To counteract the immune system in parasitic hosts, some viruses encode proteins to suppress the RNA interference (RNAi) effect. In this report, we established two RNAi systems to be easily observed with strong and obvious effect. The function of the P19 of tomato bushy stunt virus, which suppresses RNAi in mammal cells, was then studied using these two systems. Short hairpin RNAs targeting green fluorescence protein (pshRNA-GFP) and firefly luciferase (pshRNA-luc) were designed and inserted into a eukaryotic transcriptional vector pTZU6+1, respectively. The shRNA expressing vectors were co-transfected with plasmids containing the target gene with or without P19. The GFP expression level was assayed by fluorescence microscopy, Western blotting and RT-PCR. The luciferase expression level was analyzed by the dual-luciferase assay system. pshRNA designed in this study down-regulated the target gene specifically and efficiently, with a decrease of expression of both genes of about 70%, respectively. When P19 was introduced into the RNAi systems, the expression of both GFP and the luciferase were mostly recovered compared with the control groups. The RNAi systems of GFP and luciferase were constructed successfully, demonstrating that P19 of tomato bushy stunt virus has the ability to counteract the RNAi effect induced by shRNA in mammal cells.

To discuss the effective method of decreasing the postoperative recurrence rate of recurrent pterygium.
●METHODS:Totally 126 cases (126 eyes) with recurrent pterygium were randomly divided into A group (56 cases) and B group ( 70 cases ). Group A was treated by pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation, group B was treated by amniotic membrane transplantation. The followed-up time after surgery was 6-24mo.
●RESULTS:ln group A, postoperative 5-7d (average 5. 62± 1. 38d), cornea epithelium was repaired. ln group B, postoperative 7- 10d ( average 7. 38 ± 1. 12d), the corneal wound was healed. There was statistical significant difference between two groups (t = 4. 307,P<0. 05). Three cases recurrence were noted in A therapeutic group (56 cases), the recurrent rate was 5. 4%; Twelve cases recurrence were noted in B compared group (70 cases), the recurrent rate was 17. 1%. There was statistical significant difference between two groups(P<0. 05).
●CONCLUSlON: lt is suggested that pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation is effective in the treatment of recurrent pterygium.

Objective To study the relation of matrix metalloproteinase-9 in serum and tumor necrosis fac- tor-?in serum and amniotic fluid in predicting ehorioamnhionitis in patients with premature rupture of membranes (PROM).Methods The levels of MMP-9 in serum and TNF-?in serum and amniotie fluid were measured by ra- dioimmunoassay and ELISA in 67 cases with premature rupture of membranes as study group and 40 cases normal full-termed pregnant women as controls group.Results(1)The levels of TNF-?in amniotie fluid and MMP-9 in serum in study group were significantly higher than those in controls group(P0.05).(2)In study group,the levels of MMP-9 of serum in0.05).Conclusions The levels of TNF-?in amniotic fluid and MMP-9 in serum were valuable clinical indices for identification of chorioamnionitis in patients with PROM.The levels of MMP-9 in serum also could assess the time of rupture of membranes and the degree of ehorioamnionitis.

Objective To determine if Shenfu injectio(SFI)has any analeptic action in patients emerging from general anesthesia.Methods Eighty-six ASAⅠorⅡadult patients undergoing elective abdominal surgery under general anesthesia were randomly divided into 2 groups(n=43 each):SFI group and control group.The patients were premedicated with intramuscular phenobarbital 0.1g and atropine 0.5mg.Anesthesia was induced with propofol 2 mg?kg~(-1),fentanyl 4-5?g?kg~(-1) and vecuronium 0.1 mg?kg~(-1) and maintained with propofol infusion(2-4 mg?kg~(-1)?h~(-1)),0.5%-1.0% isoflurane inhalation and intermittent i.v.boluses of fentanyl and vecuronium.The patients were intubated and mechanically ventilated.The propofol infusion and isoflurane inhalation were stopped during skin closure.The patients were still unconscious and on mechanical ventilation at the end of surgery and transferred to PACU with a tube in trachea.As soon as the patients reached the PACU,SFI 1 ml?kg~(-1) in Ringer's solution 100 ml was infused over 10 min.In control group the patients received Ringer's solution 100 ml without SFI.The following times were recorded:(1)the time when the patients opened their eyes on command;(2)the time when mechanical ventilation was stopped;(3)the time when oxygen inhalation was stopped;(4)the extubation time;(5)the time of staying in PACU.Venous blood samples were taken before(T_0) and 5,15 and 45 min(T_(1,2,3))after SFI infusion for determination of plasma?-endorphin concentration.Results The awakening time,the mechanical ventilation time,oxygen inhalation time,extubation time and duration of PACU stay were significantly shorter in SFI group than in control group.There were no significant differences in MAP and HR after SFI between the two groups.The plasma?-endorphin concentration was significantly higher in group SFI than in control group.Conclusion Shenfu injectio can make patients emerging from general anesthesia faster.

OBJECTIVE: To investigate the application of power spectral entropy (PSE) in epileptic rats induced by penicillin, and to exploreits its value in predicting epileptic seizures. METHODS: Eighteen SD rats were randomly divided into 3 groups: a normal group, a control group and an epileptic group. Acute chemical models were made using penicillin microinjection in the right hippocampus of rats. Sprague-Dawley (SD) rats received continuous electroencephalographic (EEG) monitoring by deep department electrode. Changes of PSE of EEG signal in the epileptic group were analysed in the whole seizure process and compared with those in the normal group and the control group. RESULTS: In the whole seizure process of the rats induced by penicillin, PSE began to decline during the pre-ictal period and dropped sharply during the ictal period. PSE in the pre-ictal and ictal period declined apparently comparing with that in the non-ictal period. PSE in the epileptic group had significant differences during the ictal and pre-ictal period compared with that in the normal group and the control group (P<0.05). CONCLUSION Changes of PSE reveal the changes of the complex ictal EEG signals, and may be useful to predict epileptic seizures.
Animals ; Electroencephalography ; methods ; Entropy ; Epilepsy ; chemically induced ; diagnosis ; Male ; Nonlinear Dynamics ; Penicillins ; Predictive Value of Tests ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seizures ; chemically induced ; diagnosis ; Signal Processing, Computer-Assisted

OBJECTIVETo observe the effects and mechanisms of Pongamia pinnata root flavonoids (PRF) on the experimental gastric ulcer induced by acetic acid and to study the mechanism of PRF on the quality of ulcer healing.

METHODSThe models were established by acetic acid erosion, the quality of ulcer healing of PRF on the model of gastric ulcer were observed. The contents of epidermal growth factor (EGF) in serum were determined by radioimmunoassay. The expression of EGF and transforming growth factor-alpha (TGF-alpha) were detected by immunohistochemistry (SP).

RESULTSPRF significantly inhibited ulcerative formation induced by acetic acid (P < 0.05, P < 0.01). PRF could significantly increase the EGF and TGF-alpha (P < 0.05, P < 0.01) expression of para-ulcer mucosa tissue and improve the EGF contents in blood serum (P < 0.05, P < 0.01).

CONCLUSIONPRF increases the contents of EGF in serum and the expression of EGF and TGF-alpha in the tissue around gastric ulcer which might be one of possible mechanisms that PRF improves quality of ulcer healing.

Acetic Acid ; Animals ; Epidermal Growth Factor ; blood ; Female ; Flavonoids ; pharmacology ; Gastric Mucosa ; metabolism ; Male ; Millettia ; chemistry ; Plant Roots ; chemistry ; Rats ; Rats, Sprague-Dawley ; Stomach Ulcer ; chemically induced ; drug therapy ; metabolism ; Transforming Growth Factor alpha ; metabolism

OBJECTIVETo study the effect of HCV NS5A protein on HCV IRES-dependent translation in HepG2 cells.

METHODSHepG2 cells were co-transfected with a plasmid vector containing a bicistronic transcript carrying Renilla luciferase and firefly luciferase genes separated by HCV IRES sequences, and an expressing vector producing the NS5A protein. The luciferase activity and the mRNA of the luciferase gene were then detected. The NS5A expression was confirmed by fluorescence microscopy.

RESULTSHCV NS5A protein was detected in the cytoplasm of the HepG2 cells transfected with pcDNA-NS5A, and the luciferase activity was up-regulated in the presence of the HCV NS5A protein while the expression of luciferase mRNA showed no difference.

CONCLUSIONHCV NS5A protein can upregulate the HCV IRES activity and this effect is dose-dependent with NS5A.

Hep G2 Cells ; Hepacivirus ; genetics ; metabolism ; Humans ; Plasmids ; Protein Biosynthesis ; Protein Structure, Secondary ; Ribosomes ; metabolism ; Transfection ; Viral Nonstructural Proteins ; metabolism

BACKGROUND: The proliferation of peripheral blood stem cells among peripheral blood mononuclear cells (PBMCs) invitro remains unclear. There is no optimal marker for tracing PBMCs transplanted in vivo.OBJECTIVE: To observe the degree of PBMC proliferation in stem cell medium by EdU labeling and to explore thefeasibility of EdU-labeled peripheral blood stem cells.METHODS: New Zealand rabbit PBMCs were isolated and cultured for 1 to 5 days in stem cell medium supplementedwith EdU. The cells were observed and counted at 0, 1, 2, 3, 4 and 5 days in culture. The cells were harvested at eachtime point and stained with EdU fluorescent reagents. Then, confocal microscopy and flow cytometry were used to detectEdU-labeled cells.RESULTS AND CONCLUSION: (1) Freshly isolated rabbit PBMCs were rounded and showed clear outline. After 1 dayculture, most of the cells were suspended in the medium, spherical or round. There were also a few cell clusters andadherent cells scattered in a triangle or polygon shape; after 2 days culture, more cell debris were observed, and mostcells were round; when cultured for 3-5 days, increased cell debris, smaller cell mass and decreased cell densitysignificantly were observed. (2) With the prolongation of culture time, the cell count decreased gradually. (3) Whencultured for 1 day, EdU labeled cells in red were scattered. The number of cells marked with EdU red label increasedsignificantly at day 2 and remained unchanged after 3 days of culture. At 5 days of culture, the number of red cellsmarkedly decreased; the highest positive rate of EdU-labeled cells was (2.38±0.10)% at 2 days after culture. To conclude,these results showed that the proportion of proliferating cells in rabbit PBMCs was very low. EdU is capable of labelingproliferative cells among PBMCs.

To investigate the effects of conducting electrodes implanted into the penicillin-induced epilepsy foci in rats on seizures, 30 male SD rats were randomly divided into experimental group (EG) and control group (CG). Afer penicillin(PNC) administration, metal electrode was implanted into the epilepsy focus in EG. Behavior evaluations were given after PNC injection in both groups and electroencephalogram recordings were taken respectively. Immunohistochemical methods were used to test the expressions of P38 and Calcium channel a 1A subunit (CACNA1A) at the serial time-point. We found that EG showed reduction of epileptic discharge frequency than did CG at time-point 3 h and 6 h, and that P38 expressions of EG were lower than those of CG in the injected side, and the expressions of Calcium channel a 1A subunit (CACNA1A) were downregulated in EG at time-point 6 h and 24 h. This evidence suggests that the metal electrode implanted into the penicillin-induced epilepsy focus may reduce the epileptic discharges and downregulate the expressions of P38 and CACNA1A. This new idea might be used in antiepileptic field in future.
Animals ; Calcium Channels ; metabolism ; Down-Regulation ; Electrodes, Implanted ; Electroencephalography ; Epilepsy ; chemically induced ; therapy ; Male ; Penicillins ; Rats ; Rats, Sprague-Dawley ; p38 Mitogen-Activated Protein Kinases ; metabolism

Teaching rounds are an important part of hospitals at all levels to carry out clinical teaching work and improve medical quality. Taken anesthesia-related knowledge as the center, based on the patient's pathophysiology and anesthesia physiological changes, our department asked the core questions such as the impact of the operation method and anesthesia plan, then students answered, and finally closed the loop after the case. This process is called problem-based learning (PBL) teaching rounds. It is found that the teaching rounds can help improve the ability of anesthesiologists to solve problems, learn independently, and learn throughout their lives, thereby improving the teaching quality of standardized residency training of anesthesiology.