OBJECTIVE: To evaluate the therapeutic effect of transplantation of autologous bone marrow mononuclear cells combined with traditional Chinese medicine for the treatment of limb ischemia. METHODS: Twenty-three patients with limb ischemia were treated. G-CSF was used to stimulate the bone marrow. The mononuclear cells were separated from the aspirated bone marrow fluid in the stem cell studio. The cell amount was above 1x10(9). The transplantation was performed by the way of intra-muscular multi-injection. Traditional Chinese medicine for replenishing qi to activate blood was prescribed from the first day after operation. The pain, poikilothermia, ulcer or necrosis and ankle/brachial index (ABI) of the ischemic limb were evaluated before and after the treatment. RESULTS: The pain score and poikilothermia score decreased one month after the transplantation, with distinct differences as compared with the scores before the treatment (P<0.05). The ABI increased gradually after the treatment, and one month after the treatment, it was 0.15 higher than that before the treatment. CONCLUSION: Transplantation of autologous bone marrow mononuclear cells combined with traditional Chinese medicine can decrease the symptoms and signs of severe lower limb ischemia effectively, and improve the circulation of the ischemic area.

To observe the effects of tetramethylpyrazine (TMP) on the proliferation and type I collagen synthesis of rat cardiac fibroblasts (CFBs) induced by angiotensin II (Ang II), and to explore the mechanism of TMP in treating myocardial fibrosis.

Objective:To test the expression of Minichromosome maintenance complex component 7(MCM7) protein in hepato-cellular carcinoma(HCC) of different species including human, rat and tree shrew (tupaia) by cross-species oncogenomics approach, and to investigate the relationship between the expression of MCM7 and the development of hepatocellular carcinoma and its clinical significance. Methods:Western blot and Immunohistochemistry were applied to detect the expression levels of MCM7 protein in HCC tissues,corresponding HCC-adjacent liver tissues and normal liver tissues collected from different species including human, rat and tree shrew, respectively. The clinicopathologic factors were also analyzed with the results of Immunohistochemistry. Results:Western blot analysis showed that the expression of MCM7 protein in HCC tissues of human and rat were higher than that in corresponding HCC-ad-jacent liver tissues and normal liver tissues, respectively and significantly (P<0.05). However, the expression of MCM7 protein in HCC tissues of tree shrew were also higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, but no significant difference was found among three types of tissues (P>0.05).There was also no significant difference between HCC-adjacent liver tis-sues and normal liver tissues in three species (P>0.05). Immunohistochemical analysis showed that MCM7 protein was mainly ex-pressed in nucleus of HCC cells, and the positive rate of MCM7 protein in HCC tissues of human, rat and tree shrew were significantly higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, respectively (P<0.05). However, no significant difference was found between HCC-adjacent liver tissues and normal liver tissues (P>0.05). Moreover, the protein level of MCM7 was intimately related to patient's HCC stage, extrahepatic metastases and postoperative recurrence (P<0.05). Conclusion:MCM7 protein might play a pivotal role in hepatocarcinogenesis. In addition, it was probably related to patient's HCC stage, extrahepatic metastases and postoperative recurrence. It seems very likely that MCM7 may be applied as a new molecular target in HCC prevention and treat-ment.

Objective To investigate the method to isolate and culture hepatic stellate cells ( HSCs) for studying the cellular mechanisms of hepatic frbrosis.Methods HSCs were isolated by nycodenz density gradient centrifugation after the hepatocytes obtained from adult male gebils were digested with pronase, collagenase and DNase, infused via portal vein.The cell viability was determined by trypan blue exclusion test.The purity of HSCs was identified by detectingα-SMA, desmin immunohistochemical staining.Results The yield rate of HSCs was 0.5~1 ×107 per gerbil liver, and the cell viability was more than 90%.The percentage ofα-SMA-positive cells was more than 75%after 3 days primary culture and almost 100% cells were α-SMA and desmin positive in passage culture.Conclusion The successful protocol of primary culture of Mongolian gerbil HSC provide a technical support for research of relevant liver diseases and drug development in the future.

Objective To investigate the numbers of corpus luteum and ovarian follicles and compare the levels of serum prolactin (PRL), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and estradiol (E2 ) in different phases of estrus cycle in female gerbils .Methods Consecutively taking vaginal smears of the gerbils and directly examined under light microscope to distinguish the four phases of the estrus cycle .Hematoxylin and eosin staining was used to histological examination of the gerbil ovaries , and to detect the levels of serum PRL , LH, FSH and E2 by ELISA assay during estrus cycle .Results The proportion of cornified vaginal exfolliated cells could be the basis to distinguish four phases respectively:proestrus, oestrus, metoestrus, and dioestrus.Moreover, there were no significant differences between the numbers of ovarian follicles in different phases of estrus cycle .The numbers of corpus luteum in preoestrus were significantly lower than that in the other phases of estrus cycle ( P <0.05 ) .The levels of serum PRL and LH were increasing constantly from preoestrus to dioestrus , and both reached a peak at dioestrus ( P<0.05 ) .The levels of serum FSH and E2 both peaked at preoestrus , and were significantly higher than those at oestrus , metoestrus and dioestrus ( P<0.05).Conclusions There are no significant differences between the numbers of ovarian follicles in different phases of estrus cycle .Gonadotropin , prolactin and estradiol paly important roles in the regulation of estrous cycle .The phases during which surges of FSH and E 2 occur in Mongolian gerbils are similar to those of rats and mice , while the PRL and LH are different .Our findings provide further reference to the study of reproductive physiology of Mongolian gerbils .

Objective To explore the basic ingredients of the tree shrew’ s( Tupaia belangeri) milk and compare with the dairy ingredients of other milks.Methods We select ten seed tree shrews after delivery ( 1 ~21 ) d with lactation mother tree shrews, and use artificial passive breastfeeding method let the young tree shrews suck breast milk,we took the milk from the young tree shrews in the stomach, directly using aseptic operation with a syringe immediately, once every two days, for consecutive three to five times, and a total of 18 mL milk was taken from each seed tree shrew.Then the milk was detected according to the national standard method for component testing.Results The total solid content of the tree shrew’ s milk was 43.63%, including 26.01%of fat, 10.41%of protein, 0.45% of lactose and 0.99%of ash content.Compared with cow's milk, the tree shrew’ s milk contained 3.36 times of total solid contents, 1.24 times of ash, 2.74 times of protein, 6.67 times of fat, and 0.09 times of lactose.Compare with baby formula milk, the tree shrew’ s milk contained 1.44 times of total solid contents, 0.20 times of ash, 0.58 times of protein, 1.53 times of fat, and 0.06 times of lactose.The trace mineral composition of the tree shrew’ s milk showed that the calcium, phosphorus, potassium, sodium, magnesium, and iron contents were 1.83 times, 2.73 times, 1.25 times, 1.93 times, 1.28 times, and 1.48 times higher than those in the cow's milk, and were 0.66 times, 0.85 times, 0.34 times, 0.26 times, 0.85 times, 0.24 times lower than those in baby formula milk.Conclusions The main nutrients of tree shrew’ s milk is of high fat, high protein and low sugar, and it can provide a basis for tree shrews artificial brood and breeding work.

Amniotic fluid embolism(AFE) is a fatal intrapartum complication. Typical AFE is characterized by respiratory and circulatory failure, followed by disseminated intravascular coagulation(DIC), while atypical AFE, which had a higher survival possibility, presents with unexplained DIC only. Recent findings support an anaphylactoid inflammatory mechanism underlying the pathophysiology of AFE. Milrinone is currently the first choice for AFE due to its several functions, such as reducing pulmonary artery pressure, and improving right ventricular function through lowering heart rate and positive inotropic action. A multidisciplinary team is required in the rescue of AFE and medical staff who are skilled in critical medicine (such as anesthesiologists) often play a pivotal role.

Objective Cysteinyl leukotrienes are potent inflammatory mediators. Their actions are mediated by specific receptors,the CysLT receptors(CysLT1R and CysLT2R),which have been cloned and characterized. In this stud-y,we investigated the protective effects of the CysLTR antagonist Pranlukast and HAMI 3379 on global cerebral ischemia/reperfusion(CI/R)injury in gerbils and its underlying mechanisms. Methods The gerbil model of CI/R was established by bilateral common carotid artery occlusion for 10 min followed by 24 h reperfusion. Then the animals were equally ran-domized into four groups: sham, model, Pranlukast(0.1 mg/kg)and HAMI 3379(0.1 mg/kg)groups. The later two groups were treated with intraperitoneal injection of Pranlukast and HAMI 3379,respectively,once daily for 4 days before carotid artery occlusion,while the former two groups with saline only,all at 10 mL/kg. After 24 h reperfusion,neurologi-cal deficit scores were observed and the behavioral dysfunction was assessed. The neuron morphology of cerebral cortex and CA1 subregion of hippocampus were observed in brain sections stained with cresyl violet. The expression of autophagy-relat-ed proteins beclin-1 and LC3 in the homogenate of cerebral cortex and hippocampus were determined using western blotting analysis. The ultrastructure of autophagosomes in the CA1 subregion of hippocampus was observed by electron microscopy. Results Compared with the model group, Pranlukast and HAMI 3379 attenuated neurological deficits, improved the be-havioral dysfunction,inhibited the neuron injury and loss, decreased the expression of autophagy-related protein beclin-1 and LC3 and the number of autophagosomes. Conclusions cysteinyl Leukotriene receptor antagonist Pranlukast and HAMI 3379 can alleviate global cerebral ischemia/reperfusion injury in gerbils. The protective effects of Pranlukast and HAMI 3379 appear to be associated with the inhibition of autophagy.

ObjectiveTo explore the histological characteristics of the kidney in Mongolian gerbils of different ages. MethodsTen Mongolian gerbils of 2, 6, and 12 months old (half male and half female) were selected. After euthanasia, the kidneys were taken and paraffin sections were made. After HE, MASSON and PAS staining, the structural differences of different parts of the kidney tissue in gerbils of different ages were observed by digital scanning, and the relevant data of the kidney tissue were measured by image analysis software. ResultsThe number of proximal convoluted tubules was more than that of distal convoluted tubules in the renal cortex and outer medulla of gerbils. With age, the glomerular density decreased, the glomerular diameter increased, the basement membrane of renal tubules thickened, and the fibrous components between renal tubules increased. ConclusionThe histological structure of Mongolian gerbil's kidney varies with age, which may be related to glomerulosclerosis and parenchymal cell reduction. The specific mechanism needs further study.