Chronic myeloid leukemia(CML)is a pluripotent stem cell disease characterized by the proliferation of myeloid cell.Ph chromosome has been found in 95% of CML patients.BCR-ABL fusion protein expressing on Ph chromosome displayed constitutive tyrosine kinase activity.The fusion protein could phosphorylate and activate downstream substrates continually in the process of cell signal transduction,and then cells proliferate and migrate constantly,resulting the occurrence of CML.In recent years, people have been tried various methods to cure CML, which included developing some chemotherapy drugs and herbal extracts,as well as designing and synthesizing the inhibitors targeting BCR-ABL tyrosine kinase.Clinical studies showed that these drugs have been achieved remarkable effects.This paple aimed to review for drugs treated in CML.

OBJECTIVE:To determinate vitamin B1 and vitamin B6 in Gengnianling capsules by HPLC simultaneously .METHODS: The separation was performed on Hypersil-ODS C18 column, methanol - sodium hexanesulfonate solution(20 : 80) was used as mobile phase with a flow rate of 0.8ml/ min and detection wavelength of 280nm.RESULTS: Linear correlations with peak area scores were achieved when the sample size of vitamin B1 and vitamin B6 were with a range of 0.884?g-2.652?g (r = 0.9 999) and 0.714?g-2.142?g(r = 0.9 999) .respectively, the average recovery of which were 95.87%(RSD = 0.82%) and 101.96% (RSD = 0.86%), respectively .CONCLUSION: The method is simple, accurate and it can be used for quality control of Gengnianling Capsule.

ObjectiveTo observe the effect of Xuebijing injection pretreatment on systemic inflammatory response induced by severe heat-stroke, and to investigate the mechanism of alleviation of intestinal injury in rats. Methods Thirty-six healthy adult male Wistar rats with grade SPF were randomly assigned into three groups with randomized number method, namely sham group, severe heat-stroke model group, and Xuebijing pretreatment group (XBJ group), with 12 rats in each group. The animals were placed in a pre-warm chamber [temperature (40±2)℃, humidity (65±5)%] in order to induce typical heat-stroke. The duration of heat-stress was 60 minutes, while the animals in sham group were exposed to ambient temperature of 25℃. Arterial blood samples were collected at the beginning and the end of heat-stress, the concentrations of tumor necrosis factor-α(TNF-α), interleukins (IL-1β, IL-6), and lipopolysaccharide (LPS) in peripheral blood were determined by enzyme linked immunosorbent assay (ELISA). The intestinal tissues were harvested after heat-stress, and the pathological changes in intestine tissues were observed after hematoxylin-eosin (HE) staining and under optical microscope. The pathological injury scores were calculated. Immunohistochemistry was performed to determine inducible nitric oxide synthase (iNOS) expression in intestinal tissue. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Western Blot was used to measure the tight junction protein occludin expression.Results The concentrations of TNF-α, IL-1β, IL-6 and LPS in blood of the rats after heat-stress in model group were significantly higher than those of sham group [TNF-α (μg/L): 443.00±110.10 vs. 98.36±44.61, IL-1β (μg/L): 436.37±163.64 vs. 64.24±16.15, IL-6 (μg/L): 342.70±92.42 vs. 54.40±13.22, LPS (μg/L): 0.68±0.22 vs. 0.09±0.02, allP< 0.01], but the levels of these parameters in XBJ group were significantly lower than those of model group [TNF-α (μg/L):340.45±68.57 vs. 443.00±110.10, IL-1β (μg/L): 191.33±82.78 vs. 436.37±163.64, IL-6 (μg/L): 192.21±37.89 vs. 342.70±92.42, LPS (μg/L): 0.43±0.17 vs. 0.68±0.22, allP< 0.01]. Infiltration of inflammatory cells, necrosis and hemorrhage in intestinal mucosa were found in the intestine of heat-stroke animals in model group. The pathological lesions in XBJ group were milder than those of model group, with a decreased pathological injury score compared with model group (2.10±1.15 vs. 3.20±0.67,P< 0.01). The expression of iNOS and apoptosis of cells in intestinal tissue in model group were increased compared with that of sham group, but they were significantly less marked in XBJ group compared with model group [iNOS (adjustedA value): 0.32±0.15 vs. 0.74±0.17, apoptotic index: 0.23±0.08 vs. 0.56±0.07, bothP< 0.01]. The order of expression for occludin protein from high to low was sham group, XBJ group and model group (A value was 0.96±0.25, 0.62±0.20, 0.33±0.11, respectively). Furthermore, there was significant difference in the expression of occludin protein between model group and both XBJ group and sham group (bothP<0.01).Conclusions Xuebijing injection alleviates inflammation and endotoxemia produced by severe heat-stroke in rats. The mechanism may be related to amelioration of oxidative injury, apoptosis, and dysfunction of tight junction protein occludin expression.

Objective To establish a simple , stableand effective method for the isolation and purification of ABL tyrosine kinase and its mutant ABLT315I.Methods pET-28a vector was inserted in abl gene or its site directed mutagenesis.Then Escherichia coli BL21 competent cells were co-transformed with pGEX6P-1-ptp-1b and pET28a-abl/pET28a-ablc944t .The transformed BL21 cells were incubated, and then were stimulated with Isopropyl-β-D-thiogala-ctopyranoside ( IPTG ) to express ABL tyrosine kinase and its mutant .The ABL tyrosine kinase and its mutant was purified by affinity chromatography and gel filtration chromatography .SDS-PAGE was used to detect the purity and relative molecular weight of ABL tyrosine kinase and its mutant.BCA method was used to determine the concentration of ABL tyrosine kinase and its mutant .Finally, kinase activity of target protein was examined by ATP /NADH coupling method .ResuIts SDS-PAGE showed the high purity of ABL tyrosine kinase and its mutant.The concentration of ABL and ABLT 315I protein was reached 28mg/L of LB and 20mg/L of LB, respectively.Both of the target protein was measured to have good tyrosine kinase activity in vitro .ConcIusion A simple, stable and effective method for the isolation and purification of ABL tyrosine kinase and its mutant was found successfully in the study , which laying good foundation for High Throughput Drug Screening and structure analysis of protein subsequently .

This study was aimed to explore the method to improve the purity of primary myocardial cells from neonatal rats. The myocardial tissues of rats which were born 1-3 days were repeated digested by 0.08% myocardial cells digestive juices. And then, cells were neutralized with DMEM medium which containing 10% fetal bovine serum. The cells were separated with differential sidewall ion. The red blood cell cracking liquid was added to remove the red blood cells. Bromine deoxidization uracil (Brdu) was added to purify the myocardial cells. Then, cells were incubated in the CO2 incubator for two days. The serum-free synchronization was performed for one day. The results showed that the output of myocardial cells from one rat was about 1.2 × 106. The dynamic myocardial cells occupied more than 90%. The purity of myocardial cells was more than 90%. After 3 to 4 days, the cell fusion of myocardial cells was formed with spontaneous rhythm beats. It was concluded that the method can ensure the yield and the activity of the myocardial cells. At the same time, the purity of myocardial cells can also be improved greatly.

Objective To investigate the effect ofulinastatin on severe heat-stroke with acute lung injury induced by severe heat stroke.Methods Thirty severe heat stroke patients were divided into conventional group (n=15) and ulinastatin group (n=15) randomly,with another 80 healthy adults serving as controls.The baseline data such as age,gender,onset period and APACHE Ⅱ scores were recorded and compared between the two groups on admission.Peripheral leucocyte counts,oxygenation index and Murray scores were determined on the 1st,3rd and 5th day.The concentration of inflammatory mediators in bronchoalveolar lavage fluid (BALF) and alveolar macrophage supernatant were detected by enzyme-linked immunosorbent assay (ELISA).Western blotting and real-time PCR were used to measure expression of triggering receptor-1 on myeloid cells (TREM-1) on alveolar macrophages.Furthermore,comparison was made in terms of the ventilation period,ICU stay time and mortality in 28 days between the two groups.Results No differences were found in age,gender,onset period and APACHE Ⅱ scores between the two groups (P＞0.05).Compared with the conventional group,peripheral leucocyte counts and Murray scores in the ulinastatin group significantly decreased on the 3rd and 5th day (P＜0.05,P＜0.01).But oxygenation index was higher in the ulinastatin group than in the conventional group (P＜0.05).The concentration of TNF-α and IL-6 in BALF was lower in the ulinastatin group than in the conventional group (on the 3rd day:P＜0.05,P＜0.01;on the 5th day:P＜0.01,P＜0.01).The concentration of TNF-α and IL-6 in alveolar macrophage supernatant was lower in the ulinastatin group than in the conventional group (on the 3rd day:-P＜0.05,P＜0.01;on the 5th day:P＜0.01,P＜0.05).The expression of TREM-1 protein on alveolar macrophages were lower in the ulinastatin group than in the conventional group (on the 3rd day P＜0.01;on the 5rd day P＜0.05).TREM-1 mRNA was lower in the ulinastatin group than in the conventional group (on the 3rd day:P＜0.05;on the 5th day:P＜0.05).Eventually,the treatment of ulinastatin shorten ventilation period and ICU stay time (P＜0.01,P＜0.05).Nonetheless,it failed to show any improvement in terms of the mortality during 28 days (P＞0.05).Conclusion Our study exhibited that ulinastatin had good effect on the heat stroke patients with acute lung injury and it helped reduce the inflammatory reaction of pulmonary tissues.The underlying mechanism of these effects might lie in its ability to reduce heat stroke-induced inflammatory secretion and expression of TREM-1 on alveolar macrophage.

Human tissue typing methods were employed in developing a porcine allotransplantation model. 23 Chinese Sichuan White Pigs(2-3 months old, 17.5+/-4.6kg, with clear family background) were selected for tissue typing, ABO blood type cross reaction, complement-dependent cytotoxicity (CDC) cross reaction and one way mixed lymphocyte reaction (MLR). 6 pairs of swine that showed better matching results were selected as donors and recipients. Single-kidney orthotopic transplantation was conducted after removing both kidneys of the recipient. Five recipients showed low matching results (MLR ranging from 2175 to 3560, CDC from 1 to 4); of them, 2 died of operation, 2 died of acute renal tubular necrosis and accelerating rejection 4 days after operation respectively, and 1 died of acute renal tubular necrosis 4 days after operation. 6 recipients showed high matching results (MLR ranging from 982 to 1916, CDC from 2 to 4); of them, 1 died of anaesthesia during operation, 3 died of accelerating rejection and acute rejection 2 weeks after operation respectively, 1 had good kidney function, and 1 presented weak rejection 1 week after operation but the kidney function came back to normal afterwards. Human tissue typing methods could be adopted in developing the porcine model. Hyperacute rejection could be avoided by screening ABO blood type, CDC and MLR tests. However, based on these primary data, it was hard to evaluate the predictive values of CDC and one-way MLR for accelerating rejection, acute rejection and graft chronic dysfunction. Further research by expanding experiments in these aspects is still going on.
ABO Blood-Group System ; immunology ; Animals ; Cytotoxicity Tests, Immunologic ; Graft Rejection ; prevention & control ; Histocompatibility Testing ; Kidney Transplantation ; immunology ; Swine ; T-Lymphocytes ; immunology ; Transplantation, Homologous ; immunology

As a famous-region Dao-di Herbs, Ligusticum chuanxiong which mainly grows in the west of the upper reaches of Jinma River in Dujiangyan for a long time. In recent years, the history, species and origin of L. chuanxiong were researched by many scholars. However, the forming pattern of Dao-di herbs of L. chuanxiong has not been reported systematically. Basing on the general principles of the formation of Dao-di herbs, it can be concluded that the forming pattern of L. chuanxiong is the type of two determinants, which are combined both unique ecological environment of genuine regions and advanced cultivation techniques.
China ; Ecology ; Ligusticum ; growth & development

Objective This study was designed to determine the effect of transient receptor potential vanilloid type 4(TRPV4)on angiotensin Ⅱ(Ang Ⅱ)-induced renal injury in TRPV4-null mutant(TRPV4 -/-)mice. Methods The mice were divided into sham group and Ang Ⅱ-treated group. Ang Ⅱ was infused systemically into wild type(WT)and TRPV4 -/- mice via a miniosmotic pump for 4 weeks, and the sham mice were given with normal saline. Systolic blood pressure,urinary excretion of albumin and 8-isoprostane, serum creatinine, and the pathological changes in the kidney tissues were assayed after the 4-week treatment. Results Compared with corresponding sham mice,Ang Ⅱ infusion led to enhanced systolic blood pressure,increased urinary excretion of albumin and 8-isoprostane,increased serum creatinine(P< 0.05),and enhanced glomerulosclerosis degree and renal tubulointerstitial injury index(P< 0.05)in the WT and TRPV4 -/- mice. The result were associated with enhanced collagen levels in the kidney(P< 0.05). All of them were attenuated by the deletion of TRPV4 in the absence of alteration in blood pressure(P< 0.05). Conclusions Deletion of TRPV4 could alleviate renal injury during Ang Ⅱ-induced hypertension, suggesting that TRPV4 may contribute to the pathophysiology of angiotensin Ⅱ-induced renal injury.

Myocardial fibrosis is featured by excessive proliferation of cardiac fibroblast and collagen deposition. There is close relationship between myocardial fibrosis and various cardiovascular disease, and it is potential risk factor for sudden death. At present the precise mechanism remains unclear.Myocardial fibrosis has been found to relate with renin-angiotensin-aldosterone system,cell factor,oxidative stress,inflamatory factor,endothelial function obstacles,Intracellular calcium ion. These factors influences the occurrence of myocardial fibrosis by the same or different pathway.