OBJECTIVEThe aim of this study is to establish a transgenic mouse model for cleft palate relevant gene thyroid transcription factor-2 (TTF-2), which can be used to study palatal shelf development when the expression pattern and regular activation of TTF-2 is altered.

METHODSThe C57BL/6J mouse TTF-2 gene was cloned through polymerase chain reaction (PCR) from the mouse genomic DNA. The TTF-2 gene was inserted into the expression vector pBROAD3-mcs to construct the recombinant expression vector pBROAD3-TTF-2. This expression vector was then microinjected into the male pronuclei of the fertilized mouse ovum. Thus, the TTF-2 transgenic mice model was established. The genotype of the transgenic mice was identified by PCR and Southern blot analysis. Immunohistochemistry identified the consistent expression of TTF-2 gene during its palatal shelf development.

RESULTSTTF-2 genes were microinjected into 982 fertilized ova. A total of 580 two-cell-stage embryos cultured and transplanted into the oviducts of 48 pseudopregnant female mice. Overall, 68 embryos were obtained for analysis. The genotype of the mice was determined through PCR and Southern blot analysis using genomic DNA extracted from tail biopsies of the transgenic fetus. A total of 13 TTF-2 transgenic mice were detected. The expression of TTF-2 gene during the palatal shelf development of the transgenic mice was consistently detected by immunohistochemistry.

CONCLUSIONThe recombinant expression vector pBROAD3-TTF-2 was integrated into mouse genome through microinjection. The transgenic mouse in the palatal shelf that consistently expressed TTF-2 was successfully established and displayed a cleft palate phenotype.

Animals ; Cleft Palate ; Disease Models, Animal ; Female ; Forkhead Transcription Factors ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Thyroid Gland

Objectives To observe the pre and post-operational changes of the expressions of survivin, caspase-3 and CD44V6 in patients with colorectal cancer after intra-abdominal implantation of sustained releasing fluorouracil. Meth-ods Sixty-four patients with colorectal cancer (Dukes’stage of B and C) were divided into treatment group and control group, 32 patients in each group. The standard radical surgery was performed in two groups of patients. The fluorouracil im-plants were implanted intra-abdominally in treatment group. The peripheral blood levels of surviving and caspase-3 were de-tected by RT-PCR. The level of CD44V6 was detected by flow cytometry in two groups of patients. Results There were no significant differences in levels of survivin, caspase-3 and CD44V6 before surgery between two groups (P>0.05). The level of survivin (0.362 ± 0.183) was significantly lower at 14 days after operation in treatment group than that of control group (0.585±0.207), but the level of caspase-3 (2.001±0.146) was significantly higher than that of control group (1.654±0.111). The levels of CD44V6 were significantly lower in treatment group (1.857±0.535) and control group (3.471±0.496) after opera-tion than those before operation (9.557±1.170 and 9.729±0.943, P＜0.05), and the level of CD44V6 was significantly lower in treatment group than that of control group (P＜0.05). Conclusion The implant for the sustained release of fluorouracil showed a positive impact on micrometastases and prognosis of colorectal cancer, while improved the long-term efficacy of postoperative colorectal cancer.

To investigate whether glutamate and voltage-gated calcium channels-independent calcium influx exists during acute anoxic neuronal damage and its possible relationship to neuronal protective function of NGF. In in vitro model of acute anoxia, hippocampal cultures from newborn rats were exposed to 3 mmol/L KCN. Changes of intracellular Ca(2+) concentration ([Ca(2+)](i)) were monitored by con-focal imaging and cell viability was assayed by PI and cFDA staining. The results showed that after treatment with primary hippocampal cultures with 3 mmol/L KCN for 15 min, [Ca(2+)](i) was significantly increased 6.27-fold compared to pre-anoxia level and 73.3% of the cells died. When combination of 20 mumol/L MK-801 (glutamate receptor antagonist), 40 mumol/L CNQX (AMPA receptor antagonist) and 5 mumol/L nimodipine (voltage-gated calcium channel antagonist) (hereafter denoted as MCN) were administrated to hippocampal cultures, levels of [Ca(2+)](i) and cell death rate induced by KCN were partially reduced by 35.9% and 47.5% respectively. However, Gd(3+) (10 mumol/L) almost completely blocked KCN-mediated [Ca(2+)](i) elevation by 81.9% and reduced neuronal death by 88.8% in the presence of MCN. It is noteworthy that NGF, used in combination with MCN, inhibited KCN-induced [Ca(2+)](i) increase by 77.4% and reduced cell death by 87.1%. Only PLC inhibitor U73122 (10 mumol/L) abolished NGF effects. It is concluded that Gd(3+)-sensitive calcium influx, which is NMDA (glutamate receptor) and voltage-gated calcium channels-independent, is responsible for acute anoxic neuronal death. NGF can inhibit Gd(3+)-sensitive calcium influx and reduce anoxic neuronal death through activating PLC pathway.

AIM: To research the application analysis of trabeculectomy combined with phacoemulsification treating patients with primary angle-closed glaucoma ( PACG) and cataract.
METHODS: Seventy-three patients from September, 2011 to September, 2013, with primary angle- closed glaucoma and cataract, were selected in our research. Patients were all treated with trabeculectomy combined with ultrasonic emulsification, and the therapeutic effect was observed.
RESULTS: The effects of trabeculectomy combined with phacoemulsification treating primary angle-closed glaucoma and cataract in reducing intraocular pressure were better, and postoperative visual acuity results of the research objects were satisfactory. And it could deepen the central anterior chamber depth effectively. All the research objects had few complications.
CONCLUSION: Trabeculectomy combined with phacoemulsification treating primary angle - closed glaucoma and cataract can achieve a good result for reducing intraocular pressure, postoperative visual acuity recovery and correction, and the surgical success rate is high, with low complication rates. It is worthy of widely clinical application.

Objective To investigate the effects on one hand of training the other with isometric wrist extension and flexion training and its underlying mechanism.Methods Twenty healthy young girls were recruited and randomized into a training group and a control group with 10 subjects in each,using a random number table.The subjects in the training group were accepted isometric training of the wrist extensors and flexors on the right side once every other day,4 days a week for 6 weeks according to a pre-programmed protocol,while those in the control group had no intervention.Peak torque and surface electromyography (sEMG) were recorded and assessed before and after 6 weeks of training.Results In the training group,the average peak torque of right wrist flexion and extension were both significantly higher than before training (within-group comparison) and higher than in the control group (between-group comparison) after 6 weeks of training.The average peak torques of the left wrist in the training group in flexion and extension were (12.9±2.0) Nm and (6.4 ± 1.3) Nm after training,both significantly higher than before the training and stronger than the control group.In the sEMG traces during wrist extension,after training the ascend velocity of the right extensor carpi ulnaris and the integrated area of the left extensor carpi ulnaris in the training group were significantly smaller than before training and in the control group.Conclusion Unilateral isometric resistance training of the wrist muscles can transfer to the contralateral side,probably by altering muscle recruitment.

Objective To investigate the extent of cross-education between the pronation and supination muscles of the right and left forearms after unilateral isometric training.Methods Twenty healthy young girls were randomized into a training group and a control group using a random number table.The training group underwent isometric training of their right forearms for six weeks,while the control group continued ordinary life without exercises.Pronation peak torque (PPT) and supination peak torque (SPT) were assessed before and after the training for both groups.Results The differences in PPT or SPT between the training group and the control group were not significant before the training program.When the training had been completed,however,the average PPT and SPT on right side of those trained of course had increased significantly compared to before training or to the control group.More significantly,the average PPT and SPT on the left side in the training group were also significantly better than before training or in the control group after training.Conclusion Unilateral isometric training of the forearm pronation and supination muscles for six weeks can significantly increase muscle strength bilaterally,indicating good cross-education.

OBJECTIVE:To systematic review the relationship between CYP4F2 gene polymorphism and warfarin maintenance dose in Chinese population,and to provide evidence-based reference for clinical treatment. METHODS:Retrieved from Cochrane library,PubMed,EMBase,CJFD,VIP and Wanfang database,studies about the relationship between CYP4F2 gene polymorphism and warfarin maintenance dose in Chinese population were collected,and Meta-analysis was performed by using Rev Man 5.0 statis-tics software. RESULTS:A total of 13 studies were included,involving 2 958 patients. The type of gene was type TT,type CT and type CC. Results of Meta-analysis showed the relationship between 3 gene polymorphism and warfarin maintenance dose was type TT>type CT>type CC,and there were significant differences among groups. CONCLUSIONS:CYP4F2 gene polymorphism in Chinese population has significant correlation with warfarin maintenance dose. However,due to the limit of methodological quali-ty,large-scale and high quality studies are required for further validation of the conclusions.

Objective To investigate the effects of escitalopram (ESC)on the cognitive function,the expression of brain-derived neurotrophic factor (BDNF)in the hippocampus,the dendritic length and arborization and dendritic spines density of chronic cerebral ischemic rats.Methods Rats were randomly divided into sham-operation group,model group (permanent occlusion of bilateral common carotid arteries,2VO)and experimental group (treated with escitalopram at the dosage of 30 mg/kg·d).Rats were selected as study objects at week 1,2,4 and 8 after administration in each group.Their cognitive function was evaluated by the Morris water maze,the expression of BDNF protein was measured by Western blot,and dendritic morphology was studied by Golgi staining. Results In the Morris water maze test,the escape latency obviously extended in model group and experimental group compared with that in sham-operation group (P<0 .0 5 ),while the escape latency was shorter in experimental group than in model group (P<0.05).Compared with those in sham-operation group,the dendritic length and arborization and the density of dendritic spines in hippocampal CA1 significantly decreased in model group and experimental group (P<0 .0 5 ),while they increased significantly in experimental group compared with model group (P<0.05)by Golgi staining.Compared with sham-operation group,the expression of BDNF in the hippocampus of experimental group and model group significantly decreased (P<0 .0 5 ),but it increased significantly in experiment group compared with model group (P<0.05)by Western blot.Conclusion Escitalopram could significantly delay the progression of cognitive impairment induced by chronic cerebral ischemia in rats.The improvement of learning and memory may be related to the increased expression of BDNF.

OBJECTIVETo observe the intervention effects of electroacupuncture at "Yishu" (EX-B 3) on rats with type-2 diabetes mellitus (T2DM), so as to provide experiment references for acupuncture to treat T2DM.

METHODSAmong seventy male Wistar clean-grade rats, 8 rats were randomly selected into a control group; the rest rats were made T2DM model. Fifty-two rats which were successfully made T2DM model, according to randomized block method, were divided into a model group (10 rats), a medication group (10 rats), an electroacupuncture at "Shenshu" (BL 23) group (11 rats), an electroacupuncture at "Pishu" (BL 20) group (10 rats) and an electroacupuncture at "Yishu" (EX-B 3) group (11 rats). Seven days after successful establishment of model, the rats in the model group were fixed in the self-made rat bag without receiving any treatment; the rats in the medication group, according to body mass (10 mL/kg), were treated with intragastric administration of glimepiride; the rats in all the electroacupuncture groups were treated with electroacupuncture at "Shenshu" (BL 23), "Pishu" (BL 20) and "Yishu" (EX-B 3), respectively. The continuous wave was selected with a frequency of 15 Hz and a current intensity of 4 to 6 mA. The treatment was given 20 min per treatment, once a day, 5 treatments per week for continuous 4 weeks. Before the establishment of model and continuous 4 weeks after the intervention, blood samples were collected from rats' caudal vein, and fasting blood glucose (FBG) was measured with FBG device each week. After the last intervention, the rats were killed and hypothalamus, pituitary and adrenal gland were collected. The colorimetric method was applied to measure the contents of triglyceride (TG), cholesterol (TC), high-density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C); radioimmunoassay was used to test the contents of glycated serum protein (GSP), fasting insulin (FINS), corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortin (CORT).

RESULTSFour weeks after the intervention, except that the rat's body mass in the normal group continued to increase, body mass in the model group, medication group and each electroacupuncture group were significantly reduced (P<0.01, P<0.05). Compared with the model group, the FBG in the electroacupuncture at "Pishu" (BL 20) group and electroacupuncture at "Yishu" (EX-B 3) group were obviously reduced (P<0.05, P<0.01); FBG in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (both P<0.05). The contents of TG, HDL-C and LDL-C in the electroacupuncture at "Yishu" (EX-B 3) group were reduced (P<0.05, P<0.01), the content of TG was significantly lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (both P<0.05), the content of LDL-C was significantly lower than that in electroacupuncture at "Shenshu" (BL 23) group (P<0.05). Insulin sensitivity index (ISI) in the medication group, electroacupuncture at "Pishu" (BL 20) group and electroacupuncture at "Yishu (EX-B 3)" group were evidently increased (P<0.05, P<0.01); ISI in the medication group was lower than that in the electroacupuncture at "Yishu" (EX-B 3) group (P<0.05). The content of CRH in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (P<0.05, P<0.01); the content of CORT in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Pishu" (BL 20) group (both P<0.05).

CONCLUSIONElectroacupuncture at "Yishu" (EX-B 3) could reduce the level of CORT to improve the insulin resistance in rats with T2DM, improve insulin sensitivity index, regulate blood lipid metabolism and relieve the hyperactivity of the HPA axis.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; therapy ; Electroacupuncture ; Hormones ; metabolism ; Humans ; Hypothalamus ; metabolism ; Male ; Pituitary-Adrenal System ; metabolism ; Rats ; Rats, Wistar

Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.