Study on Polymorphisms in the Blood Protein of Tibetan Mastiff

[ Objective] The aim was to study the protein polymorphism in the blood of Tibetan Mastiff, and provide some theoretical basis for resource protection and reasonable development and utilization of Tibetan Mastiff varieties. [ Method] A total of 103 blood samples were taken from four populations of Hequ Tibetan Mastiff, Qinhai Tibetan Mastiff, Tibetan Spaniel and native dogs of Qinghai. Seven blood protein Iocus（Tf, Po, Sα2, Hb, AIb, Pr and Amy）were investigated by using vertical polyacrylamide gel electrophoresis with discontinuous buffer system. Then the genetic variation during different populations was analyzed. [ Result] Genetic variations were observed in Tf, Sα2 and Po in four populations, others were not polymorphic. There were three alleles at the locus of Tf and Po, two alleles at the loci of Sα2. Effective number of alleles and Nei＇s average expected heterozygosity were 1. 532 4 and 0.230 3 relatively, all higher in Tibetan Mastiff than other populations. [ Conclusion] Protein locus in blood of Tibetan Mastiff existed in genetic variation.

Optimization of Chelation Process for Complex Microelement Iron Supplement Derived from Pig Blood by Response Surface Methodology

[Objectives]This study aimed to optimize the chelation process for complex microelement iron supplement derived from pig blood by response surface methodology.[Methods]On the basis of single-factor test,p H value,concentration of polypeptide solution and volume ratio of polypeptide solution to FeCl_2 solution were selected as influencing factors with Fe(II)chelation rate as the indicator for Box-Behnken central composite experimental design with three factors and three levels.The effects of three factors on the response value were analyzed by response surface methodology.[Results]The optimized chelation process for complex microelement iron supplement derived from pig blood by response surface methodology was as follows:pH 5.40,polypeptide solution concentration 2.27%,volume ratio of polypeptide solution to FeCl_2 solution 2.16∶1.Under this condition,the predictive Fe(II)chelation rate of iron supplement was 79.37%,while the actual value was 79.41%.[Conclusions]The optimized process may provide new thoughts for the development and utilization of complex microelement iron supplement derived from pig blood.

The Cross-talk between ROS and p38MAPKα in the Ex Vivo Expanded Human Umbilical Cord Blood CD133^+ Cells

This study investigated the correlation between and compared the effects of reactive oxygen species（ROS） and p38 mitogen-activated protein kinase α（p38MAPKα） in the ex vivo expanded umbilical cord blood（hUCB） CD133＋ cells.hUCB CD133＋ cells were cultured in the hematopoietic stem cells（HSCs） culture medium with N-acetylcysteine（NAC,an anti-oxidant）,p38MAPKα-specific inhibitor（SB203580） or their combination.The levels of ROS and expression of phosphorylated p38MAPKα（p-p38） in CD133＋ cells were flow cytometrically detected.The efficacy of ex vivo expansion was evaluated by the density of CD133＋ cell sub-group colony-forming cells（CFC） and cobblestone area-forming cells（CAFC） assay.Our results showed decreased ROS levels in NAC,SB203580,and their combination treatment groups were almost 37%,48%,and 85%,respectively.Furthermore,SB203580 abrogated the activation of p38MAPKα more obviously than NAC.Moreover,the CD133＋ cells in SB203580 treatment group had a 21.93±1.36-fold increase,and 14.50±1.19-fold increase in NAC treatment group,but only 10.13±0.57-fold increase in control group.In addition,SB203580 treatment led a higher level increase in the number of CFU and CAFC than NAC did.These findings suggested that,in expanded CD133＋ cells,ROS activates p38MAPKα,which,in turn,induces ROS production,and p38MAPKα might be the most suitable regulator in ROS-p38MAPKα pathway for the promotion of HSCs ex vivo expansion.

Comparison of the level of thrombus precursor protein in blood plasma between patients with acute cerebral infarction and healthy persons at different time

BACKGROUND： Thrombus precursor protein （TpP） is the index of thrombus activity level, and it is also early referencing index in detecting thrombus diseases. OBJECTIVE： To dynamically observe the changes of TpP level in blood plasma of patients with acute cerebral infarction at different time after onset, and to compare the differences of plasma TpP level between patients with acute cerebral infarction and healthy persons who received health examination. DESIGN： Controlled observation SETTING： Department of Neurology, Affiliated Hospital of Xuzhou Medical College PARTICIPANTS： Totally 58 patients with acute cerebral infarction who received the treatment in the Department of Neurology, Affiliated Hospital of Xuzhou Medical College between September 2004 and March 2005 were recruited in this study. They all met the diagnostic criteria revised by the 4^th National Conference of Cerebrovascular Disorders in 1995 and were diagnosed by clinical and skull CT and （or） MRI examinations. The patients included 33 male and 25 female aged from 36 to 87 years. Time to onset 〈 6 hours, 6 to 11 hours, 12 to 23 hours, 24 to 48 hours and 〉 48 hours were found in 10,11,14,10 and 13 patients respectively. Another 51 persons who homeochronously received the health body examination in our hospital were recruited, including 34 male and 17 female, aged 38 to 85 years, serving as control group. Patients with cardio-cerebrovascualr diseases or liver and kidney diseases were excluded. All the involved subjects were informed of the detected items. METHODS： About 4 mL venous blood was respectively taken from patients admitted to the hospital within 6 hours, 6 toll hours, 12 to 23 hours, 24 to 48 hours and more then 48 hours after onset, and healthy persons when receiving health examination. The level of TpP in blood plasma was measured with enzymelinked immunosorbent assay. MAIN OUTCOME MEASURES： ① Comparison of the level of plasma TpP between patients and controls;② Comparison of the level of plasma TpP of patients with acute cerebral infarction at different time after onset. RESULTS： Totally 58 patients with acute cerebral infarction and 51 persons who received health examination participated in the result analysis. ①Comparison of plasma TpP level between patients and controls： The plasma TpP level of patients with acute cerebral infarction was significantly higher than that of control group [（16.12±3.28）vs （5.38±1.36） mg/L, t= 20.993, P〈 0.01 ]. ② Comparison of plasma TpP level of patients with acute cerebral infarction at different time after onset： The level of plasma TpP was （12.06±3.06） mg/L within 6 hours, （15.11±3.42） mg/L at 6 to 11 hours, （20.63±4.05） mg/L at 12 to 23 hours, （16.15±3.50） mg/L at 24 to 48 hours and （11.88±3.11） mg/L at more than 48 hours after onset. It increased from the 6^th hour, reached the peak at the 12^th to 23^rd hours, maintained at very high level at the 48= hour and then gradually decreased and recovered to the level within 6 hours after onset. The level of plasma TpP of patients with acute cerebral infarction was signiticantly higher at the 12^th to 23^rd hours after onset and the 24^th to 48^th hours after onset than within 6 hours after onset （t = 13.385, P 〈 0.05）. CONCLUSION： ①The level of plasma TpP of patients with acute cerebral infarction is significantly higher than that of persons who received health examination.② Plasma TpP levels of patients with acute cerebral infarction change in wave manner at the different time after onset.

Monitoring the changes in plasm C-reactive protein,fibrinogen and blood white cell in patients with primary hypertension combined with acute cerebral infarction

BACKGROUND： Inflammatory reaction and the increased level of its accompanying active protein play an important role in the occurrence and development of cerebral infarction. C-reactive protein, fibrinogen and white blood cell, as the monitoring index of inflammatory reaction, are very important in the occurrence and development of acute cerebral infarction. OBJECTIVE： To make a comparison between patients with primary hypertension accompanied with acute cerebral infarction and with simple primary hypertension by observing the changes in plasma C-reactive protein and fibrinogen levels as well as white blood cell and differential counts and analyzing their significances. DESIGN ： Controlled observation SETTING ： Ward Building for VIP, Shenzhen Hospital, Peking University. PARTICIPANTS： Totally 133 patients with primary hypertension were selected from Ward Building for VIP, Shenzhen Hospital, Peking University during September 2003 to September 2005, The diagnostic criteria were based on the hypertension diagnosis criteria formulated by the 7^th World Health Organization-International Society of Hypertension Guidelines （WHO-ISH） in 1998. The informed consents were obtained from all the participants. The involved patients were assigned into two groups： primary hypertension group, in which, there were 65 patients with primary hypertension （ degree 2）, including 42 males and 23 females, with mean age of （61 ±14）years and mean blood pressure of （162.7±6.8）/（94.2±8.4） mm Hg （1 mm Hg =0.133 kPa）, and primary hypertension combined with cerebral infarction group, in which, there were 68 patients with primary hypertension combined with cerebral infarction （ meeting the diagnostic criteria formulated in the 4^th National Cerebrovascular Diseases Meeting in 1995 and diagnosed by skull CT or MRI to exclude the patients with lacunar infarction）, including 42 males and 26 females, with mean age of （56±15） years and mean blood pressure of （176.4±9.2）/（96.3±9.7） mm Hg. METHODS： Plasm C-reactive protein and fibrinogen levels, and white blood cell and differential counts of patients in the two groups were examined 24 hours after stroke. The above indexes were re-examined in the primary hypertension combined with cerebral infarction group 72 hours after stroke. White blood cell and differential counts were performed with laser method （East Asia FE-95001 RAM-1, Japan）. The level of C-reactive protein was measured with turbidimetry （BNII Automatic Systems For Analysis, USA）. The level of fibrinogen was measured with algorithm method when prothrombin time was normal and with Clauss method when prothrombin time was abnormal （ACL Automatic Coagulation Analyzer, USA）. MAIN OUTCOME MEASURES： The plasm C-reactive protein and flbrinogen levels, and white blood cell and differential counts 24 hours after stroke in two groups and 72 hours after stroke in primary hypertension combined with cerebral infarction group. RESULTS： All the 133 involved patients participated in the result analysis. The plasm C-reactive protein and fibrinogen levels, and white blood cell and neutrophil counts in patients with primary hypertension were all within the normal range. The plasm C-reactive protein and fibrinogen levels, and white blood cell and neu- trophil counts in patients with primary hypertension combined with cerebral infarction were significantly higher than those in patients with primary hypertension 24 hours after stroke and 72 hours after stroke respectively[24 hours after stroke：（32.12±11.76） mg/L vs. （5.02±3.21 ） mg/L;（4.64±0.75） g/L vs. （3.12±0.49） g/L; （9.32±81）×10^9 L^- 1 vs. （5.78±1.32）×10^9L^- 1 （7.85±2.38）×10^9 L^- 1 vs.（3.49±1.28）×10^9 L^-1,t =7.094, 5.759,4.106,5.491, respectively,all P〈 0.01; 72 hours after stroke： （47.62±18.43） mg/L vs. （32.12±11.76） mg/L; （5.08±0.82） g/L vs. （4.64±0.75） g/L, t =2.864,2.220, respectively, both P 〈 0.05]. CONCLUSION： The increase in fibrinogen level and white blood cell count are the important index in monitoring primary hypertension combined with acute cerebral infarction. The increase in plasm C-reactive protein and fibrinogen levels 72 hours after stroke indicates that plasma C-reactive protein and fibrinogen are very important in the development of disease.

Adenovirus-mediated expression of pig α(1,3) galactosyltransferase reconstructs Gal α(1,3) Gal epitope on the surface of human tumor cells

Gal alpha(1, 3) Gal (gal epitope) is a carbohydrate epitope and synthesized in large amount by alpha(1, 3) galactosyltransferase [alpha(1, 3) GT] enzyme on the cells of lower mammalian animals such as pigs and mice. Human has no gal epitope due to the inactivation of alpha(1, 3) GT gene but produces a large amount of antibodies (anti-Gal) which recognize Gal alpha(1, 3) Gal structures specifically. In this study, a replication-deficient recombinant adenoviral vector Ad5sGT containing pig alpha(1, 3) GT cDNA was constructed and characterized. Adenoviral vector-mediated transfer of pig alpha(1, 3) GT gene into human tumor cells such as malignant melanoma A375, stomach cancer SGC-7901, and lung cancer SPC-A-1 was reported for the first time. Results showed that Gal epitope did not increase the sensitivity of human tumor cells to human complement-mediated lysis, although human complement activation and the binding of human IgG and IgM natural antibodies to human tumor cells were enhanced significantly after Ad5sGT transduction. Appearance of gal epitope on the human tumor cells changed the expression of cell surface carbohydrates reacting with Ulex europaeus I (UEA I) lectins, Vicia villosa agglutinin (VVA), Arachis hypogaea agglutinin (PNA), and Glycine max agglutinin (SBA) to different degrees. In addition, no effect of gal epitope on the growth in vitro of human tumor cells was observed in MTT assay.

Influencing factors of rat small intestinal epithelial cell cultivation and effects of radiation on cell proliferation

INTRODUCTIONCrypt epithelial cells in normal small intestineproliferate at a high speed. But they are verydifficult to culture in vitro and passage stably. A lotof studies have been done[1-16]. Some domestic labsisolated and cultured crypt cells from embryonalintestines and aseptic animal intestine, but failed.We introduced normal rat epithelial cell line-IEC-6from the USA and its living condition for stablepassage was successfully established after trials. Thecell line was testified to be the small intestinalepithelial cell by electron microscopy,immunihistochemistry and enzymatic histoch-emistry. It has been applied to some relatedresearch work[17-21]. It was found that manyfactors were involved in the culture system. Ourpresent study focuses on the culture method and theinfluencing factors on IEC-6.

Relationship between Two Blood Stasis Syndromes and Inflammatory Factors in Patients with Acute Coronary Syndrome

Objective： To investigate the relationship between inflammatory factors and two Chinese medicine（CM） syndrome types of qi stagnation and blood stasis（QSBS） and qi deficiency and blood stasis（QDBS） in patients with acute coronary syndrome（ACS）. Methods： Sixty subjects with ACS, whose pathogenesis changes belongs to qi disturbance blood stasis syndrome, were divided into 2 groups： 30 in the QSBS group and 30 in the QDBS group. The comparative analysis on them was carried out through comparing general information, coronary angiography and inflammatory factors including intracellular adhesion molecule-1（ICAM-1）, chitinase-3-like protein 1（YKL-40） and lipoprotein-associated phospholipase A2（Lp-PLA2）. Results： Compared with the QSBS group, Lp-PLA2 and YKL-40 levels in the QDBS group showed no-significant difference（P〉0.05）; ICAM-1 was significantly higher in the QDBS group than in the QSBS group in the pathological processes of qi disturbance and blood stasis syndrome of ACS（P〈0.05）. Conclusion： Inflammatory factor ICAM-1 may be an objective basis for syndrome typing of QSBS and QDBS, which provides a research direction for standardization research of CM syndrome types.

Effect on Life Span and Membrane Protein in Red Blood Cells by Integrated Medicine Therapy on Chronic Aplastic Anemia

Objective: To explore the mechanism ofintegrated traditional Chinese and Westernmedicine (TCM--WM ) therapy on chronicaplastic anemia (CAA). Methods: The RBClife span of 30 normal human subjects and 30patients with CAA were measured by sir labelled technique before and after TCM--WMtherapy. The morphology and distribution ofRBC membrane protein granules were observed by freeze fracture etching and transmission electron microscope. Results: The halflife of erythrocytes (RBC TI/2)was shortenedin CAA cases and there was a significant difference compared to healthy control (P <0. 01). After therapy, the RBC life span prolonged and approached the normal level. Before treatment, there existed abnormal in morphology, decrease in amount and uneven indistribution of protein granules in protoplasmicface (PF) and extracellular face (EF) of RBCmembrane. After treatment, the protein granules of RBC membrane was improved and approached to control. Conclusions: The morphology, amount, quality and distribution ofRBC membrane protein granule were closelyrelated to its life span. The therapeutic effectof TCM--WM was better than that of WMalone and it had a function both in stabilizingmembrane protein and extending the RBC lifespan.

Rehabilitation therapy for short bowel syndrome

OBJECTIVE: To investigate the effect of rehabilitation therapy for short bowel syndrome on patient nutritional status and intestinal adaptation. METHODS: The rehabilitation therapy included enteral or parenteral nutrition, glutamine, recombinant human growth hormone and rehabilitative diet. From January 1997 to July 2000, twenty - seven patients with short bowel syndrome received the treatment. The average age of the patients was 38.5 +/- 19.3 years, and the length of residual small intestine ranged from 15 to 80 cm, with an average of 46.8 +/- 23.4 cm. The ileocecal valve was preserved in 14 cases, and the average time between the onset of short bowel syndrome and the rehabilitation therapy was 86 +/- 105 days. RESULTS: After the treatment, nutritional status of the patients improved markedly, and intestinal absorptive capacity improved. Eight patients were followed up for more than 2 years, among whom 4 (50%) were weaned from total parenteral nutrition. Thirteen patients were followed up for more than 1 year, and 10 patients (76.9%) were weaned from total parenteral nutrition. CONCLUSIONS: Rehabilitation therapy for short bowel syndrome can improve patient nutritional status effectively and promote intestinal adaptation, providing a new hope for these patients. The therapeutic effects are related to the length of the residual small intestine, patients age and duration between massive intestinal resection and start of the treatment. Early initiation of rehabilitation therapy promotes intestinal adaptation and increases patients ability to wean from total parenteral nutrition.