The zebra fish model, as an integral animal model, features small volume, high throughput, low cost, short cycle and reliable experimental results, thus has been widely used in medical studies. Traditional Chinese medicines (TCM) constitute a complex system, their active ingredients and action mechanisms are among study hotspots in during the development of modern TCMs. Along with the constant improvement of advanced technologies and methods, zebra fishes have been increasingly applied in studies on TCMs and shown advantages in active screening, and toxicity and metabolism studies. In this paper, TCM studies by using zebra fishes in recent years are summarized to provide new ideas and methods for basic studies on TCMs.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Medicine, Chinese Traditional ; Models, Animal ; Zebrafish ; genetics ; growth & development ; metabolism

Objective To screen the valid plasmid targeted toGCN5 among the constructed recombinant plasmids;and to explore the effects of histone acetylizad modification in regulating MSCs differentiation.Methods Exstract the constructed plasmids and transfect them into MSCs induced by 5-aza.For 24 h,observe MSCs;detect the transfect efficiency by flow cytometry;detect expression of protein GCN5 by Western-blot;detect the HAT activity by ELISA.Results Transfect efficiency was more than 20%;Expression of protein GCN5 and HAT activity had no difference in group ZJ1 and ZJ2,but had a significant difference to that in group ZJ3.HAT activity of experiment group was significantly lower than that of control groups.Conclusion The inhibition state of histone acetylation results in plasmid ZJ3,it can inhibit the differentiation process of MSCs.The results provide data for the clinical application of MSCs transplantation.

Objective To study the expression of E-cadherin( E-cad), p-catenin(β-cat) in labial salivary glands of patients with primary Sj(o)gren's syndrome (pSS) in order to explore their role in pathogenesis. Methods Biopsies of labial salivary glands were obtained from 52 patients with pSS and 30 healthy controls. The immunohistochemical staining was used to detect the expression of E-cad and β-cat. Anti-SSA and anti-SSB antibodies were measured. Semi-quantitative analysis was performed by image analysis software. Ultra-structural changes was used by electron-microscopic techniques. Results ① The area of expression, optical intensity and the accumulated optical intensity of the E-cad group [(2513±1086) μm2, 0.212±0.041, 566 ±297 ] were lower than normal controls. The expression level was reduced as the increase of lymphocyte infiltration focus. ② The area of expression, the optical density and the accumulated optical density of the β- cat group [(12 324±7883) μm2, 0.113±0.031, 566±297] was lower than those of the control group. The expression level was reduced as the increase of the lymphocyte infiltration focus. ③ The E-cad expression and the p-cat expression was positively correlated in the labial gland of patients with pSS. ④ Howev-er, there was difference in the expression of E -cad and β -cat between patients with positive SSA and negative SSA antibodies. Conclusion In salivary samples, the expression of both E-cad and p-cat in patients with pSS is lower than those of the controls. Anti-SSA/SSB antibodies are important parameters of pSS and they may be involved in the pathogenesis of pSS.

This research uses six Agrobacterium rhizogenes R1601, R15384, R1000, A4, R1025 and R1 to infect silymarin explants to induce hairy roots and silibin. All of the six A. rhizogenes can induce Silybum marianum to generate hairy roots and the A. rhizogene A4 shows comparatively high infection on the plant. This research determines the condition to induce silymarin hairy roots by the factors of infection time, pre-culturing, co-culturing and pH value. The fact that MS liquid medium fits the proliferation of silymarin hairy roots is determined. Through PCR molecular identification, it can be seen that the DNA plasmids in the A. rhizogenes are successfully integrated into the genome of transformed roots. Using liquid chromatography, it is determined that the silibin content in silymarin hairy roots is 2.5 times that in the plant In this research, the silymarin hairy roots culturing system is established, which lays a foundation for the study of culturing silymarin hairy roots and producing silibin.
Agrobacterium ; genetics ; physiology ; Cell Culture Techniques ; methods ; Milk Thistle ; chemistry ; genetics ; growth & development ; microbiology ; Plant Roots ; chemistry ; genetics ; growth & development ; microbiology ; Silymarin ; analysis ; Transformation, Genetic

Objective To assess the function of motor nerve fiber in patients with diabetic peripheral neuropathy (DPN) by single-fiber conduction studies.Methods According to the diagnostic standard of DPN issued in Toronto meeting in 2009,on the basis of the result of peroneal nerve conventional conduction study,a total of 65 patients with DPN in the Department of Endocrinology and the Department of Neurology of Tianjin Third Central Hospital from October 2012 to October 2013 were enrolled into the study,from whom 33 had abnormal sensory conduction (sensory-diabetic peripheral neuropathy group,S-DPN group),32 had abnormal sensory motor conduction (sensory motor-diabetic peripheral neuropathy group,SM-DPN group).Single-fiber conduction velocity (SF-CV) and single-fiber distal motor latency (SF-DML)were detected in all subjects.The obtained results were compared with the data from 34 healthy volunteers (control group).The relationship of SF-CV,SF-DML and the duration of diabetes mellitus,fasting glucose,HbA1 c was also studied in DPN patients.Results The SF-CV ((43.1 ± 3.6) m/s) was decreased in S-DPN group compared with control group ((47.5 ± 3.3) m/s,t =5.077,P ＜ 0.01).There were no significant differences in SF-DML ((3.6 ± 0.7) ms),motor nerve conduction velocity (MCV (49.5 ± 2.6)m/s) and DML ((3.4 ± 0.6) ms) in S-DPN group compared with that of control group ((3.4 ± 0.5) ms,(50.9 ± 3.5) m/s,(3.2 ± 0.5) ms,respectively).SM-DPN group had lower SF-CV ((35.2 ± 3.6)m/s,t =9.119,14.219),MCV ((40.9 ± 3.2) m/s,t =11.131,13.025) and increased SF-DML ((4.5±0.7) ms,t=5.692,7.231),DML ((4.2 ±0.7) ms,t=5.561,6.975) compared with the other two groups (P ＜0.01).SF-CV in DPN patients was negatively related to the diabetic duration (r =-0.340,P =0.006),while SF-DML had no correlation with duration of DM,fasting blood glucose and HbAlc.Conclusions Detection of SF-CV is easy to find early motor nerve dysfunction in DPN patients.SF-CV is decreased with the increasing duration of diabetes.

Objective To observe the response of the guiding vane to axial flow pump by testing two types of miniature axial flow pump,which have been developed by our group in recent years.Methods Two types of axial flow pump were tested in a mock circulation apparatus.Results The outputs of the two types of miniature axial flow pump all attain 5L/min against the pressure of 13.332kPa,which is enough for left ventricular assistance.The outputs are similarly related to their pre-load and after-load pressure.In this study,the guiding vane does not produce a significant affection on the hydrodynamic characteristic of the axial follow pump.Conclusion The guiding vane may be neglected in the design to improve the blood compatibility of a axial flow pump heart assist device.

Objective To compare diagnostic performance of ultrasound and radionuclide imaging in diagnosis of Meckel's diverticulum.Methods Totally 46 children suspected with Meckel's diverticulum were enrolled.Ultrasound,radionuclide imaging data were analyzed and compared with pathology.Results In 46 children suspected with Meckel's diverticulum,38 cases were confirmed by operation.Thirty-three cases of 38 were diagnosed Meckel's diverticulum by ultrasound,5 cases of 38 were false negative,there was no false positive case.Radionuclide imaging was positive in 24 cases of 46,false positive in 4 eases and false negative in 18 cases.The ultrasound diagnostic accuracy rate was 89.13％ (41/46),sensitivity was 86.84％ (33/38),specificity was 100％ (8/8).Diagnostic accuracy rate of radionuclide imaging was 52.17％ (24/46),sensitivity was 52.63 ％ (20/38) and specificity was 50.00 ％ (4/8).The sensitivity of ultrasound and radionuclide imaging in diagnosis of Meckel's diverticulum had significant difference (P＜0.01).Conclusion Ultrasound in diagnosis of Meckel's diverticulum has advantages of non-invasive,no radiation,acceptable price and high sensitivity.

Objective To observe the clinical effects of pars plana vitrectomy (PPV) combined with internal limiting membrane peeling (ILMP) for macular hole (MH) and macular retinoschisis in high myopic eyes,and to analyze factors affecting the MH closure rate.Methods This is a retrospective case study.21 high myopic patients (22 eyes) with MH and macular retinoschisis were enrolled in this study.All eyes were examined for best corrected visual acuity (BCVA),intraocular pressure,slit lamp microscope,indirect ophthalmoscope,A and/or B-scan ultrasound,optical coherence tomography and visual electrophysiological examination.The BCVA was ranged from finger counting to 0.2.The axial length (AL) was ranged from 26.00 to 31.00 mm,with an average of 27.47 mm.Among 22 eyes,AL was between 26.00 mm to 27.00 mm in 9 eyes,27.10 mm to 28.00 mm in 5 eyes,28.10 mm to 29.00 mm in 3 eyes,29.10 mm to 30.00 mm in 3 eyes,and longer than 30.00 mm in 2 eyes.The diameter of MH was ranged from 227 μm to 597 μm and with an average of 432 μm.Among them,the minimum diameter was between 200 μm to 400 μm in 4 eyes,401 μm to 450 μm in 13 eyes,451 μm to 500 μm in 3 eyes,501 μm to 600 μm in 2 eyes.All the eyes were treated with PPV combined with ILMP surgery.The average follow-up time was 17 months after surgery.The efficacy was determined at the final follow up,including the MH closure,the state of macular retinoschisis and the BCVA.MH closure rate with different MH diameters and different AL were compared and analyzed.Results During the final followup,MH were fully closed in 17 eyes (77.3％),bridge-closed in 4 eyes (18.2％) and not closed in 1 eye (4.5％).Retinoschisis was resolved in 19 eyes (86.4％),partially resolved in 2 eyes (9.1％) and not changed in 1 eye (4.4％).MH with smaller diameter had higher MH closure rate (x2=12.036,P=0.032).MH with longer AL had lower MH closure rate (x2=16.095,P=0.003).The final BCVA was ranged from finger counting to 0.25.Among 22 eyes,BCVA or metamorphopsia were improved in 9 eyes (40.9％),stable in 8 eyes (36.4％).BCVA was reduced and metamorphopsia was more severe in 5 eyes (22.7％).Conclusions PPV combined with ILMP is a safe and effective surgical treatment for MH (with minimum diameter ≤600 μm) and macular retinoschisis in high myopic eyes.After surgery,MH was closed and retinoschisis was resolved in most patients.The major factors affect the MH closure were the minimum diameter of MH and AL.

Objective To investigate the protective effect of matrine on lung injury associated with single lung ventilation during thorac-ic surgery,and to explore and consummate the prevention and control measures of single lung ventilation related lung injury.Methods To-tally 97 cases of non small cell lung cancer patients were randomly divided into the observation group ( 50 cases ) and the control group (47 cases) .The two groups of patients were given the same way of anesthesia.Patients of the observation group received intravenous drip of 2 mL matrine injection which were dissolved in 100 mL saline solution 30 min before anesthesia, while patients of the control group were merely given 100 mL saline solution 30 min before anesthesia.The pulmonary shunt fraction( Qs/Qt) ,xanthine oxidase( XOD) ,myeloperoxi-dase(MPO),superoxide dismutase(SOD) and nitric oxide(NO) of the following points in time were compared:before anesthesia induction (T0),the instant of OLV (T1),60 minutes after OLV(T2),120 minutes after OLV(T3),after lung inflation (T4),and 24 hours after opera-tion ( T5) .Results At the time of T1 to T4,pulmonary shunt fraction of the two groups were both significantly higher than that at T0 with sig-nificant difference ( P<0.05) ,but there was no statistical difference in terms of intra-group comparison at different time points ( P>0.05) .The PMN counts of the two groups at the time of T2 to T5 were significantly higher than that of T0 with significant difference (P<0.05),and the PMN counts at the time of T2 to T5 in the control group were significantly higher than that in the observation group with significant difference (P<0.05).The levels of serum XOD,MPO,and SOD at T2 to T4 in both of the two groups were significantly higher than that at T0 with signif-icant difference (P<0.05),and the serum levels of XOD,MPO and SOD at T2 to T4 in the control group were significantly higher than those in the observation group with significant difference (P<0.05).The levels of serum NO at T2 to T4 in both of the two groups were significantly higher than that at T0 with significant difference (P<0.05),and it was significantly lower than that in the observation group with significant difference (P<0.05).Conclusion The matrine pretreatment of lung injury in the patients with single lung ventilation has a protective effect, which can reduce the levels of oxidative stress and promote the NO release in patients by reducing PMN,XOD and MPO levels.

Objective: To study influence of acute hypoxia on the current of voltage-gated potassium channel (IK) in pulmonary artery smooth muscle cells (PASMC) of rats. Methods: A total of 20 male SD rats were randomly and equally divided into normoxic control group and acute hypoxia group. The rats in acute hypoxia group were kept in hypoxic chamber for 8 h before experiment. Whole cell patch-clamp technique was used to record IK in PASMC. Results: Acute hypoxia significantly decreased the IK density in PASMC of rats. During -60mV to -10mV of resting membrane potential（RMP）, acute hypoxia did not significantly decrease peak IK density in PASMC of rats, P>0.05; At 0 mV, acute hypoxia significantly decreased the peak IK density in PASMC [from（38.1 ± 5.2） pA / pF decreased to（9.82 ± 2.1） pA / pF ,P<0.05], then along with RMP increase in PASMC, the decreasing amplitude of peak IK density in PASMC gradually increased（P<0.05）; From + 30 mV to+ 60 mV, the decreasing amplitude of peak IK density in PASMC further significantly increased（P<0.01）; At + 60 mV the peak IK density decreased from（38.1 ± 5.2） pA / pF to（9.82 ± 2.1） pA / pF , and the decreasing amplitude reached (46.8±3.3)%. Conclusion: Acute hypoxia can decrease Kv current in PASMC of rats, leading to hypoxic pulmonary vasoconstriction.