High-Throughput Sequencing Technology for Detection of Gene Mutations in Myeloid Malignancies and Its Clinical Prognostic Significance / 中国实验血液学杂志

OBJECTIVE@#To detect the gene mutations in patients with myeloid malignancies by high-throughput sequencing and explore the correlation between gene mutations and prognosis.@*METHODS@#A retrospective analysis was performed on 56 patients with myeloid malignancies who were hospitalized in the department of hematology, Peking University International Hospital from January 2020 to May 2021. The genetic mutations of the patients were detected by next-generation sequencing technology, and the correlation between the genetic mutations and prognosis of myeloid malignancies was analyzed.@*RESULTS@#In 56 patients, the number of mutated genes detected in a single patient is 0-9, with a median of 3. Sequencing results showed that the most common mutated genes were RUNX1(21.4%), TET2(17.9%), DNMT3A(17.9%), TP53(14.3%) and ASXL1(14.3%), among which the most common mutations occurred in the signaling pathway-related genes (23.3%) and the transcription factor genes (18.3%). 84% of the patients carried multiple mutated genes (≥2), and correlation analysis showed there were obvious co-occurring mutations between WT1 and FLT3, NPM1 and FLT3-ITD, and MYC and FLT3. TP53 mutation was more common in MDS patients.The overall survival time of patients with NRAS mutation was significantly shortened (P =0.049). The prognosis of patients with TP53 mutation was poor compared with those without TP53 mutation, but the difference wasn't statistically significant (P =0.08).@*CONCLUSION@#The application of next-generation sequencing technology is of great significance in myeloid malignancies, which is helpful to better understand the pathogenesis of the disease, to judge the prognosis and to find possible therapeutic targets.

Clinical characteristics and prognosis of MLL-AF6 positive patients with acute myeloid leukemia / 北京大学学报(医学版)

OBJECTIVE@#To investigate the clinical features and prognosis of acute myeloid leukemia (AML) patients with the mixed lineage leukemia (MLL) gene rearrangements AF6 (MLL-AF6) positive.@*METHODS@#In the study, 11 patients who were newly diagnosed with MLL-AF6 positive AML were analyzed retrospectively, related literature was reviewed to clarify the clinical features and prognosis of MLL-AF6 positive patients.@*RESULTS@#Among the 11 patients, there were 6 males and 5 females, with a median age of 36 years. Six patients were diagnosed with AML M5 and five with M4 according to FAB classification (French-American-British classification systems). Gingival swelling and pain occurred in 6 cases and fever occurred in 5 cases. At first diagnosis, the median white blood cells were 55.5×109/L. Immunotype showed the expression of myeloid/monocyte and early stem cell series antigens. The expression level of MLL-AF6 fusion gene (real-time quantitative PCR) was 14.2%-214.5%, and 6/11 cases (54.5%) were associated with high EVI1 gene expression. Mutations of KRAS, TET2, ASXL1, TP53, DNMT3A, and FLT3-ITD were detected by next generation sequencing (NGS) in 4 patients. Chromosome G banding examination showed that 2 cases were t(6;11)(q27, q23) with complex karyotype abnormality, 4 cases with +8 abnormality and 2 cases with normal karyotype. Hematological complete remission (CR) was achieved in 8/11 patients (72.7%) after conventional induction chemotherapy, and primary drug resistance was observed in 3 patients. Two of the eight patients with CR were negative for minimal residual disease (MRD), with a median CR duration of 4.5 months. Two patients with positive MRD and three patients with refractory recurrence underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT), but all died due to leukemia progression. At the end of follow-up on December 1, 2019, 2 patients were alive and 9 died, with median survival time of 9 months.@*CONCLUSION@#The AML patients with MLL-AF6 positive were mostly young, the majority of FAB types were M4 and M5, and most of the patients often had fever as the first symptom, with increased white blood cells, accompanied by organ infiltration, and high EVI1 gene expression. The hematological remission rate of routine chemotherapy is not low, but it is difficult to achieve molecular remission, most of which have early recurrence. Early allo-HSCT in a molecular negative state may prolong the CR duration.

Comparison of the Tumor-forming Rate between the SCID and NOD/SCID Mice used to Set up Acute Myeloid Leukemia Model / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To compare the tumor-forming rate between the SCID and NOD/SCID mice to set up acute myeloid leukemia (AML) mouse model.</p><p><b>METHODS</b>The SCID and NOD/SCID mice were injected with HL-60 cells in to the abdominal cavity in order to contruct the AML mouse model. The gereral status of mice was observed, the positive rate of CD33 in bone marrow cells was detected by flow cytometry, the mouse model was identified by pathologic examination.</p><p><b>RESULTS</b>The tumor-forming rate in constructed model using SCID mouse was 30%, while the tumor-forming rate in constructed model using NOD/SCID mouse was 100%.</p><p><b>CONCLUSION</b>Compared with SCID mice, the tumor-forming rate in NOD/SCID mice injected with HL-60 cells is high, the incidence of AML is stable, suggesting that the NOD/SCID mouse model is more suitable to explore the pathogenesis of leukemia.</p>

miRNA-101 inhibits the expression of the enhancer of zeste homolog 2 in androgen-independent prostate cancer LNCaP cell line / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.</p><p><b>METHODS</b>We divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.</p><p><b>RESULTS</b>Green fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.</p><p><b>CONCLUSION</b>miRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.</p>

Construction of a recombinant BCG secreting BP26 and the effects of BP26 on CD4+ and CD8+ T cells in mice / 中国地方病学杂志

Objective To develop a BP26 recombinant BCG (rBCG-BP26) vaccine,and to observe the effects of rBCG-BP26 on CD4+,CD8+ T cells in immunized mice.Methods The recombinant shuttle vector pMV261-Ag85B-BP26 was constructed by using traditional molecular biological technology.The recombinant strains were obtained by kanamycin resistance screening and PCR identification after electroporation.Western blotting was used to detect the expression of recombinant BP26 vaccine in immunized mice.Safety experiment was carried out in three different groups:the target experiment(rBCG-BP26) group,the positive control(BCG) group and the negative control(PBS) group,15 BALB/c mice in each group.Intradermal inoculations of 100 μl rBCG-BP26 [containing 106 colony forming units(CFU)],BCG,and PBS were carried out,respectively.Signs of mice in each group were observed.After immunization for 10,20,30,and 40 days,body weight was weighed,and tail blood was collected to observe the change of peripheral blood CD4+ and CD8+ T cells by flow cytometry.Results The rBCG-BP26 was successfully constructed.The expression of BP26 protein was detected in the liquid medium and the bacteria cells.The results of safety test analysis showed that there were no significant differences in signs and body weights(F=2.468,0.331,1.520,0.739,all P＞ 0.05),between PBS group[ (19.24 ± 0.54),(21.37 ± 0.66),(22.83 ± 0.62),(25.06 ± 0.37)g],BCG group[ (19.90 ± 0.02),(21.53 ± 1.57),(21.95 ± 0.55),(24.70 ± 0.39)g]and rBCG-BP26 group[ (19.16 ± 0.55 ),(20.89 ± 0.20),(22.15 ± 0.76),(24.60 ± 0.64)g].The results of flow cytometry showed that the percentages of CD4+ T cell level were lower in BCG group(26.70％,33.07％) and rBCG-BP26 group( 13.40％,26.70％) than that of the PBS group(33.85％,29.33％) and the values of CD4+/CD8+ T cells increased in rBCG-BP26 group (0.69％,1.27％,1.57％,1.70％ ) 10,20 and 30 days after immunization.Conclusions Recombinant BCG-BP26 vaccine strain can express brucella BP26 protein efficiently.Furthermore,its virulence is mild,and it can activate CD4+,CD8+ T cells in the body.It can be used as one of candidate vaccine strain against brucellosis.

Effects of stimulating in the cavum concha on glucometabolism in patients with diabetes / 中国针灸

<p><b>OBJECTIVE</b>To explore the intervention effect of the auricular stimulator in the cavum concha for diabetes.</p><p><b>METHODS</b>Forty-five cases were treated with auricular simulator in the cavum concha for 30 min, once daily for consecutive 3 months. The changes of the fasting plasma glucose (FBG), blood glucose load after 2-hour 75 g oral glucose tolerance test (P2 BG)and glycosylated hemoglobin (HbA1c) were compared before and after treatment.</p><p><b>RESULTS</b>The level of the HbA1c was obviously decreased (P < 0.05, P < 0.01), and there were also statistically significant differences in FBG and P2 BG after the treatment (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>With the auricular stimulator, the stimulation in the cavum concha is benefit for the improvement of HbA1c of the diabetes.</p>

Effect of Lianggesan on the expression of signal transducer and activator of transcription 1 in a rat model of lipopolysaccharide-induced acute lung injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Lianggesan on the expression of signal transducer and activator of transcription 1 (STAT1) in rats with lipopolysaccharide (LPS)-induced acute lung injury and explore the possible mechanisms of the therapeutic effects.</p><p><b>METHODS</b>Endotoxemia was induced in Wistar rats by intravenous injection of LPS (5 mg/kg). The rats were randomly divided into 6 groups, namely the control group, acute lung injury group (LPS group), 3 Lianggesan groups treated at different doses, and LPS+DEX treatment group. Each group, except for the control group, was further divided into 5 subgroups and examined at 1, 2, 4, 8 and 16 h after LPS injection. Western blotting was used to detect the protein expression of STAT1 and p-STAT1 in the lung tissue.</p><p><b>RESULTS</b>In LPS group, the expression of STAT1 began to increase at 1 h following LPS injection, reaching the peak level at 4 h; the peak expression of p-STAT1 occurred at 2 h after LPS administration (P<0.01). Compared with LPS group, the 3 Lianggesan groups and DEX group showed significantly decreased expressions of STAT1 and p-STAT1 at 2, 4 and 8 h after LPS injection (P<0.05 or 0.01).</p><p><b>CONCLUSION</b>Abnormal expression of STAT1 occurs in the lung tissue in the event of ALI. Lianggesan can relieve LPS-induced acute lung injury in rats by decreasing the expression of STAT1 and p-STAT1.</p>

Prenatal risk factors for neonatal asphyxia: how risk for each? / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Neonatal asphyxia is the third leading cause of neonatal death and main cause of long-term neurodevelopmental handicap throughout the world. Prevention is more important than treatment. Most previous reports are limited to retrospective investigations of the relationships between some prenatal risk factors and low Apgar scores. This study was designed to prospectively investigate the relationship between prenatal risk factors and neonatal asphyxia and the influence of single or multiple risk factors on the incidence of neonatal asphyxia, and examine significant risk factors for neonatal asphyxia.</p><p><b>METHODS</b>From April 2002 through October 2004, a total of 10 376 live-born newborns were enrolled. Forty-six prenatal risk factors were investigated. Neonatal asphyxia was diagnosed based on the following four items: 1. 1-min Apgar score <or=7; 2. Umbilical artery blood pH<7.20; 3. At least one organ had evidence of asphyxial injury; 4. Other causes of low Apgar score were excluded. The number, the constituent ratio and the exposure frequency of newborns with single or multiple risk factors were counted. The influence of risk factors on the incidence of asphyxia was analyzed. The significant risk factors were screened by single logistic regression analysis and forward stepwise conditional multiple logistic regression analysis, with enrolled threshold alpha<or=0.05, excluded threshold alpha>or=0.10 and p<0.05 as significant. The OR and 95%CI were calculated for each significant risk factor.</p><p><b>RESULTS</b>Of the 10 376 newborns, 8 530 cases (82.21%) had 1-9 risk factors, and asphyxia occurred in 117 cases (1.13%) out of the 8 530 cases. In the 1 846 cases without risk factors, none had asphyxia (x2=25.6, p<0.01). The incidence of asphyxia increased with increasing numbers of risk factors, from 0.23% in newborns with one risk factor to 14.29% in newborns who had nine risk factors (r=0.96, p<0.01). Twelve significant risk factors identified were as follows: ominous fetal heart rate patterns (OR=17.1,95%CI:11.2-25.9), placenta abruption (OR=15.2, 95% CI: 4.5-51.8), maternal lung diseases (OR=11.5, 95% CI:1.4-91.3), fetal acidosis (OR=6.1, 95% CI:1.5-24.1), placenta previa (OR=5.0,95% CI:1.5-16.9), breech delivery (OR=4.5, 95% CI: 2.1-9.9), meconium stained amniotic fluid (OR=3.2, 95% CI:2.2-4.8), forcepsjassisted delivery (OR=3.2, 95%CI: 1.1-9.9), prolonged labor (OR=2.94, 95%CI:1.5-5.8), abnormal utero contraction (OR=2.8, 95% CI:1.7-4.6), and premature delivery (OR=2.5,95%CI:1.4-4.8). Cesarean section had a protective effect (OR=0.6, 95% CI:0.4-0.9) (all p<0.05).</p><p><b>CONCLUSIONS</b>It is very important to prevent perinatal asphyxia by systematically examining prenatal risk factors and giving interventions for the newborns with risk factors, especially those with the above significant risk factors or with multiple risk factors. Proper cesareon section according to indications might be helpful to decrease the incidence of birth asphyxia.</p>

Effects of different oxygen flow rates on myocardial ischemia-reperfusion injury in rabbits / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effects of different oxygen flow rates during cardiopulmonary bypass (CPB) on myocardial ischemia-reperfusion (IR) injury in rabbits.</p><p><b>METHODS</b>Thirty rabbits were randomized equally into groups A, B and C to receive controlled oxygen reperfusion at low, normal and high flow rates (25, 50, and 80 ml.kg(-1).min(-1), respectively). Serum concentration of CK-MB and cTnT were tested by ELISA before the operation (T0) and after 30 min (T1), 2 h (T2), 12 h (T3) and 24 h (T4) of reperfusion. W/D, SOD and MDA of the myocardium were determined before and at 60 min after reperfusion. The ultrastructural alterations of the myocardium were observed.</p><p><b>RESULTS</b>Serum concentration of CK-MB and cTnT in the 3 groups increased significantly after the operation, and their levels were the lowest in group A (P<0.05). W/D and MDA in the myocardium was also the lowest, while SOD the highest in group A (P<0.05). Ultrastructural pathologies were found in all the 3 groups, but relatively mild in group A.</p><p><b>CONCLUSION</b>Low oxygen flow rate during controlled reperfusion may protect the myocardium from IR injury in rabbits.</p>

Application and curative effect of plasma exchange in myasthenia gravis patients with thymectomy / 中南大学学报(医学版)

OBJECTIVE@#To explore the application of plasma exchange in myasthenia gravis (MG) patients with thymectomy, and to evaluate its curative effects.@*METHODS@#The clinical data of 50 patients with thymectomy were analyzed retrospectively, and the patients were divided into Group A (23 cases, undergoing thymectomy and plasma exchange) and Group B (27 cases, undergoing thymectomy). The difference of serum concentration of AChRab was examined by ELISA, and the time of auxiliary breath, crisis of myasthenia and clinical evaluation in the 2 groups were compared.@*RESULTS@#Serum concentration of AChRab in Group A descended obviously after the plasma exchange. One day and one week after the operation, the AChRab concentration in Group A was obviously lower than that in Group B, but there was no difference one month after the operation. One week after the operation, the time of auxiliary breath,crisis of myasthenia and clinical evaluation in Group A were less than those in Group B.@*CONCLUSION@#Plasma exchange together with thymectomy can improve the temporary curative effects in patients with MG. It is safe and effective for MG.

Influence of daidzein on left ventricular remodeling in rats with hypertrophic myocardium induced by pressure overload / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the protective effects of daidzein (DD) on ventricular remodeling in rats with myocardial hypertrophy induced by pressure overload and its mechanism.</p><p><b>METHOD</b>Myocardial hypertrophy model of rats induced by pressure overload was prepared by constricting abdominal aorta. The operated rats were randomly divided into sham operated control group, aorta-constricted model group and three DD groups (30, 60, 120 mg kg(-1)). Four weeks later, the heart-weight (HW), left ventricular weight (LVW), the ratio of HW/BW and LVW/BW (LVI) and the cardio-myocyte diameters (MD) after dyeing by HE colar were measured. The hydroxyroline, nitric oxide (NO) and the activity of nitric oxide synthetase (NOS) and Na+ -K+ -ATPase, Ca2+ -ATPase in left ventricle were quantified with spectrophotometry and the angiotension II (Ang II) in left ventricle and serum was messured with radioimmunoassay.</p><p><b>RESULT</b>After treatment of the left ventricular with DD, vs aorta-contricted model group, NO content, cNOS and Na+ -K+ -ATPase, Ca2+ -ATPase activity were significantly increased, the content of AngII in left ventricle and serum and iNOS activity and the ratio of HW/BW, LVI, MD were significantly reduced.</p><p><b>CONCLUSION</b>DD has protective effects on ventricular remodeling in rats with myocardial hypertrophy induced by pressure overload and its mechanism may be related to raising NO content and reducing the level of Ang II.</p>

Construction and identification of a plasmid with EGFP reporter gene for enhancer function analysis / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a plasmid vector with EGFP reporter gene for functional analysis of enhancers.</p><p><b>METHODS</b>EGFP DNA was amplified by PCR from plasmid pEGFP-N1 DNA and subcloned into plasmid PGL3-promoter backbone without luc(+) gene to construct the enhancer-identifying vector pEGFP-enhancer. Different copies of hypoxia response element (HRE) sequence were synthetized and subcloned into the multiple cloning site of the plasmid pEGFP-enhancer. Using Lipofectamine 2000, the recombined pEGFP-HRE and pEGFP-5HRE plasmids were transfected into the Hela cells respectively. After hypoxic or normoxic cell culture, EGFP expression in the cells was detected by flow cytometry and fluorescence microscopy.</p><p><b>RESULTS</b>After hypoxic exposure, the fluorescence intensity of EGFP in the Hela cells transfected with the plasmid increased with the enhancer HRE copies, while the fluorescence intensity underwent no significant changes after normoxic cell culture.</p><p><b>CONCLUSION</b>we have successfully constructed the enhancer expression vector plasmid pEGFP-enhancer, which can identify the activity of the enhancers through EGFP expression.</p>

Experimental study of the protective effect of astragalus injection(黄芪注射液) on brain after craniocerebral injury / 中国中西医结合急救杂志

Objective To explore the protective effect of astragalus injection(黄芪注射液)on cerebral cells after craniocerebral injury.Methods Sixty male Sprague-Dawley(SD)rats were randomly divided into five groups(each n=12).In the control group,only anesthesia and cranial bone surgery was performed, which did not impact on dura mater.In the model group,a freefall method was used to cause the left parietal lobe of brain limited contusion and saline was given.In the low,medium and high-dose astragalus injection (5,10,20 ml/kg)treatment groups,astragalus injection was injected intraperitoneally after injury at 30 minutes and 12 hours after injury respectively.After 24 hours of injury in each group,the rats were executed.The brain tissue from contusion was taken out and content changes of Ca~(2+),Mg~(2+),glutamate (Glu)and plasma endothelin(ET)were measured.Results In the model group,the Ca~(2+),Glu and ET contents in the rat brain tissues were significantly higher,but Mg~(2+)content was markedly lower than that of the control group(all P0.05).Conclusion Astragalus has a protective effect on nerve cells after craniocerebral injury.

Studies on phenolic compounds from Polygonum aviculane / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from the aerial parts of Polygomun aviculane.</p><p><b>METHOD</b>The chemical constituents were isolated by silica gel column chromatography and preparative silica thin layer chromatography, and their structures were elucidated on the basis of physico-chemical evidences and spectroscopic analysis (IR, MS, 1H and 13C-NMR).</p><p><b>RESULT</b>Seven phenolic compounds were identified as rosmarinic acid (1), gallic acid (2), gentisic acid 5-O-(6'-O-galloyl)-beta-D-glucopyranoside (3), caffeic acid (4), p-coumaric acid (5), ethyl caffeate (6) and acteoside (7), respectively.</p><p><b>CONCLUSION</b>Compounds 1, 3, 6 and 7 were isolated from this plant for the first time. These results provided theoretical evidences for the further bioactive investigation on this plant.</p>

Effect of citicoline on spatial learning and memory of rats after focal cerebral ischemia / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of citicoline on spatial learning and memory of rats after focal cerebral ischemia.</p><p><b>METHODS</b>The rats were randomly divided into sham-operation group, ischemia control group and citicoline group. In the later two groups, focal cerebral ischemia model was established by introducing an intraluminal filament into the left middle cerebral artery, and citicoline (500 mg/kg) or 0.9% NaCl was administered intraperitoneally once a day for 2 weeks after the operation. The rats in the sham-operation group were not subjected to middle cerebral artery occlusion (MCAO) with intraluminal filament. The spatial learning and memory functions of the rats were evaluated by Morris water maze test 15 days after MCAO for 5 days.</p><p><b>RESULTS</b>The rats in ischemia control group exhibited serious spatial learning and memory deficits in both place navigation test and spatial probe test. In the former test, the mean escape latency of citicoline-treated rats were significantly shorter than that of ischemia control rats (P<0.01), and in the latter test significant diffidence was noted between citicoline and ischemia control groups in the percentage time spent in the former platform quadrant and frequency of crossing the former platform (P<0.05).</p><p><b>CONCLUSION</b>Citicoline can improve the spatial learning and memory function of rats after focal cerebral ischemia.</p>

Cloning, expression, and antibody preparation of nestin with immunohistochemical analysis / 南方医科大学学报

<p><b>OBJECTIVE</b>To obtain recombinant nestin and prepare anti-nestin polyclonal antibody (mAb) to explore the biological roles of nestin in the central nervous system development.</p><p><b>METHODS</b>The nestin cDNA was cloned from human neural stem cells by RT-PCR and ligated to prokaryotic expression plasmid pQE30 for construction of the recombinant vector pQE30-nestin. After sequencing, the recombinant vector was transformed into E.coli M15 and His-tagged nestin was induced by IPTG. The nestin was purified by Ni-NTA affinity chromatography column and characterized by SDS-PAGE and Western blotting. BALB/c mice were immunized with the purified recombinant protein to prepare the antiserum, which was analyzed by Western blotting, enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry.</p><p><b>RESULTS</b>The nestin gene was successfully cloned from human neural stem cells, which was identical to that reported in GenBank. After IPTG induction, the E.coli transformed with pQE30-nestin plasmid expressed a 25,000 His-tagged protein, which was successfully purified and identified as nestin by Western blotting. Western blotting, ELISA and immunohistochemistry demonstrated that the antiserum could specifically bind to the recombinant nestin as well as to nestin in fetal human and rat brains.</p><p><b>CONCLUSION</b>We successfully cloned the nestin gene and expressed the nestin, and nestin mAb prepared can specifically recognize not only the recombinant nestin, but also nestin from human and rats brain tissues.</p>

Effect of histamine on intracortical blood vessels of rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate histamine-induced changes of the intracortical vessels in the cortical slice of rat brain.</p><p><b>METHODS</b>Immunohistochemistry was employed to detect the expression of H1 and H2 receptors in the intracortical blood vessels of rats. Histamine-induced constriction of the intracortical blood vessels of the brain slices was observed with differential interference contrast microscope. Measurements of the luminal diameter were made on-line during the course of the experiment and confirmed off-line from the stored images. In order to observe whether histamine H1 and H2 receptors affected histamine-induced constriction, the intracortical blood vessels in the brain slices were pre-treated with H1 receptor antagonist diphenhydramine and H2 receptor antagonist cimetidine.</p><p><b>RESULTS</b>Expression of H1 and H2 receptors was detected in the intracortical blood vessels of the rat brain. Histamine (1-100 micromol/L) induced a concentration-dependent constriction from (1.48-/+0.67)% to (32.91-/+7.91)%. The reactions to each histamine concentration were significantly (P<0.01) different from each other, with the exception of the highest histamine concentrations (30 and 100 micromol/L) when maximal constriction due to histamine were observed (P>0.05). With pre-treatment of the slice with 10 micromol/L diphenhydramine, application of histamine did not elicit constriction. Pre-treatment of the slice with 10 micromol/L cimetidine did not completely inhibit but somehow significantly weakened vascular constriction in response to histamine treatment at 10 and 30 micromol/L (P<0.05).</p><p><b>CONCLUSION</b>Histamine can induce constriction of the intracortical blood vessels, which is mediated by H1 receptor.</p>

Protective effects of ulinastatin on the lung injury during cardiopulmonary bypass / 中南大学学报(医学版)

OBJECTIVE@#To determine the protective effects and mechanism of ulinastatin on the lung injury during cardiopulmonary bypass (CPB).@*METHODS@#Thirty patients with rheumatic heart disease (RHD) were divided into 2 groups randomly. The ulinastatin group (Group U, n = 15) received 1 x 10(4)U/kg ulinastatin intravenously before the CPB and the same amount of ulinastatin was added into the primary solution. The control group (Group C,n = 15) received normal saline instead of ulinastatin. A brochioalveolar lavage was performed at 2 h after the cardiopulmonary bypass. Polymorphonuclear neutrophil elastase, tumor necrosis factor-alpha and MDA contents in the brochioalveolar lavage fluids were measured, and the lung oxygenate index was measured preoperatively and at 1 and 4 h after CPB termination.@*RESULTS@#Polymorphonuclear neutrophil elastase, tumor necrosis factor-alpha and MDA contents of Group U in the brochioalveolar lavage fulids were significantly lower than those of Group C (P < 0.05), and the lung oxygenate index of Group U at 1 and 4 h after CPB termination was also significantly lower than that of Group C. A significant increase of lung oxygenate index occurred in both groups at 1 and 4 h after CPB when compared with the same group at the baseline before CPB (P < 0.05).@*CONCLUSION@#Ulinastatin has the protective effects on the lung injury during CPB by decreasing polymorphonuclear neutrophil elastase, alleviating lung inflammatory reaction and reducing oxygen free radicals.

Tea saponins affect in vitro fermentation and methanogenesis in faunated and defaunated rumen fluid / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The effect of tea saponins (TS) on rumen fermentation and methane emission was examined using an in vitro gas production technique named Reading Pressure Technique. Three levels of TS addition (0, 0.2, 0.4 mg/ml) were evaluated in the faunated and defaunated rumen fluid. Compared to the control, TS addition decreased the 24 h gas production in the faunated rumen fluid, but had a minor effect on gas yield in the defaunated rumen fluid. The TS significantly reduced methane production in vitro. In the faunated rumen fluid, 0.2 or 0.4 mg/ml TS decreased the 24 h methane emission by 12.7% or 14.0%, respectively. Rumen fluid pH value was affected neither by TS addition nor by defaunation. The TS addition had only minor effects on volatile fatty acids, but the yield and pattern of volatile fatty acids were greatly affected by defaunation. While the molar proportion of acetate was not affected by defaunation, the propionate was significantly increased and the butyrate significantly decreased. Ammonia-N concentration and microbial protein yield were influenced by TS inclusion and defaunation. Inclusion of 0.4 mg/ml TS increased the microbial protein mass by 18.4% and 13.8% and decreased the ammonia-N concentration by 8.3% and 19.6% in the faunated and defaunated rumen fluid, respectively. Protozoa counts were significantly reduced by TS inclusion. The current study demonstrated the beneficial effect of TS on methane production and rumen fermentation, and indicated that this may be due to the effect of the associated depression on protozoa counts.

Effects of supplementary urea-minerals lick block on the kinetics of fibre digestion, nutrient digestibility and nitrogen utilization of low quality roughages / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Three yearling lambs with a rumen cannula were used to investigate the effects of supplementation with an urea-minerals lick block (ULB) on the kinetics of ruminal fibre digestion, nutrient digestibility and nitrogen (N) utilization of rice straw (RS), ammonia bicarbonate (AB)treated RS (ABRS) and hay prepared from natural pasture. The digestibility of dry matter and organic matter of RS increased by 13.1% and 12.7% (P<0.05) when the diet was supplemented with ULB, and approached to that of ABRS, indicating that the effect of ULB on digestibility of RS is similar to that of AB treatment. The digestibility of ABRS was slightly improved by the ULB feeding. Nitrogen retention was highest in lambs fed on ABRS alone, followed by hay with ULB, and was lowest in animals fed on RS with ULB. However, both the amount and proportion of N retention to N intake were enhanced by ULB supplementation to lambs fed on hay. The proportion of N retained to N digested decreased due to ULB supplementation to lambs fed on RS or ABRS. Supplementing ULB did not greatly influence the rumen degradation of either dry matter or crude protein in each of the three diets. RS and hay had similar values in the potential extent of digestion (PED) and digestion rate of PED (kd) of fibrous materials, but the discrete lag time for RS was lower than that for hay. The AB treatment significantly increased the PED (P<0.05) and kd (P<0.05) of RS. Neither the PED nor kd for RS and ABRS was influenced by ULB supplementation, but the kd for hay significantly increased due to ULB. The lag time for hay was also shortened by the ULB feeding. The ULB improved the digestion of fibre in the rumen of lambs fed on low quality roughage. It is inferred that while ULB is effective in increasing nutrient digestibility of low quality roughages by improving ruminal fibre digestion. A synchronized supply of N and energy to rumen microbes should be considered to improve the efficiency of N utilization when the basal diet is ammoniated straw.