Objective To study the optimum parameters of the supercritical CO2 fluid extraction of Polyrhachis vicina Roger and chemical constituents of ant oil. Methods The experiments were designed with the uniform design. Three factors are extraction pressure, temperature and time. GC-MS was applied for analyzising. Results The optimum condition was obtained:the extraction pressure is 36 MPa, the temperature of extraction is 40 ℃ and extraction time is 50min. 45 components were identified from 60 chromatographic peaks. The main chemical constituents were (Z)-9-octadecenoicacid (48.278%), hexadecanoicacid (17.574%), (Z)-9-hexadecenoi (4.912%), cholesterol (2.957%). There are some linear paraffin in ant oil. Conclusion This method can extract the ant oil from Polyrhachis vicina Roger quickly and efficiently. This experiment provide foundation for exploitation and utilization of ant resource.

Objective To dynamically monitor the effect of Cinepazide maleate (CM) on ischemic region of the brain and to elucidate the neuroprotection of CM on acute cerebral ischemic rat model and related mechanisms.Methods The rat model of acute brain ischemia-reperfusion was established with tighting threads.They were randomly divided into ischemia-reperfusion group and CM treatment group(n=8 in each group).Rats in CM treatment group were given 3 mg / kg CM by caudal intravenous injection immediately after brain ischemia-reperfusion.T2 weighted imaging(T2WI) and proton magnetic resonance spectroscopy (1HMRS) were performed before operation and at 6 hours,24 hours and 3 days after reperfusion.Results At 6 h,24 h and 3 d after operation there was no difference for areas of cerebral ischemia on T2WI between 2 groups.The values of N-acetylaspartate(NAA) (Phosphocreatine(PCr) +Creartine(Cr)) in ischemia-reperfusion group rats were (1.53±0.20),(0.50 ±0.17),(0.44±0.13),(0.40±0.10) before operation and at 6 h,24 h,3 d after operation,respectively.And in CM treatment group the values of NAA/(PCr+Cr) were (1.44±0.22),(0.68±0.13),(0.61±0.10),(0.53±0.09) at the same time points.The values of lactate(Lac)/(PCr+Cr) in ischemia-reperfusion group were (0.03±0.01),(1.10 ±0.28),(1.30± 0.23),(1.23± 0.19) before operation and at 6 h,24 h,3 d after operation,respectively.And in CM treatment group the values of Lac/(PCr+Cr) were (0.02±0.01),(0.85±0.25),(0.99±0.20),(0.90±0.15) at the same time points.The rats in ischemia-reperfusion group had lower value of NAA/(PCr+Cr) (P＜0.01) and higher value of Lac/(PCr+Cr) at all time points after operation than those before operation.Compared with rats in ischemia-reperfusion group,the value of NAA/(PCr+Cr) increased(P＜0.05) and the value of Lac/(PCr+Cr) decreased(P＜0.05) decreased significantly for CM treatment group rats.Conclusion CM treatment can decrease the intracellular accumulation of lactic acid and reduce neuronal necrosis in acute brain ischemia-reperfusion rats.

ABSTRACT:The aim of the present study was to investigate the inhibitory effects of TLR7 on Mycobacterium tuberculosis . TLR7 on infected RAW264 .7 cells was activated by chemical synthesis of TLR7 activation motif ssRNA .Activated RAW264 .7 cells were inoculated with Mycobacterium tuberculosis ,quantitative PCR method was applied to detect the phagocytosis rate of cell to bacteria at different time after infection .Cytokine production was measured by ELISA from cell supernatant .Cells were cultured on Roche medium and counted after sterile cracked with TritonX‐100 and diluted with PBS .Scanning electronic micro‐scope ( SEM ) was applied to detect the morphological changes of cells treated with TLR7 activation motif ssRNA .The highest phagocytosis rate of bacteria of RAW264 .7 cells was at 3 hours post infection (P>0 .05) .Compared with that of the control group ,treatment after 36 hours intracellular bacterial quantity in ssRNA treated group was lower (P<0 .05) ,levels of IL‐12 (P<0 .05) and IL‐4 (P<0 .05) were increased .For treatment after 48 hours ,level of IL‐4 (P<0 .05) was decreased ,and TNF‐α (P<0 .05) was increased .For treatment after 3 hours ,cell morphology of the ssRNA group was obviously better than the control group and appeared lots of phagosomes .Results suggested that TLR7 could enhance macrophages in killing Myco‐bacterium tuberculosis by forming phagosomes and regulating cytokines production ,and TLR7 activation motif ssRNA could be used in the treatment of tuberculosis .

To study the in situ intestinal absorption kinetics and compatibility influence of peimine and peiminine in rats, the absorption of peimine and peiminine in small intestine (duodenum, jejunum and ileum) and colon of rats was investigated using in situ single-pass perfusion method and the drug content was measured by HPLC-ELSD. Perfusion rate, pH, concentration of drug, gender and bile duct ligation can significantly affect the absorption of peimine and peiminine, the Ka, and Papp values in the condition of pH 6.8 and pH 7.4 had significant difference (P<0.01), as drug concentration irlcreased, the absorption parameters of peimine and peiminine decreased, Ka and Papp between low concentrations and middle concentrations was significant difference (P<0.01). Verapamil can not affect Ka and Papp of peimine and peiminine which are in the extract (P> 0.05). Bitter almonds and licorice can significantly reduce the absorption of peimine and peiminine with the usual dose (P<0.01), extracted separately and together had no significant difference on Ka and Papp (P> 0.05). Experimental results show that the absorption features of peimine and peiminine are basically the same, both of them could be absorbed at all segments of the intestine in rats and had no special absorption window, and with significant differences between male and female individuals. The absorption of peimine and peiminine complies with the active transport and facilitated diffusion in the general intestinal segments. Bitter almond and licorice can reduce the intestinal absorption rate ofpeimine and peiminine.

Objective To evaluate the clinic effect of leaflet enlargement with autologous pericardium in repairing mitral valve disease and to describe the technique and discuss its indications. Methods Between July 2004 and June 2008, 45 pa-tients with isolated mitral valve disease, included stenosis in 10 and regurgitation in 35. The causes were congenital heart dis-ease in 8, rheumatic in 21, degenerative in 7 and endecarditis in 9. The procedures were: posterior leaflet enlargement with autologuus pericardium in 14, anterior leaflet enlargement in 8, both anterior and posterior leaflet enlargement in 23. In addi-tion, eye to eye technique was in 12, artificial chordal in 12, chordal transfer in 6, papillary muscle vepesitioning in 4. Mitral anuuloplasty was performed in all cases. Before and after surgery, cardiac function parameters were compared. Results No operative deaths occurred. One case wastransfered to mitral valve replacement due to regurgation, lntraoperative transesophageal echocardiography showed no mitral regurgitation in 38 and small regurgitation in 6 cases. The mean mitral valve effective orifice area(MVEOA) was (2.8±0.6) cm~2, with a mean gradient pressure of (6.21±1.34) mm Hg after operation. The mean followed up was ( 18.0±2.1 ) months. Echocardiography study showed that no mitral regurgitation in 35 cases, slight regurgi-tation in 9, mean mitral effective orifice area was (2.5±0. 8 ) cm~2, mean gradient pressure of (7.21±0. 45 ) mm Hg, no one need reoperation. Postoperative cardiac functions were significantly improved: the average left ventricular end-diastolic diameter (LVEDD) was (48±7) mm [ preoperative (56±6) nun, P <0.05], ejection fraction (EF) was 0.51～0.24( preoperative 0.45± 0.23, P < 0.05 ), the average left atrium diameter ( LA ) was ( 50±11 ) mm [ preoperative ( 62±23 ) mm, P <0. 05 ]. The function of mitral valves was well performed. Conclusion Leaflet enlargement with autologous pericardium com-bined with mitral annuloplasty was effective in repairing of diseased mitral valve. The advantages of the procedure including simplicity, good compatibility, avoiding foreign body and no need for anticoagulation.

Objective To investigate the protectve effects and underlying mechanisms of deferroxamine on glutamate-induced injury in cultured hippocampal neurons.Methods Primarily cultured hippocampal neurons from fetal rat were used in a model of glutamate induced neurotoxicity.There were two experimental groups.Neurons were pretreated with deferroxamine before glutamate in the deferroxamine group, and neurons were treated with glutamate only in the control group.The morphological change was examined under microscope.Hoechst 33342 DNA staining method was used to study the ratio of condensed nuclei.The levels of lactate dehydrogenase (LDH), malonaldehyde (MDA) and hydroxyl radical were determined using biochemistry.The change in calcium signal was detected using microfluorescent technique.Results The neurons pretreated by deferroxamine had intact morphology with the ratio of condensed nuclei at 14% ± 6% compared to 58% ± 6% (t= 8.98, P ＜0.01 ) in the control group.LDH level was (36.42 ± 8.99) U/L in the deferroxamine group and was (68.06 ± 11.26) U/L in the control group ( t =3.25,P＜0.05).The respective levels of hydroxyl radical were (34.21 ±4.23) U/L and (47.06 ±8.79) U/L (t = 3.11, P ＜0.05 ).The respective levels of MDA were (12.26 ± 2.78 ) nmol/mg and (28.86±5.19) nmol/mg(t =4.88,P＜0.01).Conclusion Deferroxamine can protect neurons from glutamate induced damage.The mechanisms include an inhibition of Ca2+ overload and reduction in the levels of MDA and hydroxyl radicals.

Objective To investigate the impact of polyclonal neural cell adhesion molecule antibody (P-NCAM-Ab) on the potency of botulinum toxin A (BTX-A).Methods Ninety male Sprague-Dawley rats were randomly divided into 3 equal groups:a normal control group,a BTX-A group and a P-NCAM-Ab group.The rats in the normal control group were injected with 100 μl of saline solution in their right gastrocnemius,while those in the BTX-A and P-NCAM-Ab groups were injected with 100 μl of BTX-A (0.5 U).In addition,the rats in the P-NCAM-Ab group were also injected with 100 μl of P-NCAM-Ab (the dosage was 20 U) at the same site on the 3rd day after the BTX-A injection.The rats' gastrocnemius muscle strength was evaluated with a self-made system for evaluating neuromuscular function before and after the toxin injection,on the 3rd day,as well as 1,2,4,6,8,10 and 12 weeks after the BTX-A injection.Any wet weight changes in the muscles were observed,and immunochemistry methods were employed to observe any structural changes in the motor endplates and nerve fibers at the different time points.Results After the saline injection,the average gastrocnemius muscle strength of the control group increased with time,while strength in the BTX-A and P-NCAM-Ab groups demonstrated a decrease in strength followed by a gradual increase.The average gastrocnemius muscle strength of the rats in the BTX-A and P-NCAM-Ab groups was significantly lower than that of the control group at all time points.Compared with the BTX-A group,the muscle strength of the P-NCAM-Ab group rats decreased further.Strength recovery in the BTX-A and P-NCAM-Ab groups was significantly slower than in the control group.The wet weight percentage in the BTX-A and P-NCAM-Ab groups at first decreased and then recovered with time.After the BTX-A injection,the average wet weight percentage of the P-NCAM-Ab group rats was significantly lower than that of the BTX-A group after 3 days,and 1,2 and 4 weeks.Karnovsky-Roots AchE staining showed that the motor endplates' color in the BTX-A and P-NCAM-Ab groups deepened gradually,though the color of the P-NCAM-Ab group was lighter than that of the BTX-A group at each time point.The mean optical density of the motor endplates' positive reaction area increased with time in both groups,but the P-NCAM-Ab group was lower than that of the BTX-A group at 1,2,4,8 and 12 weeks.Counting the nerve fibers dyed by gold chloride showed similar trends with both experimental groups significantly different from the control group.Conclusion P-NCAM-Ab can increase the potency of BTX-A and prolong its action.

Objective To develop a method for dynamically observing the biological efficacy of botulinum toxin A (BTX-A) and to investigate the dose-effect relationship between BTX-A dosage and muscle strength.MethodsFifty-four male Sprague Dawley rats were randomly divided into 9 groups.Groups 1-7 were injected intramuscularly with 0.1 ml BTX-A (0.01 U to 4.0 U) into the gastrocnemius on the right side.Rats in group 8 were injected intramuscularly with an equal volume of saline solution as the control group,and group 9 was used to determine the location of injection.Gastrocnemius muscle strength was evaluated using a self-made evaluation system before and after the toxin injection and on the 3rd,7th,14th,21st,30th,45th,60th and 75th day following.ResultsMuscle strength reached its lowest level on days 3 to 7,with a significant difference in the decline of muscle strength between the test groups and the control group up to day 60.With the lower BTX-A doses (0.01 U,0.1 U,0.5 U,1.0 U),muscle strength had decreased significantly on the 21st day,but recovered to its initial levels in all groups at the same time.There was no significant difference among the 1.0 U,1.5 U,2.0 U and 4.0 U groups.ConclusionsStandardized gastrocnemius injection combined with neuromuscular functional evaluation can establish a model of BTX-A dosage and muscle paralysis which can be used to assess the evolution of the biological efficacy of BTX-A.

The purpose of the study was to investigate the impact of rat cytomegalovirus(RCMV) infection on the development of the nervous system in rat embryos,and to evaluate the involvement of Wnt signaling pathway key molecules and the downstream gene neurogenin 1(Ngn1) In RCMV infected neural stem cells(NSCs).Infection and control groups were established,each containing 20 pregnant Wistar rats.Rats in the infection group were inoculated with RCMV by intraperitoneal injection on the first day of pregnancy.Rat E20 embryos were taken to evaluate the teratogenic rate.NSCs were isolated from E13 embryos,and maintained in vitro.We found:1) Poor fetal development was found in the infection group with low survival and high malformation rates.2) The proliferation and differentiation of NSCs were affected.In the infection group,NSCs proliferated more slowly and had a lower neurosphere formation rate than the control.The differentiation ratio from NSCs to neurons and glial cells was significantly different from that of the control,showed by immunofluorescence staining.3) Ngn1 mRNA expression and the nuclear β-catenin protein level were significantly lower than the control on day 2 when NSCs differentiated.4) The Morris water maze test was performed on 4-week pups,and the infected rats were found worse in learning and memory ability.In a summary,RCMV infection caused abnormalities in the rat embryonic nervous system,significantly inhibited NSC proliferation and differentiation,and inhibited the expression of key molecules in the Wnt/β-catenin signaling pathway so as to affect NSCs differentiation.This may be an important mechanism by which RCMV causes embryonic nervous system abnormalities.

Objective:To investigate the clinical efficacy of 125I seed implantation combined with anlotinib hydrochloride in the treatment of non-small cell lung cancer (NSCLC). Methods:61 cases of NSCLC patients were enrolled, of which 30 cases (observation group) received 125I seed implantation combined with anlotinib treatment, and 31 cases (control group) received 125I seed implantation only. To evaluate the curative effect and adverse reactions of all patients, the carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA21-1), neuroendocrine enolase (NSE), squamous cell carcinoma antigen (SCC) in the peripheral blood of the two groups was measured before the treatment and at 1 and 3 months after the treatment. Results:The effective rates in the observation group were 90.00% and 93.33%, the effective rates in the control group were 67.74% and 74.19% at 1 and 3 months after the treatment, respectively, and the difference in efficacy between the two groups was statistically significant ( χ2=4.504, P=0.034 vs. χ2=4.075, P=0.044). There was no significant difference in the incidence of adverse reactions between the two groups of patients after treatment ( P=0.785). At 1 and 3 months after the treatment, the levels of CEA, CYFRA21-1, NSE and SCC in the peripheral blood of the two groups of patients were lower than those before the treatment (all P<0.05). Conclusions:125I seed implantation combined with anlotinib hydrochloride is safe for the treatment of advanced non-small cell lung cancer, and has promotion value.