Rabbit antisera against mouse LDH-C_4 were generated by immunization of rabbits with highly purified mouse lactate dehydrogenase C_4. Immunofluorescent localization of LDH-C_4 on mouse and human sperms was performed by rabbit antibodies against mouse LDH-C_4 at 1:20 dilution of antisera. Sheep anti-rabbit IgGFITC conjugates at dilution of 1:10 was used as second antibody for the detection. Human sperms were obtained from fresh semen of fertile males. Mouse sperms were prepared from epididymis fluid of mature mice. Both human and mouse sperms were thoroughly washed by PBS buffer. Sections of 4 ?m mouse testes were made and dried on the slides for the immunohistochemical localization, ?-hydroxy-n-valeric acid was used as specific substrate in incubation medium for revealing the LDH-C_4 activity. Immunofluorescent studies demonstrated that LDH-C_4 can be found on the surface of mouse and human spermatozoa. In human spermatozoa, the strong fluorescence was seen on the neck and midpiece. On mouse sperm, the antibody binding was directed toward the tail segment with little or no binding on the head or midpiece. Histochemical studies showed that the intensive formazen deposits were on the neck and midpiece of human and mouse spermatozoa, as well as the cells approximate to the lumen of mouse seminiferous tubules.

Objective To study the value of the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in gene diagnosis on spinal muscular atrophy (SMA).Methods PCR-RFLP method was used to detect the homozygous deletion of the exon 7 or exon 8 of SMN gene in 20 SMA patients of Type Ⅰ,Ⅱ,Ⅲ and 15 normal individuals.Results Homozygous deletion of exon 7 and exon 8 of the SMN gene were all identified 7/7 in SMA TypeⅠpatients, and 5/5 and 4/5 respectively in SMA Type Ⅱ patients, but only 1/8 of SMA Type Ⅲ patients, and no homozygous deletion was found in the normal controls.Conclusions PCR-RFLP might be recommended as an effective diagnosis for spinal muscular atrophy Type Ⅰand Ⅱ patients, whereas the method might not be as useful in Type Ⅲ as in Type Ⅰand Ⅱ for the gene diagnosis.

Objective To explore the relationship between angiogenesis and ischemic neurological deficit repair through exogenous interventions,and investigate optimal intervention routes for angiogenesis in cerebral ischemia rats.Methods Totally 240 SD rats were randomly divided into sham operation group,ischemic control group,ischemia and vascular endothelial growth factor(VEGF) group [1 653.3 ng/(g?d),i.p.,for 10 d],ischemia and endothelial progenitor cell transplantation(EPCs) group(2?1010/L,10 ?l),ischemia and anti-VEGF monoclonal antibody group [3 mg/(kg?d),i.p.,for 10 d].Cerebral microvessel density,somatosensory evoked potentials(SEP) and motor evoked potentials(MEP) and behavior were observed in 1,3,7,14,30 and 60 d after injury.Results New blood vessel density of ischemia and VEGF group,ischemia and EPCs group were significantly higher than ischemic control group in all time points besides 1 d after injury(P

Objective To establish a large animal ( sheep) model to serve the experiments of domestic pulsatile ventricular assist device.Methods Three small-tail Han-sheep were anesthetized and the vein access and artery access were achieved.The cardiopulmonary bypass was established through left thoracotomy.Ventricular fibrillation was induced. An hole was made in the apex of left ventricle and the apex cannulation was sutured to it.The aortic cannulation was su-tured to the descending aorta.The two cannulations were connected to the domestic pulsatile ventricular assist device ( DP-VAD) and the driver was turned on.The working of DPVAD and the conditions of the animals were observed.Results The DPVAD worked well and uni-directional blood flow was driven by positive and negative pressure.The left ventricle was unloaded and the blood pressure was raised up.Conclusion The establishment of sheep model of pulsatile ventrieular as-sist device may play important role for the research and development of DPVAD in our country.

Currently,the toxicity study of traditional Chinese medicine is faced with the following problems. Firstly,the evaluation in vitro cannot fully reflect the true state of the body. Secondly,the traditional method is not sensitive enough to the early toxicity. Lastly,the toxicity evaluation indexes cannot determine whether the compatibility of traditional Chinese medicine produces toxicity or increases toxicity systematically. The paper proposed a synthesized early evaluation research method for target organ toxicity induced by traditional Chinese medicine:screening,validation,optimization and application. This method mainly inoolves early target organ toxicity biomarkers in screening,optimi?zation,validation,biological significance explanation,and application to the traditional Chinese medicine incompatibility based on the metabolic dynamic fingerprint spectrum in order to obtain biomarkers of target organ toxicity that are sensitive and precede conventional biochemical indices for early evaluation . We attempted to analyze the pattern of chang of the biomarkers for animals acted by″18 incompatible medicaments″compatibility combination. We found that Radix Aconiti Lateralis Preparata with cardiotoxicity were compatible with Rhizoma Pinelliae,and that Trichosanthes kirilowii Maxim,Fritillaria,Ampelopsis Radix and Bletilla striata without non-cardiotoxicity produced and increased cardiotoxicity systematically.

Objective To study the effects of improving elderly patients with bowel dysfunction after cardiopulmonary bypass.Methods Choose 54 elderly patients with CPB were randomly divided into tree groups:the control group and the Gln group and the Combination group,every group has 18 patients.Glutamine(Gln) at 0.4 g/kg was administrated to the patients in the Gln group and the combination group during the beginning at CPB and two days after the operation.Based on the Gln group,the patients of The Combination group were given the different oral Chinese medicine before and after operation,according to syndrome differentiation classification.Plasma endotoxin,IL-6 and DAO were measured before operation and at the endof operation and 24 h,48 h after operation.At the same time,we used German Siemens Company's X3000 Color Doppler machine to test every group of patients superior mesenteric artery.We tested before surgery,and one to two days after operation,from the starting point within 2cmof the superior mesenteric artery and the angle between the ultrasound beam flow was less than 60 °,the probe frequency was 3.5 MHz.The detection of hemodynamic indexes were EDV and PI.Results After the test,the plasma endotoxin and IL-6 levels and DAO levels of the Gln group were reduced than the Control group after cardiopulmonary bypass.The plasma endotoxin and IL-6 levels and DAO levels of the Combined gnup were decreased than the Gln group after cardiopulmonary bypass and one day after operation.Analyze superior mesenteric artery diastolic velocity (EDV)and pulsatility index (PI) by the Color Doppler.After operation,the EDV in the Gln group and the Combined grup were significantly higher than the control group,the Pl were lower than the control group.The parameters levels have significant difference after cardiopulmonary bypass and one day after operation(P ＜ 0.05 ).The Combined group was improved than the Gln group after cardiopulmonary bypass and one day after operation.According to the traditional Chinese medicine diagnosis,there were eighteen patients in the Combined group,including fifteen patients in type of deficiency of both qi and yin and three patients in type of fu-viscera syndnome.All of the patients have defecated within tree days after operation.In the type of deficiency of both qi and yin nine patients were cured and six were shown to be effective after operation.All the patients in type of fuviscera syndrome were cured.Conclusion N(2)-L-Alanyle-L-Glutmaine have protective effect on the intestinal mucosal barrier after CPB.Combined traditional Chinese Medicine preparation was more conducive to the intestinal function recovery of elderly patients after cartiopulmonary bypass.

Objective To investigate the clinic effect of the Salvia miltiorrhiza combined with dextran to prevent veno-occlusive disease after hematopoietic stem cell transplantation. Methods In the process of the pretreatment of the hematopoietic stem cell transplantation, patients were treated with salvia miltiorrhiza (20 ml/d), dextran(250 ml, twice a day) by venous transfusion and the drugs to protect the liver cell was used in the same time. When the count of platelet dropped to 30×109/L, salvia miltiorrhiza and dextranware stopped applying forever. Results Veno-occlusive disease and hemorrhage has not occurred during 85 times of the hematopoietic stem cell transplantation treated with salvia miltiorrhiza and dextran. Conclusion We conclude that the combined treatment with salvia miltiorrhiza and dextran is safe and effective to prevent veno-occlusive disease after hematopoietic stem cell transplantation.

Objective To establish an animal model for implantable ventricular assist device. Methods Seven male calves, body weight 120 to 180 kg, were anesthetized and underwent surgical implantation through a left thoracotomy at the 4th intercostal space. The domestic-made implantable ventricular assist device (DIVAD) was implanted after cardiopulmonary bypass and induction of ventricular fibrillation. The major indexes of mechanical properties of the DIVAD are similar to those of imported devices. The DIVAD was 29.5 mm × 72 mm, with a total weight of 158 g. The maximal pump speed was 9000 rpm and the maximal pump flow was 8 L/min in in vitro experiment. A flow meter was incorporated into the pump. The DIVAD was constructed with an external controller, and the pump can be monitored and adjusted through it. The inflow cannula was positioned into the left ventricle and the outflow cannula was sutured to the descending aorta. The pump was run at 3,500 to 8,000 rpm. The cardiopulmonary bypass was weaned. The pump output and animal condition was continuously monitored. Heparin was administered to maintain the activated clotting time at 1.5 to 2 times its baseline value. Results All the seven calves weaned from cardiopulmonary bypass. The mean survival time is 20.28 hours with a range from 0.5 to more than 93 hours. Conclusion Calf is the right animal model for NIVAD implantation and there are some characteristics in its perioperative treatment. The establishment of this animal model may play important role for further improvement of NIVAD.

AIM: G protein-coupled inwardly rectifier potassium (GIRK) channel are distributed widely in mammalian brain. In CNS, GIRK 1/2 seems to be the predominant heterotetramers which play a pivot role in the regulation of the excitability of neurons and may contribute to the resting potential by leading to a hyperpolarization of membrane potential and reduction of the action potential frequency. In the context, the Weaver mouse is the first neurological abnormality directly linked to a genetic point mutation in the GIRK2 protein which includes spontaneous seizure. GIRK2 knock out mice showed normal development but more susceptible than normal mice to seizure induced by GABA antagonist. Here, we report that the mRNA and protein expression of GIRK subunit 2 is altered in kainic acid(KA)-induced epileptic rat hippocampus. METHODS: Rats were injected with kainate 14 mg/kg intraperitoneally to establish an acute and chronic temporal lobe epilepsy model. At chronic spontaneous seizure stage, by using of in situ hybridization, immunocytochemistry and Western blotting, the GIRK 1,2 mRNA and protein were analyzed quantitatively in the dentate gyreus, CA1, CA3 regions of hippocampus. RESULTS: GIRK1,2 mRNA and proteins were expressed abundantly in all regions of hippocampus. KA induced seizures and caused a significant increase in GIRK2 mRNA abundance and immunoreacitivity; only GIRK1 mRNA was increased significantly, but no difference was found by Western blotting protocol. CONCLUSION: GIRK1,2 mRNA and protein expression in the hippocampus of epileptic rat brain is up-regulated, which may be an adaptive response to over-excitability of neuron networks and prevent the over-excitability spread in hippocampus (DG-CA3-CA1). [

Objective: To observe the effects of ginger-partitioned moxibustion at Shenque (CV8) and Guanyuan (CV4) on the expression levels of endocrine-related molecules and their receptors in rats with primary dysmenorrhea (PD) due to cold-dampness stagnation, thus to explore their analgesic mechanisms. Methods: Thirty-two female Wistar rats were divided into a normal group, a model group, a ginger-partitioned moxibustion group, and a Western medicine group according to the random number table method, with 8 rats in each group. Except for rats in the normal group, all other rats were treated with oxytocin combined with ice-water bath to establish the rat models of PD due to cold-dampness stagnation. After successful modeling, rats in the normal group and the model group did not receive treatment; rats in the ginger-partitioned moxibustion group received treatments with ginger-partitioned moxibustion at Shenque (CV8) and Guanyuan (CV4); rats in the Western medicine group received ibuprofen by intragastric administration. The writhing response of rats was compared among groups, and the serum levels of prostaglandin F2α (PGF2α), estrogen (estradiol, E2), progesterone (P), and the mRNA expression of PGF2α and E2 receptors in the uterine tissues were detected. Results: No writhing behavior was observed in the normal group; compared with the normal group, the serum PGF2α and E2 levels in the model group were increased (P<0.01), while the P level was decreased (P<0.01), and the mRNA expression levels of the uterine PGF2α and E2 receptors were increased (P<0.01, P<0.05). Compared with the model group, the writhing behavior latency was prolonged, and the writhing response score was decreased in the ginger-partitioned moxibustion group and the Western medicine group (P<0.01); the serum PGF2α and E2 levels in the ginger-partitioned moxibustion group and the Western medicine group were decreased, while the P level was increased (P<0.05 or P<0.01); the mRNA expression levels of the uterine PGF2α and E2 receptors in the ginger-partitioned moxibustion group and the Western medicine group were decreased (P<0.05). Compared with the Western medicine group, the ginger-partitioned moxibustion group showed a prolonged writhing behavior latency, reduced writhing response score (P<0.05), and decreased serum E2 level (P<0.05), while no statistical differences in the serum PGF2α and P levels, or the mRNA expression levels of uterine PGF2α and E2 receptors (P>0.05).Conclusion: The analgesic effect of ginger-partitioned moxibustion on PD due to cold-dampness stagnation may be related to regulating the mRNA expression levels of PGF2α and E2 receptors in the uterine tissues.