BACKGROUND:The HOX gene family is a highly conserved transcription factor family,which affects the formation of basic axis and secondary axis during embryonic development,at the same time,it plays a pivotal role in the development of the central nervous system,axial skeleton,stomach intestine,urogenital system and external genitalia.OBJECTIVE:To investigate the interactional protein of HOXD13 in human embryo foot development at 14 weeks pregnancy.METHODS:Fetal foot tissues were harvested and protein extraction was performed using protein extraction kit,6 μg HOXD13 antibody was added for immunOprecipitatjOn experiment,which aims to obtain HOXD13 interactional proteins,while the non-specific IgG served as a negative control Subsequent to Bradford staining,the sediment protein bands were cut for mass spectrometry.RESULTS AND CONCLUSION:We obtained an interactional protein of HOXD13 in the human embryo foot at 14 weeks pregnancy,at the relative molecular mass of approximately 60 000.The protein was identified as zinc finger protein 548 using mass spectrum.HOXD13 may play a very important role in transcriptional control by HOXD13-zinc finger protein 548 protein complex in the human embryo foot at 14 weeks pregnancy.

BACKGROUND: Preliminary study shows that the FHL1 gene expression is down-regulated in muscle tissue of patients with clubfoot, and the gel retardation experiments have verified HOXD13 and FHL1 gene promoter region transcription factor predicted binding sites in vitro, but the experimental results are not enough to truly reflect in vivo transcriptional regulatory protein and DNA binding conditions.OBJECTIVE: Western-blot technique is utilized to further validate FHL1 and HOXD13 gene expression in protein levels in muscle tissue of patients with congenital clubfoot, the in vivo HOXD13 and predicted binding sites in embryonic foot development was verified using chromatin immunoprecipitation technology.METHODS: Muscle tissues were samples from 15 children with congenital clubfoot, in the Department of Pediatric Surgery, at the Second Affiliated Clinical Hospital of China Medical University, 3 copies of normal children foot muscle tissues at the same age were provided by the Forensic Medicine College of China Medical University, 1 case of aborted embryo at pregnancy 13 weeks were offered by Department of Obstetrics and Gynecology, at the Second Affiliated Clinical Hospital of China Medical University. All specimens used are given informed consents by the patients and their families. Western-blot method was applied to detect the expressions of HOXD13 and FHL1 in foot muscle tissue of 15 patients of congenital clubfoot and 3 normal children at the same age; The FHL1 gene upper stream HOXD13 binding sites was predicted using software, the interaction of HOXD13and FHL1 during embryonic development was verified with chromatin immunoprecipitation experiment, brain tissue without HOXD13 protein expression served as controls.RESULTS AND CONCLUSION: Compared with normal children foot muscle tissues, both HOXD13 gene (5/15) and FHL1 (7/15)were down-regulated in 15 patients of congenital clubfoot. An enrichment of the predicted HOXD13 binding site was observed in the precipitated human embryo foot tissues chromatin. No enrichment of the predicted site containing sequence was observed in the control brain chromatin; additionally, there was no enrichment of the control sequence. This study further verifies that, the FHL1 and HOXD13 gene expression are down-regulated in the foot muscle tissue of congenital clubfoot children patients; during development of human embryos, HOXD13 protein can bind with FHL1 promoter region binding sites to play its role in transcriptional regulation. It is indicated that in the human foot embryonic development, the down-regulation of HOXD13expression may result in reduced expression levels of FHL1, thereby affecting the foot muscle growth and differentiation, leading to clubfoot deformity occurring.

Objective To evaluate the efficiency and safety of amiodarone in patients with acute myocardial infarction (AMI) complicated ventricular tachyarrhythmia (CVT). Methods 106 CVT patients of AMI with stable haemodynamics was randomized into trial group (53 cases) and control group (3 cases). Based on routine therapy, the trial group was intravenously given amiodarone. Electrical cardioversion is necessary if the haemodynamics turns to unstable. Intravenous amiodarone will be used for at least 24 hours to maintain sinus rhythm. The control group was administrated intravenous lidocainein. If the patients made no response to lidocainein, given amiodarone as substitute. Electrical cardioversion is necessary when the haemodynamics turns to unstable and lidocainein was followed for at least 24 hours after successful cardioversion to maintain sinus rhythm. The therapeutic effects, cardiac function and the changes of arrhythmia were compared between the two groups. Results The incidence of angina pectoris, consumption of nitrates were decreased in trial group when compared with that in control group, whereas the ejection fraction, left ventricle fast filling interval and the mitral valve peak velocity of blood flow during left atrium contraction(E/A) all were higher than that in control group (all P<0.01). The total effective rate in trial group was higher than that in control group (75.5% vs 62.3%, P<0.01), especially the ventricular tachycardia control rate is significantly higher than control group (86.7% vs 50.0%,P<0.01). Conclusion Intravenous injection of amiodarone efficaciously control the complicated ventricular tachy-arrhythmia in patients with acute myocardial infarction as well as to improve the cardiac function.

It had fertile countryside land,rich products,dense population and creeks and lakes in ancient Hangzhou and its neighbour area,where there is a good place for planting medicinal materials.It had long history of planting medicinal materials with abundant output,pure and good nature,and much varieties;among which,some famous and costly medicinal materials,such as "eight Zhe's" and "eighteen Hangs",were not only adopted broadly by court,royalty and doctors,but purchased by many businessmen and sold far to abroad as well;they seemed to be in the leading place in China.

Objective To study the effects of Multi-Walled Carbon Nanotube with different doses on the cellular proliferation of human embryo kidney 293 cells.Methods The cultured human embryo kidney cell line 293 was seeded into 96-well plates and various concentrations of Multi-Walled Carbon Nanotube were added into different groups of culture.Methyl thiazolyl tetrazolium(MTT)assay were applied to detect the cellular viabilities after being incubated with Multi-Walled Carbon Nanotube for 48 hours and the results were calculated with specific static software to protract the cell viability curve.Results The Multi-Walled Carbon Nanotube had different effects of inhibiting the multiplies of 293 cells depending on its concentrations.In addition,when the concentration of Multi-Walled Carbon Nanotube reached over 0.5ug/ml,the inhibition became significantly.Conclusions Multi-Walled Carbon Nanotube can penetrate the cell membrane of 293 cells to influence the activities of cells and the ability of proliferation will be decreased significantly when the concentration came to some degree.Therefore,the safety dosage of the Multi-Walled Carbon Nanotube on human normal embryo kidney 293 cells were estimated in this research.

OBJECTIVE:To establish a method for determining plasma concentrations of tanshinone ⅡA and use it for pharma-cokinetics study of tanshinone ⅡA in Beagle dogs. METHODS:HPLC was performed on the column of Phenomenex Luna C18, with the mobile phase of water(5 mmol/L of ammonium acetate+0.05% H3PO4)-acetonitrile at flow rate of 1.0 ml/min;the detec-tion wavelength was 270 nm,column temperature was 40 ℃ and the volume was 20 μl. 9 Beagle dogs were randomly divided into tanshinone ⅡA low,medium and high dose groups(2,4 and 8 mg/kg),blood was respectively taken from forelimb venous blood to determine the plasma concentration before and after 2,5,10,15,20,30,45,60,75,90 and 120 min of iv administration. DAS 2.0 software was used to calculate the pharmacokinetic parameters. RESULTS:The linear range of tanshinoneⅡA was 0.097 5-12.50 μg/ml (r=0.999 8);the RSDs of precision and stability tests were all less than 10%;the method recovery was 100.4%-107.2%,and extraction recovery was 90.2%-92.3%. The t1/2αin tanshinoneⅡA low,medium and high dose groups were respective-ly(1.01±0.27),(1.03±0.46)and(1.51±0.65)min;t1/2β were respectively(16.25±4.78),(22.42±9.32)and(24.45±12.02) min;AUC0-120 min were respectively(150.88±45.25),(305.44±92.55)and(643.67±178.27)μg·min/ml;and CL were respective-ly(12.01±4.36),(12.78±5.06)and(12.17±5.41)ml/(min·kg). CONCLUSIONS:The precision,stability and recovery of the method are all in line with the determination requirements of biological samples,and tanshinone ⅡA showed a good linear relation-ship with dose in Beagle dogs in AUC0-120 min.

OBJECTIVE:To increase the accuracy of blood concentration monitoring results of tacrolimus.METHODS:Blood concentrations of tacrolimus(60points for high and low quality control,respectively)were determined by enzyme-linked immunosorbent assay and the quality control graphs were drawn.The whole monitoring course underwent both prospective and retrospective quality control.RESULTS:The quality control monitoring results could be evaluated from the quality control graphs.CONCLUSION:Quality control can effectively increase the accuracy of blood concentration monitoring results of tacrolimus,which can also prevent and rectify systemic error.

Objective To study whether tissue injury could be increased by double shock wave while it improves the efficiency of lithotripsy. Methods A series of experiment study on the tissue injury in vivo pigs with double and single shock wave as well as electromagnetic shock wave has been carried out. Results Under the conditions of 8 kV and 1 000 times concussion, the trauma range of kidney is 18 mm?15 mm?3 mm with single shock wave,and 14 mm?11 mm?2 mm with double shock wave,the trauma range of ureter is 42 mm with single shock wave and 30 mm with double shock wave.Under the same energy and concussion,the injury focus induced by double shock wave is small than that induced by single shock wave.Microstructure pathology shows that the characteristics of focus on renal artery,vein,glumeruli,renal tubules and ureter are similar. Conclusions Compared with the focus induced by single or by electromagnetic shock wave,the focus on the tissue of kidney and ureter induced by the double shock wave is smaller.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) with multiple differentiation potential can be induced into osteogenic or adipogenic differentiation under certain conditions. OBJECTIVE: To review the related factors regulating the adipo-osteogenic differentiation of BMSCs. METHODS: A computer-based search of CNKI and PubMed databases was performed for literature concerning the related factors regulating the adipo-osteogenic differentiation of BMSCs published from January 2006 to August 2016. The search terms were bone marrow mesenchymal stem cells, osteogenic differentiation, adipocyte differentiation in Chinese and English, respectively. RESULTS AND CONCLUSION: Signaling pathways, transcription factors, and smal molecule compounds that are interacted are key factors in the regulation of BMSCs differentiation, so the techniques to intervene BMSCs differentiation based on these key molecules may correct bone or fat abnormality and can be applied to tissue engineering and regenerative medicine in the future. Additionally, the biological clock is also one of the most important factors for adipo-osteogenic differentiation of BMSCs by regulating signaling pathways or transcription factors.

BACKGROUND:The third molar extraction may lead to different degrees of soft tissue damage. Ultrasound and ultrashort waves belong to hyperthermia, which can increase tissue temperature and improve the therapeutic efficacy of soft tissue stretch. OBJECTIVE:To observe the effects of soft tissue stretch combined with ultrasound and ultrashort wave therapy for patients who were recently diagnosed with trismus and pain folowing third molar removal. METHODS:Sixteen patients with trismus and pain after third molar removal were randomly divided into two groups: patients in stretch group received soft tissue stretch with ultrasound and ultrashort wave therapy, while those in control group received only ultrasound and ultrashort wave therapy. Patients received the treatment five times per week, totaly for 2 weeks. RESULTS AND CONCLUSION:It was found that after treatment pain was significantly reduced at the temporomandibular joint, and the range of maximum active mouth opening increased significantly. The improvement in mouth opening and temporomandibular joint function was better in the stretch group than the control group. These findings indicate that ultrasound and ultrashort wave therapy can improve symptoms of trismus and pain folowing third molar removal, and the combination of ultrasound and ultrashort wave therapy and soft tissue stretch therapy can achieve more effective results.