0. 05),CO was increased in low,medium and high dose CVB_D grouPs (P<0. 05),LVIDs was decreased and LVPWs was increased in high dose CVB_D grouP (P<0. 05). Under light microscoPe,changes of myocardial structures were obserVed in different CVB_D dose grouPs: myocardial fiber gradually became tidy, striPes became clear, and contour became clear. The changes of myocardial ultra_structure were obserVed under transmission electron microscoPe: the myocardial fiber breakage was reduced and the number of mitochondria was increased,swelling degeneration was alleViated. Conclusion CVB_D significantly imProVed cardiac contraction and blood PumPing function of rats with heart failure, exerted mitigatiVe effect on damage of cardiac muscle fiber and mitochondria. It suggests that CVB_D may be effectiVe in the treatment of heart failure.

Determination of Camphol and Isoborneol in Shaoshang Yuhe Gao / 医药导报

Objective To establish a capillary gas chromatography method for determination of camphol and isoborneol in Shaoshang yuhe gao ( burn healing cream) . Methods The capillary gas chromatography was adopted under the following conditions: use PEG-2000 as the stationary liquid,nitrogen as carrier gas,ZB-WAX (30 m×0. 25 mm,0. 25 μm) as the chromatographic column,and the flame ionization detector. The column temperature was programmed at 80 ℃ for 5 min as the initial temperature,then raised to 180 ℃ at the rate of 5℃·min-1 and kept for 10 min. The shunt ratio was 101. Results The liner range for camphol was 0. 487 5-31. 25 μg ( r =0. 999 6),and the average recovery was 95. 95%( n =6). The liner range for isoborneol was 0. 487 5-31. 25 μg( r =0. 999 7),and the average recovery was 96. 44%( n =6). Conclusion The method is accurate,sensitive,and can be applied to quality control of shaoshang yuhe gao.

Molecular basis and clinical blood transfusion for cases with ABO typing discrepancy / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the molecular genetic basis of samples with ABO typing discrepancy and provide the guidline for identification and clinical transfusion for these samples.</p><p><b>METHODS</b>Six cases with similar serological characteristics were collected. Serological method, PCR-SSP and direct sequencing of ABO gene were used to explore the underlying mechanism. Condition of clinical transfusion of patients was also reviewed.</p><p><b>RESULTS</b>Three conditions were related with the ABO blood type discrepancy, which included weaken antigen (2 cases), weakened antibody (3 cases) and ABO subtype (1 case). The satisfactory effect of transfusion was achieved in all patients with the principle of the same blood type or the compatible crossmatch.</p><p><b>CONCLUSION</b>Heterogeneity has existed with the ABO group. Indivianals with same reaction pattern may result in different mechanisms.</p>

Application of Tissue-explant Technique in Culturing Rat Submandibular Gland Cells / 中国医科大学学报

Objective To explore the application of tissue-explant technique in culturing rat submandibular gland cell(RSGC)and the characteristics of RSGC.Methods RSGCs were cultured using tissue-explant technique.The cells were purified by enzymatic digestion and differential adhesion.The cell phenotype was identified by cytokeratin-8 immunohistochemical staining.Cellular morphology was observed and photographed under inverted microscope.The cell viability and growth were determined by a double-staining procedure using FDA-PI and MTT assay,respectively.Results Cytokeratin-8 was positive stained in the immunohistochemical staining.The cell viability was more than 95%.The cell growth curve showed that RSGCs were in logarithmic phase since day 5.Conclusion Tissue-explant technique is an easy way to purify plentiful RSGC with normal functions,and it can be used in further research of tissue engineering of submandibular gland.

The levels of serum highsensitive C-reactive protein in different subtypes of stroke and its significance / 中国医师进修杂志

Objective To explore, the levels of serum highsensitive C-reactive protein (hs-CRP) in different subtypes of acute ischemic stroke (AIS) and its significance. Methods According to TOAST criteria, 124 cases of AIS were divided into four groups: large-artery atherosclerosis (LAA,47 cases), small-artery occlusion (SVO,37 cases), cardiogenic cerebral embolism (CE,21 cases), acute stroke of other determined etiology and stroke of other undetermined etiology (ODE and UE, 19 cases). The serum concentration of hs-CRP was measured with the immunoturbidimetry in every patient of each group at 72 hours, 7 and 14 days after admitted to hospital. Meanwhile 81 healthy persons were involved in control group. Results The serum concentrations of hs-CRP at 72 hours, 7 and 14 days after onset were (10.77 ± 4.27),(16.41±5.61), (7.63±3.59) mg/L in LAA group;(3.99± 1.56), (6.45±3.25), (4.22±3.21) mg/L in SVO group; (11.60±4.85), (25.14±7.12), (9.24±4.61) mg/L in CE group; (6.09±2.43), (9.65 ±4.65), (5.89 ± 2.68) mg/L in ODE and UE group. The serum concentrations of hs-CRP in LAA group, CE group, ODE and UE group were significantly higher than those in control group [(4.26 ± 1.34) mg/L] (P <0.05) except for that in SVO group, the level of serum hs-CRP in the three groups showed significant difference among the different time (P< 0.05). Conclusions The level of serum hs-CRP in AIS patients is increased and positively correlated with the disease severity. The changes of hs-CRP level are performed in various subtypes of TOAST, and show time-dependent effects. Inflammatory response may play an important role in the pathological mechanism of AIS.

Genetic diagnosis of four combined deficiency of factor Ⅴ and Ⅷ patients / 中华检验医学杂志

Objective To investigate the genetic diagnosis and molecular pathogenesis of four patients with combined deficiency of coagulation factor Ⅴ and Ⅷ and their family members. Methods The APPT, FT, FⅤ: C, FⅧ: C were detected for phenotypic diagnosis. Thrombin generation assay was applied to determine the generation condition of thrombin in patients and healthy controls. Cenomic DNA was extracted from peripheral blood using the TianGen RelaxCene Blood DNA System;amniotic fluid DNA was extracted with phenol-ethyl ether method. The LMAN1 and MCFD2 genes were analyzed by PCR. Gene mutations were detected with nucleotid sequences by using end-labeling dideoxy method. Results The APTT of Proband 1 was significantly prolonged to 88. 2s and her PT was prolonged to 19. 6 s. The combined deficiency was identified with FⅧ (FⅧ: C 24. 2% ) and FV(FⅤ: C 9. 1% ). Proband 2 and 3 were sisters. The coagulation studies revealed that both of them had prolonged APTT (71.6 s and 74.6 s respectively) and PT (22. 1 s and 18. 3 s respectively). The combined deficiency of FⅤ (FⅤ: C 7. 6% and 14. 5% respectively) and FⅧ( FⅧ: C 25% and 19.6% respectively) were identified. Proband 4 was detected to have the prolonged APTT (70.3 s),PT (18.2 s) and the deficiency of FⅤ(FⅤ: C 9. 4% ) and FⅧ (15. 7% ). The remaining phenotype indicators test of the 4 probands were normal. The diagnosis for the 4 probands was combined deficiency of factor Ⅴ and Ⅷ. The proband 1 was detected to have compound heterozygous mutations in LMAN1 gene while having the LMAN1 and MCFD2 direct gene sequencing. One mutation was a small insertion located on exon 8 [ nt912insA (X71661. 1)] that resulted in p. 305frameshiftX20 and her mother was detected to have the same heterozygous mutation on the the locus. The other mutation was located on exon 11: nt1366C > CT ( X71661. 1 ) , p. 456Arg > Stop which was inherited from her father. Amniocyte DNA was detected to have only one heterozygous mutaion [nt1366C > CT (X71661. 1) , 456Arg > Stop] inherited from the father. No mutation in MCFD2 gene was found in proband 1 and her parents. The analysis of the MCFD2 gene in proband 2 and 3 revealed a novel homozygous single base substitution (nt411T>C) in exon 4, which results in the exchange of the amino acid isoleucine by the amino acid threonine at amino acid position 136 (p. Ile136Thr). Sequencing of the whole LMAN1 gene showed that the proband 4 had one homozygous nonsence mutation in the exon 5 of the LMAN1 ( nt615C >T,p. 202 Arg> Stop). All of the 4 probands with combined deficiency of FⅤ and FⅧ showed declined endogenous thrombin potential in the thrombin generation tests. Conclusion The combined deficiency of FⅤ and FⅧ in the proband 1 results from the compound heterozygous mutations ( nt1366C > CT and nt912insA) in LMAN1 gene, which are inherited from her parents respectively. The prenatal genetic investigation for the patient mother with preganency indicates that the fetus is a female carrier with one mutation (nt1366C > CT) inherited from the father. The homozygous missence mutation ( nt411T > C, p. Ile136Thr) in the MCFD2 gene accounts for the proband 2 and 3. The daughter of the proband 2 is a carrier with a heterozygous mutation inherited from her mother. The homozygous nonsence mutation in the LMAN1 gene of the proband 4 results in the deficency of F Ⅴ and FⅧ.

Study on the molecular mechanism of the inherited factor X deficiency in three unrelated families / 中华检验医学杂志

Objective To identify the clinical features, the molecular diagnosis and the molecular mechanism of three unrelated factor X deficiency families. Methods Three probands were male and the diagnosis was validated by coagulant parameters. The F X coagulation activity ( F X∶ C ) and antigen (FX∶ Ag) were tested by clotting test and ELISA method. The cross-corrected test was used to rule out the inhibitor of FX in plasma. Thrombin generation test was evaluated. The antigen and the molecule weight of the FX in plasma were measured with western blotting. Gene mutations were analyzed in the probands and their family members with PCR and DNA sequencing. FX expression plasmids were constructed and transientby being transfected into 293T cells. FX: C and FX: Ag of the expression products were tested. Results APTT and PT in proband 1 were obviously prolonged, 113.4 s and 62.3 s, respectively. And there was no inhibitor in plasma. The thrombin generation was lower compared to normal reference. APTT and PT in proband 2 were 56. 5 s and 28.7 s. There was no inhibitor in the plasma. The thrombin generation was 1 101.5 nmol · min. APTT and PT in proband 3 were 117.3 s and 44. 3 s. The thrombin generation was 782.5 nmol · min. FX∶ C and FX∶ Ag in proband 1 were 1.4% and 3.6%, with a homozygous mutation in FX gene (Ser425→Pro). In vitro expression of the mutation showed a normal synthesis in the cell but secretion dysfuntion. In proband 2 F X: C and F X: Ag were 2. 2% and 5. 5%, with two heterozygous mutations in FX gene (Ala-29→Pro and Phe324→Leu). The Ala-29 → Pro mutation led to significantly reduced expressions of FX in both cell lysate and cell culture supernatants compared to wild-type plasmid,(41.32 ±5.21 )% and(6. 30 ± 1.84)% respectively. However Phe324→Leu mutation almost did not affect the FX synthesis. FX: C and FX: Ag in proband 3 were 2. 2% and 35%, with two heterozygous mutations in FX gene( Ala235→Thr and Arg347→Cys). The expressions of these two mutant FX proteins in cell lysate were similar to those of wild-type but obviously lower in the supernatant. Conclusions Five mutations of F X gene are found in this study. These mutations (Ser425Pro, Phe324Leu, Ala235Thr and Arg347Cys)can not affect F X protein synthesis. However Ala-29Pro mutation can reduce F X protein synthesis and cause secretion dysfunction.

Effect of jejunal casein perfusion on pancreatic exocrine secretion in experimental acute necrotic panereatitis rats / 中华胰腺病杂志

Objective To investigate the effect of jejunal casein perfusion on pancreatic exocrine secretion in experimental acute necrotic pancreatitis (ANP) rats and analyze the neuromechanism that may be involved. Methods 30 SD rats were randomly divided into three groups (control group, ANP group and ANP jejunal nutrition group). Experimental ANP was induced by intra pancreatic duct injection of sodium taurocholate (STC). Animals in ANP jejunal nutrition group were given jejunal casein perfusion 24h after model induction, while control group and ANP group received jejunal saline perfusion. Pancreatic juice was collected every 15 min for six times and the volume of pancreatic juice and protein in pancreatic juice were detected. After jejunal nutrition c-Fos expression in nucleus tractus solitarii (NTS) was determined by immunohistochemistry method in three groups. Results There was no significant difference between the volume of pancreatic juice at different time points in ANP group and ANP jejunal nutrition group, however, these parameters were significantly lower than that of control group (P < 0. 05). There was no significant difference among the 3 groups in the protein level in the pancreatic juice during jejunal nutrition infusion, however, during the periods of 0 ～ 15 min, 15 ～30 min, 30 ～45 min and 75 ～90 min, the protein levels in the pancreatic juice in ANP and ANP jejunal nutrition group were lower than that of control group (P < 0.05). After jejunal perfusion, c-Fos expression was found in ANP jejunal nutrition group but not found in ANP and control groups. Conclusions Jejunal casein perfusion enhanced NTS c-Fos expression, but did not increase the volume of pancreatic juice and protein.

Effects of homocysteine on expression of TNFSF4 gene and interleukin-10 in monocytes cultured in vitro / 中国组织工程研究

BACKGROUND: Inflammatory response is one of potential cellular mechanisms by which hyperhomocysteinemia accelerates atherosclerosis. There are still many details need to be illuminated. TNFSF4 gene and interleukin (IL)-10 are two inflammatory factors related to atherosclerosis. But their correlation with hyperhomocysteinemia is seldom reported. OBJECTIVE: To investigate the effects of homocysteine (Hcy) on expression of TNFSF4 gene and IL-10 in the monocytes cultured in vitro. DESIGN, TIME AND SETTING: A randomized control experiment in vitro was conducted from July to December in 2007 in the Pathophysiology Laboratory of West China School of Preclinical and Forensic Medical Sciences of Sichuan University. MATERIALS: THP-1 monocytes were offered by the Pathophysiology Laboratory of West China School of Preclinical and Forensic Medical Sciences of Sichuan University); Hcy, dexamethasone (Dex) and vitamin D3 (Vit D3) were all purchased from Sigma. METHODS: Cultured THP-1 monocytes were induced to macrophages by 0.1 ?mol/L phorbol myristate acetate, and the differentiated THP-1 macrophages were incubated with 200 ?mol/L Hcy for 24, 48 and 72 hours respectively. Then cell supernatant and lysate were collected as condition medium. Any other differentiated THP-1 macrophages were incubated with a combination of Dex and Vit D3 for 6 days, and then the condition mediums were added to incubate together for 24 hours. Cells were divided into blank control group, 200 ?mol/L Hcy group, Dex+Vit D3 group, 24-hour Hcy+Dex+Vit D3 group, 48-hour Hcy+Dex+Vit D3 group, and 72-hour Hcy+ Dex+Vit D3 group. MAIN OUTCOME MEASURES: The expression level of TNFSF4 mRNA was determined by reverse transcription polymerase chain reaction. The expression level of IL-10 protein was determined by ELISA. RESULTS: Compared with blank control group, no significant difference of TNFSF4 mRNA expression was found in the 24-hour Hcy group, while the expression level was significantly higher in 48-hour and 72-hour Hcy groups. Compared with Dex+Vit D3 group, the IL-10 expression of all Hcy+Dex+Vit D3 groups was significantly lower than that in the Dex+Vit D3 group. CONCLUSION: Hcy may increase TNFSF4 mRNA expression and decrease IL-10 expression in THP-1 macrophages.

Effects of fructose-1,6-diphosphate of low and high dosages on islet cells damaged by interleukin-1 beta / 中国组织工程研究

BACKGROUND: Fructose-1,6-diphosphate (FDP) of certain dosage plays a protective role in the pancreatic islets damaged by interleukin-1 beta (IL-1β), and there are different effects of FDP of low and high dosages.OBJECTIVE: To investigate the effects of FDP of low and high dosages on the islet cells damaged by IL-1β.DESIGN: A grouped design and controlled animal experiment.SETTING: Department of Physiology, North Sichuan Medical College.MATERIALS: The experiments were carried out in the Tumor Laboratory and Central Laboratory of Rheumatology and Immunology, Department of Surgery, North Sichuan Medical College between July 2004 and February 2006. Twenty Wistar rats within 1-3 days after birth were selected.METHODS: The pancreases of the rats were removed to collect islet cells, and then the cells were divided into normal control group, IL-1β damaged group, IL-1β+1, 25, 50 mmol/L FDP groups. The cellular activity was detected with methyl-thiazol-tetrazolium (MTT) assay, basic amount of insulin secretion and that stimulated by high glucose with radioimmunoassay, content of nitric oxide and activity of nitric oxide synthase (NOS) with nitric oxide and NOS kits, and the with [Ca2+]i with Fura-2fluorescent assay.MAIN OUTCOME MEASURES: Activity of islet cells; basic amount of insulin secretion and that stimulated by high glucose; content of nitric oxide and activity of NOS; [Ca2+]i.RESULTS: ① The activities (A values) of the islet cells in the IL-1β damaged group, IL-1β+1, 25, 50 mmol/L FDP groups were obviously lower than that in the normal control group (0.116±0.012, 0.129±0.008, 0.125±0.015, 0.120±0.016, 0.252±0.020, P ＜ 0.01). The activities (A values) of the islet cellswere not significantly different from that in the IL-1β damaged group (P ＞ 0.05) when the FDP dosage was too low (1 mmol/L) or too high (25 mmol/L). ② The basic amount of insulin secretion and that stimulated by glucose were significantly lower in the IL-1β damaged group, IL-1β+1, 25, 50 mmol/L FDP groups than in the normal control group [(237.00±22.21), (230.83±11.58), (225.16±12.46), (220.50±15.63),(425.67 ±16.85) mIU/L; (90.17 ±6.11), (96.62 ±8.64), (87.66-±8.24),(85.46±9.59), (204.50±10.78) mIU/L, P ＜ 0.01], and there were no significant differences between the FDP groups of Iow and high dosages and the IL-1β damaged group (P ＞ 0.05). ③ The NOS activity and content of nitric oxide in the supernatant were obviously higher in the IL-1β damaged group than in the normal control group [(332.07±25.34), (144.86±12.17) μkat/L;(457.64±19.29), (84.67±10.23) μmol/L, P ＜ 0.01], and those in the IL-1β+1, 25, 50 mmol/L FDP groups were not significantly different from those in the IL-1β damaged group. ④ The [Ca2+]i concentration in islet cells was obviously higher in the IL-1β damaged group than in the norrmal control group [(328.50±26.28), (73.42±1.79) nmol/L, P ＜ 0.01], but obviously lower in the IL-1β+1, 25, 50 mmol/L FDP groups than in the IL-1β damaged group [(152.72± 11.86), (216.39±15.32), (233.61±21.76),(328.50±26.28) nmol/L, P ＜ 0.01].CONCLUSION: FDP of low and high dosages can not protect the islet cells damaged by IL-1β.