Objective To explore epicanthus correction by transverse incision method and concurrent double-eyelid incision operation.Methods The transverse straight-line incision was performed in new inner canthus to the original canthal corner point; after the original inner canthus corner point was reached,the oblique-line parallel incision was performed along the lower eyelid so that full subcutaneous separation was obtained on the upper and lower incision; the malpositioned isomerous orbicular muscle and thickened tissue were released and excised so that the epicanthus skin was naturally restored; the superficial head of the inner canthus ligament was folded and sutured when necessary ; after small cat ears at the temporal side of the inner canthal incision was removed,routine double eyelid plasty was then performed,but the double eyelid and the inner canthus incisions were not continuous.Results The procedures were clinically applied in 258 cases and followed up for 3-18 months,showing that the epicanthus has disappeared,new inner canthus was in good appearance,the scar was not obvious,recurrence of the epicanthus was not found and double eyelids were beautiful in appearance.Conclusions The epicanthus correction by transverse incision and concurrent double-eyelid plasty is a simple and ideal approach for correction of single-edged eyelid with epicanthus and it is worthy of clinical promotion.

This study was aimed to isolate and characterize of chemical constituents in a traditional Mongolian medicine HERBA CLEMATIS. Normal and reverse phase coloumn chromatography, gel filtration chromatography sephadex LH-20, and preparative HPLC were used for isolation and purification compounds from the water extraction of the arial part ofC. aethusaefoliaTurcz. The planar structures and spatial configurations of isolated compounds were identified by high resolution MS, 1D and 2D NMR, and other spectrographic methods. The chemical research on the Mongolian medicine results 6 compounds, dihydrodehydrodiconiferyl alcohol (1), syringaresinol (2), pinoresinol (3), epi-pinoresinol (4), lirioresinol B dimethyl ether (5) and loliolide (6). All the compounds were isolated from this plant for the first time.

Aim To study the protective effect of CDPS on acute aging mouse model induced by D-galactose (D-gal) and its mechanism.Methods (1) The acute aging mouse model was induced by D-gal.After CDPS (25、50、100 mg·kg-1) treatment, the improving effect on learning and memory in mice was examined in vivo.(2) We also established the aging model on PC12 cells in vitro.After CDPS treatment (150、200 mg·L-1), the level of p-CREB in the nucleus was detected by Western blot, and the content of cAMP, PKA and brain derived neurotrophic factor (BDNF) levels were examined by the Elisa kits.Moreover, cAMP, PKA and BDNF were detected in PC12 cells under the condition that H89, the inhibitor of PKA, co-cultured with PC12 cells after CDPS treatment.(3) The UPLC/Q Exactive MS method was developed for determining the concentration of glutamic acid, dopamine and norepinephrine, which secreted in PC12 cells after CDPS treatment.Results (1) In vivo, CDPS significantly improved the memory impairment in aging mice induced by D-gal in the Morris assay.(2) In vitro, CDPS could significantly increase the expression of p-CREB (P<0.05), PKA, cAMP and BDNF (P<0.05).The H-89 abolished the increase of p-CREB (P<0.05), PKA, cAMP and BDNF (P<0.05) in PC12 aging cells induced by D-gal after CDPS treatment.(3) CDPS increased the release of dopamine, norepinephrine, and glutamate secreted in PC12 cells.Conclusion CDPS could significantly improve the learning and memory ability on aging mouse model in vivo, and reversed the damage in PC12 cells induced by D-gal by activating cAMP/PKA/CREB signal cascade, increase the expression of BDNF, and increasing modestly the release of excitatory neurotransmitter.

OBJECTIVETo investigate the effect of the vector carrying short hairpin RNA targeting epidermal growth factor receptor (shRNA-EGFR) on the radiosensitivity of human nasopharyngeal carcinoma xenografts in nude mice.

METHODSshRNA-EGFR was transfected into human nasopharyngeal carcinoma cell line CNE1 via Lipofectamine 2000. The transfected cells were collected for quantitative RT-PCR detection of the expression level of EGFR mRNA. Western blotting was used to examine the expression of EGFR protein. CNE1 cells were inoculated into nude mice and the tumor volume was measured every 2 days. shRNA-EGFR was intratumorally injected in the mice, and 16 days after radiotherapy, the mice were sacrificed and tumors examined for radiosensitivity.

RESULTSshRNA-EGFR was effectively delivered via Lipofectamine 2000 into CNE cells to result in a significant downregulation of EGFR mRNA and protein expressions (P<0.05). A significant difference was noted in the tumor volume and weight in the tumor-bearing nude mice between shRNA-EGFR plus radiotherapy group and the control, exclusive radiotherapy and shRNA-EGFR groups (P<0.05).

CONCLUSIONshRNA-EGFR combined with radiotherapy can effectively inhibit the growth of nasopharyngeal carcinoma in nude mice. shRNA-EGFR can enhance sensitivity of nasopharyngeal carcinoma to radiotherapy.

Animals ; Gene Expression Regulation, Neoplastic ; Mice ; Mice, Nude ; Nasopharyngeal Neoplasms ; genetics ; radiotherapy ; RNA, Catalytic ; genetics ; RNA, Small Interfering ; genetics ; Radiation Tolerance ; genetics ; Receptor, Epidermal Growth Factor ; genetics ; Transfection ; Xenograft Model Antitumor Assays

A method for simultaneous determination of PCDDs, dl-PCBs, BFRs and PBDD/Fs in flue gas from stationary source was developed. The sample was extracted by Soxhlet apparatus with toluene, and followed by purification through sulfuric acid partition and multi-layer silica gel column separation. The target compounds were then all separated by passing through the active carbon-dispersed silica gel column and reversal eluting. Gas chromatography coupled with a thermostable capillary column ( short length, thin stationary phase film) was operated at pulse injection mode. High resolution mass spectrometry set at low-electron-energy ionization was used for quantification. The high- and low-brominated compounds were determined simultaneously. The detection limits of this method were 0. 081-1. 2 pg for PCDD/Fs, 0. 10-0. 32 pg for dl-PCBs, 0. 14-12 pg for PBDEs, 0. 26-16 pg for new BFRs, 0. 44-3. 6 pg for tetra- to hepta-BDD/Fs and 8. 2-12 pg for OBDD/F. Recoveries ( RSDs) in spiked flue gas samples were 88%-115%(2. 9%-6. 1%) for PCDD/Fs, 84%-118% (3. 2%-10%) for dl-PCBs, 71%-135% (2. 1%-18%) for PBDEs, 71%-114% (2. 9%-7. 4%) for new BFRs, 83%-127% (5. 2%-10%) for tetra-to hepta-BDD/Fs and 52%-149% ( 23%-24%) for OBDD/F. All quality control data fell within the acceptable range specified in analysis standards for flue gas.

Objective To explore the effect of erythropoietin (EPO) attenuating apoptosis in old rat hippocampal neuronal cells exposed to sevoflurane and the role of toll like receptor 4.Methods Twenty months old SD rats,male,550-750 g,in accordance with the random number table,were divided into 3 groups (n =9):control group (group C),sevoflurane treatment (group S),and sevoflurane plus EPO treatment (group ES).The rats in group S and ES were subjected to inhale 4％ sevoflurane for 6 h,but the rats in group C were inhaled air-oxygen only.The rats in group ES were injected with EPO into caudal vein at 24 h,48 h,and 72 h after sevoflurane exposure.The cognitive ability was assessed by Morris water maze test;the effects of hippocampal apoptosis were assessed by TUNEL assays;the expressions of TLR4 mRNA was measured by RT-PCR assay;mitochondrial membrane potential (MMP) was assessed by JC-1 fluorescence;the expressions of APP and Aβ were assessed by western blot.Results Compared with group C,there were significant increases of escape latency period,neuronal apoptosis,TLR4 mRNA,and APP and Aβ expression,but a decrease of MMP in group S (P＜0.05).Compared with group S,there were significant decreases of escape latency period,neuronal apoptosis,TLR4 mRNA,and APP and Aβ expression,but a increase of MMP in group ES (P＜0.05).Conclusion The attenuation of rat hippocampal neuronal apoptosis induced by EPO could be associated with inhibition of TLR4,improvement of MMP,as well as inhibition of APP and Aβ activity.

OBJECTIVE:To establish HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry for rapid determination of aconitine,mesaconitine,hypaconitine,benzoylaconitine,benzoylmesaconine and benzoylhypacoitine in rat plasma. METHODS:Internal standard lappaconitine was added into plasma sample,and methanol precipitated protein was used for pretreatment. HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry was adopted. HPLC condition was as follows as Sinochrom ODS-BP C18column,mobile phase consisted of acetonitrile-1% formic acid solution(50:50,V/V),the flow rate of 0.6 mL/min,sample size of 10 μL,column temperature of 25 ℃,automatic sampler temperature of 4 ℃. Mass spectrum scanning mode was full ion monitoring model,positive ion acquisition,mass charge ratios(m/z)of ion were 646.32(aconitine), 632.30(mesaconitine),616.31(hypaconitine),604.31(benzoylaconitine),590.29(benzoylmesaconine),574.30(benzoylhypacoitine), 585.31(internal standard). Six male Wistar rats were collected and given single dose of total alkaloid extract of Aconitum carmichaeli(4 mg/kg)intragastrically. Blood samples were collected before medication(0 h)and 0.5,0.75,1.25,1.5,2,4,6, 8,10,24 h after medication. Plasma concentration was determined and pharmacokinetic parameters were calculated by using PK-Solver V2.0 software. RESULTS:The linear range of 6 kinds of aconitum alkaloids in plasma were 0.1-10 μ g/L(r>0.992). The limit of quantitation was 0.1 μ g/L. Average recovery was higher than 75%,RSDs of intra-day and inter-day,matrix effects,stability test were all lower than 15%. The tmaxof 6 kinds of aconite alkaloids were about 1.2 h;t1/2were about 10 h;cmaxof monoestertype aconite alkaloids were higher than those of diester-type aconite alkaloids. CONCLUSIONS:Established HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry is accurate,sensitive,simple and rapid, and can be used for plasma concentration monitoring of 6 kinds of aconitum alkaloids.

Objective To compare the application values and setup errors between vacuum bag plus body mask and customized alpha cradle duringproton and carbon therapy using Siemens 6D robotic couch in prostate cancer patients.Methods Nineteen patients received vacuum bag plus body mask setup were allocated into the vacuum bag group andl9 patients with alpha cradle were assigned into the alpha cradle group.Orthogonal X-ray portals were performed to verify the treatment position before beam delivery in every fraction.The couch correction between the portal and reference DRR through manual image registration was recorded as setup errors in 6 directions including the lateral,supine-inferior,anterior-posterior,yaws,roll and pitch,respectively.Two-tail t-test was used to analyze the setup error data from each direction between two groups.Results In total,452 and 436 sets of data errors were collected from the vacuum bag and alpha cradle groups.The average setup errors and standard deviation in the vacuum bag and alpha cradle groups in the lateral,supine-inferior,anterior-posterior,yaws,roll and pitch directions were (0.63±0.48) cm vs.(0.33±0.24) cm (P=0.000),(0.40±0.3) cm vs.(0.31±0.25) cm (P=0.000),(0.69±0.61) cm vs.(0.82±0.69) cm (P=0.006),0.65°±0.47°vs 0.32°±0.25°(P=0.000),1.05°±0.95°vs 1.16°±0.94° (P=0.100) and 0.67°±0.56°vs 0.40°±0.36° (P=0.000),respectively.The maximum setup errors were detected in the pitch direction for both groups.Conclusions During the proton and carbon therapy using Siemens 6D robotic couch,two setup methods using vacuum bag plus body mask and customized alpha cradle should be selected according to the individual conditions of patients.A customized foot fixer should be utilized to reduce the uncertainty in the femoral head region.

The effects of testosterone on norepinephrine release were investigated in the isolated rat hearts.Sprague-Dawley male rats (n=120) were randomized to testosterone and control groups.The rats in testosterone group were perfused with modified Krebs-Henseleit buffer containing different concentrations of testosterone (0.1,1.0,10.0,and 100.0 nmol/L,respectively).Myocardial ischemia was induced by globally stopping the perfusion flow.Exocytotic norepinephrine release was induced by electrical field stimulation at 5 V (effective voltage) and 6 Hz (pulse width of 2 ms) for 1 min.The overflow of norepinephrine was determined by high pressure liquid chromatography and electrochemical detection (HPLC-EC).Following acute ischemia,testosterone (1.0,10.0 and 100.0 nmol/L) significantly reduced norepinephrine release (P＜0.01),and the norepinepherine overflow was similar between the control and 0.1 nmol/L testosterone group (P＞0.05).Electrical stimulation of the ventricle evoked norepinepherine release,and this was diminished by the perfusion with testosterone at the concentrations of 1.0,10.0 and 100.0 nmol/L (P＜0.01).It is suggested that testosterone suppresses ischemia- and electrical stimulation-induced norepinepherine release in the isolated rat hearts.

Objective To evaluate the role of mitochondrial ATP sensitive potassium ( mito-KATP ) channel in dexmedetomidine-induced inhibition of subarachnoid hemorrhage ( SAH )-caused programmed cell death ( PCD) in cardiomyocytes of rats. Methods On hundred and twenty clean-grade healthy male Sprague-Dawley rats, aged 9-10 weeks, weighing 350-400 g, were divided into 5 groups ( n=24 each) using a random number table method: sham operation group ( group Sham ) , SAH group, SAH plus dexmedetomidine group ( group SD) , 5-HD plus SAH and dexmedetomidine group ( group HSD) and 5-HD plus SAH group ( group HS) . The rats were subjected to SAH by intracranial vascular puncture after being anesthetized with pentobarbital sodium. Dexmedetomidine 5 μg∕kg was infused for 10 min via the jugular vein starting from the time point after intracranial vascular puncture, followed by a continuous infusion of 5μg·kg-1 ·h-1 for 1 h in SD and HSD groups. 5-HD 30 mg∕kg was intraperitoneally injected at 1 h before intracranial vascular puncture in HSD and HS groups. Blood samples were collected from the abdominal aor-ta at 24 h after intracranial vascular puncture for determination of serum cardiac troponin I ( cTnI) concen-trations. The animals were then sacrificed, and myocardial specimens were collected for determination of PCD rate ( by TUNEL) , reactive oxygen species ( ROS) activity ( by DCFH-DA assay) , and expression of cleaved caspase-3, cleaved caspase-1 and interleukin-1beta ( IL-1β) ( by Western blot) . Results Com-pared with group Sham, the serum concentrations of cTnI, PCD rate and ROS activity were significantly in-creased, and the expression of cleaved caspase-3, cleaved caspase-1 and IL-1βwas up-regulated in SAH, SD, HSD and HS groups ( P<0. 05) . Compared with group SAH, the serum concentrations of cTnI, PCD rate and ROS activity were significantly decreased, and the expression of cleaved caspase-3, cleaved caspase-1 and IL-1βwas down-regulated in group SD, and the serum concentrations of cTnI, PCD rate and ROS activity were significantly increased, and the expression of cleaved caspase-3, cleaved caspase-1 and IL-1βwas up-regulated in group HS ( P<0. 05) . Compared with group SD, the serum concentrations of cT-nI, PCD rate and ROS activity were significantly increased, and the expression of cleaved caspase-3, cleaved caspase-1 and IL-1β was up-regulated in group HSD ( P<0. 05) . Compared with group HSD, the serum concentrations of cTnI, PCD rate and ROS activity were significantly increased, and the expression of cleaved caspase-3, cleaved caspase-1 and IL-1βwas up-regulated in group HS ( P<0. 05) . Conclusion mito-KATP channel is involved in dexmedetomidine-induced inhibition of PCD in cardiomyocytes of rats with SAH.