Objective To observe the effect of epidural injection with different dose of butorphanol on the rats' neurological function.Methods A PE-530 catheter was inserted into the epidural space of all the SpragueDawley rats (male, weighting 180 ～210 g) at L1-2 level.After three days, a total of 32 rats without any motor dysfunction were randomly divided into 4 groups as follows saline(NS) group (group C, n= 8 )and butorphanol injection (B) group( B1∶ n=8;B2∶ n=8;B3∶ n=8).Rats in group C were epidurally injected NS 30 μl each ,and rats in group B1, B2 and B3 were respectively epidurally injected Butorphanol 60 μg/30μl, 120 μg/30 μl,240 μg/30 μl (all diluted with NS) ,and 1 time per day for5 days.The neurological function of rats was recorded before injection (T0) and 6h after injection on day 1 ～4(T1 ～T 4) and 6h,24h and 72h after injection on day 5 (T5 ～T7) by BBB (BASSO,BEATTIE and BRESNAHAN ) Score and the inclined plane test .Results Compared with group C ,the BBB score and the inclined plane test of group B1 showed no significant difference throughout the experimental period(P＞ 0.05 ).There was also no significant difference at T0 ～ T3 of group B2 and group B3 compared with group C (P ＞ 0.05 ), while at T4, the BBB score ( ( 18.50 ± 2.00 ) points, ( 16.38 ± 2.33 ) points) and the inclined plane test( (58.75 ± 5.17 )°, (59.38 ± 3.20) ° ) of the two groups were both obviously decreased when compared with group C( (21.00 ±0.00) points, (65.00 ±3.78)°, P＜0.05) ,and the same significant differences appeared at T5,T6 and T7 (P ＜ 0.05 ).Conclusion Repeated epidural injection of butorphanol 60 μg have no effect on neurological function of rats,while repeated epidural injection of butorphanol 120 μg and 240 μg could impaire the neurological function.

Ten glucosyloxybenzyl 2-isobutylmalates and one benzyl alcohol glycoside were isolated from the dry tuber of Pleione bulbocodioides, which is a specie of Orchidaceae family and its dry tuber is one of the main sources of traditional Chinese medicine "shanci-gu", by a combination of various column chromatographic methods, including ODS, macroporous adsorbent resin, Sepheadex LH-20, and preparative HPLC. Their structures were identified on the basis of chemical evidences and spectroscopic analysis asloroglossin (1), grammatophylloside A (2), cronupapine (3), (-)-(2R, 3S)-1-(4-β-D-glucopyranosyloxybenzyl)-4-methyl-2-isobutyltartrate (4), vandateroside II (5), grammatophylloside B (6), bis [4-(β-D-glucopyranosyloxy) -benzyl] (S) -2-isopropylmalate (7), gymnoside I (8), militarine (9), dactylorhin A (10), gastrodin (11). Compounds 1-7 were isolated from this genus for the firt time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Malates ; chemistry ; isolation & purification ; Molecular Structure ; Orchidaceae ; chemistry ; Spectrometry, Mass, Electrospray Ionization

This study was to investigate the chemical constituents from pseudobulbs of Pleione yunnanensis, one of the source of traditional Chinese medicine "Shancigu". The chemical constituents were isolated by various chromatography methods, including silica gel, ODS, Sephadex LH-20, and semi-preparative HPLC. Fourteen compounds were isolated and identified from the EtOAc fraction of 90% ethanol extract, including five dihydrophenanthrenes, four bibenzyls, two triterpenoids, and three phenylacrylic acids. Their structures were identified on the basis of the spectral data as 4, 7-dihydroxy-2-methoxy-9,10-dihydrophenanthrene (1), 4, 7-dihydroxy-1-(p-hydroxybenzyl)-2-methoxy-9,10-dihydrophenanthrene (2), (2,3-trans)-2-(4-hydroxy-3-methoxyphenyl) -3-hydroxymethyl-10-methoxy-2,3,4,5-tetrahydro-phenanthro[2,1-b]furan-7-ol (3), pleionesin B (4), blestriarene A (5), batatasin III (6), 3, 3'-dihydroxy-2-(p-hydroxybenzyl) -5-methoxybibenzyl (7), 3', 5-dihydroxy-2-(p-hydroxybenzyl) -3-methoxybibenzyl (8), 3,3'-dihydroxy-2,6-bis(4-hydroxybenzyl) -5-methoxybibenzyl (9), triphyllol (10), pholidotin (11), (E) -p-hydroxycinnamic acid (12), (E)-ferulic acid (13), and (E)-ferulic acid hexacosyl ester (14). Compounds 5,10-14 were separated from this plant for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Orchidaceae ; chemistry ; Spectrometry, Mass, Electrospray Ionization

To establish an HPLC method for determination of dactylorhin A and militarine in Cremastrae Pseudobulbus/Pleiones Pseudobulbus. The analysis was achieved on an Alltech Prevail C18 column (4. 6 mm x 250 mm, 5 microm) using a mobile phase of acetonitrile (A), water (B) gradient elution in a total run time of 35 min (0 min, 20:80; 30 min, 55:45; 35 min, 55:45) and a diode array detector was set at 224 nm. The flow rate was 0.8 mL x min(-1). The assay displayed good linearity over the concentration range of 0.257-9.95 microg (r = 0.999 8), and 0.128-10.27 microg (r = 0.999 9), respectively. The average recoveries (n = 9) were 94.70% and 102.8% for dactylorhin A and militarine, respectively. The method is accurate, quick, simple and reproducibility. It can be used for the quality control of Pleione bulbocodioides and Pleione yunnanensis.
Chromatography, High Pressure Liquid ; Glucosides ; analysis ; Orchidaceae ; chemistry ; Succinates ; analysis

Fourteen compoumds were isolated from the ethyl acetate portion of the 95% ethanolic extract of Pleione bulbocodioides by a combination of various chromatographic techniques including silica gel, ODS, macroporous adsorbent resin, Sephadex LH-20, and preparative HPLC, of which ten compoumds were phenanthrenes and dihydrophenanthrenes, two compoumds were bibenzyls, one was lignan and a sterol. Their structures were identified on the basis of spectroscopic data as monbarbatain A(1), 2, 7, 2'-trihy-droxy-4, 4', 7'-trimethoxy-1, 1'- biphenanthrene(2), blestriarene A(3), pleionesin B(4), shanciol H(5), 17-hydroxy-7'-(4'-hy-droxy-3 '-methoxyphenyl)- 4-methoxy-9, 10, 7', 8'-tetrahydrophenanthro[2, 3-b]furan-8'-yl methyl acetate(6), 1-p-hydroxybenzyl-4-methoxy phenanthrene-2, 7-diol(7), 1-p-hydroxybenzyl-4-met-hoxy-9, 10-dihydrophenanthrene-2, 7-diol(8), hircinol(9), coelonin( 10), gigantol(11), batatasin 11 (12), syringaresinol(13) and ergosta4, 6, 8 ( 14) , 22-tetraen-3-one (14). Compounds 1-3, 9, 13 and 14 were isolated from this genus for the first time.
Drugs, Chinese Herbal ; chemistry ; Orchidaceae ; chemistry ; Organic Chemicals ; analysis

ObjectiveTo investigate the effect of patient-controlled epidural analgesia(PCEA) with opioids on serous myelin basic protein(MBP) and somatosensory evoked potential(SEP) of lower limbs in puerperants.MethodsA total of 120 puerperants,after receiving cesarean section,were divided into four groups by random number table method as group B,BR,MR and R randomly,and each group included 30 cases.After surgery,each case received PCEA:group B received 0.008％ butrophanol;group BR received 0.008％ butrophanol + 0.2％ ropivacaine;group MR received 0.004％ morphine +0.2％ ropivacaine and group R received 0.2％ ropivacaine only.VAS score,OAA/S score,adverse effect occurrence,concentration changes of serous MBP,SEP of both lower limbs and neurological function were observed at 2h(T1 ),4h(T2),8 h(T3),12h(T4),24 h(T5) and 48h (T6) after surgery.ResultsVASscoresofgroupBR(1.64±0.38,1.86±0.62,1.93±0.67) and MR( 1.74 ±0.39,1.91±0.58,1.98 ±0.63) at T3,T4,T5 were lower than those of group B(4.6 ±0.5,4.6 ±0.3,4.7 ±0.3)and R(2.64 ±0.41,2.83 ±0.91,3.37 ±0.87) (P＜0.05).There was no significance in four groups in OAA/S score at each point (P ＞ 0.05 ).Incidence of nausea ( 6 cases),vomiting ( 2 cases) and abdominal distention ( 5cases) of group M was higher than that of other three groups(P＜0.05).There was no significant difference in concentrations of serous MBP,SEP and neurological function in all four groups between preoperative time and 48h after operation(P＞0.05).ConclusionLower-dose and lower- concentration opioids used for PCEA have no influence on serum MBP and SEP.

Objective: To construct the expression plasmid of a novel gene human NBEAL1 (neurobeachin like 1), and to study its relationship with the pathological grades of glioma. Methods: Total RNA of human glioma cell line U251 was extracted. NBEAL1 expression plasmid pGEX-KG/NBEAL1 was constructed and transferred into E. coli BL21. Recombinant NBEAL1 protein was induced by IPTG and further purified by GST affinity chromatographic column. The purity of recombinant NBEAL1 protein was examined by Western blotting analysis. A NBEAL1 protein specific monoclonal antibody was prepared and was used to study the relationship of NBEAL1 expression with pathological grades of glioma. Results: The NBEAL1 gene fragment was successfully cloned into pGEX-KG expression plasmid and verified by DNA sequencing. The recombinant NBEAL1 protein was expressed in inclusion bodies, with a yield of more than 30% of total bacterial proteins; the purity of purified NBEAL1 protein was above 95%. Western blotting analysis confirmed that the purified protein containing GST tag and NBEAL protein. NBEAL1 protein was lowly expressed in normal brain tissues and highly expressed in low grade glioma tissues; and the expression of NBEAL1 decreased with the increase of glioma malignancy. Conclusion: The NBEAL1 protein has been successfully cloned, expressed and purified. NBEAL1 protein expression in glioma tissues is negatively associated with the pathological grades of glioma.

Objective:To investigate the inhibitory effect of breast cancer-associated antigen 1(BRCAA1)gene silencing on gastric cancer MGC-803 cells and the related mechanism.Methods:Plasmid shRNA-BRCAA1 and shRNA-N were constructed and transfected with FuGene HD into gastric cancer cell line MGC-803.The transfection efficiency was examined using fluorescent microscope 24 h later.The total RNAs was extracted 48 h 'after transfection and the expression of BRCAA1 and GAPDH gene were analyzed by real-time PCR.The cell proliferation was assessed by MTT assay 24 h,48 h,and 72 h after transfection.The cell apoptosis was determined by Annexin V-PE/TAAD.The expression of Rb,Bax, Bcl-2 and BRCAA1 proteins was analyzed by Western blotting 48 h after transfection.Results:We found that the transfection efficiency of shRNA-BRCAA1 was(81.2?2.6)%24 h after transfection.Forty-eight hours after transfection with shRNA-BRCAA1 the expression of BRCAAI mRNA decreased by 61.4%;the inhibition rate of MGC-803 cells growth was 45.0%.The cell apoptosis rate of shRNA-BRCAA1 transfection group was significantly higher than those of untransfected group and mock plasmid transfected group([14.4?1.6]%vs[5.4?2.0]%,[4.4?2.5]%,P

Objective:To investigate the clinical features of direct and indirect carotid cavernous fistulas (CCFs).Methods:Patients with CCF treated in the Affiliated Hospital of Xuzhou Medical University from January 2010 to August 2020 were enrolled retrospectively. Relevant clinical data were collected, including the main clinical manifestations, neuroimaging features, and treatment methods. The clinical features of direct and indirect CCFs were compared.Results:A total of 31 patients were enrolled in the study, 29 (93.5%) had ocular symptoms, of which conjunctival hyperemia and edema ( n=24, 77.4%), exophthalmos ( n=19, 61.3%) and orbital murmur ( n=18, 58.1%) were most common. There were 23 patients (74.2%) in direct CCF group and 8 (25.8%) in indirect CCF group. The former had more history of head trauma (78.2% vs. 12.5%; P=0.002), more flow volume (high-flow CCFs: 100% vs. 37.5%; P<0.001) and more likely to cause orbital murmur (69.6% vs. 25.0%; P=0.043). Endovascular embolization was safe and effective. The common methods of endovascular embolization were EVAL glue combined with coil embolization ( n=18, 66.7%) and detachable balloon embolization alone ( n=6, 22.2%). Conclusion:Ocular manifestations are most prominent in patients with CCFs. Direct CCF is more common, usually with a history of head trauma, and the clinical and imaging features are more typical. Interventional embolization is the preferred treatment option for patients with CCF.

Objective: To prepare a nanoprobe, anti-human melanoma ganglioside single chain variable fragment (GD/ScFvMEL) antibody conjugated with CdTe quantum dot, and to observe its ability to specifically bind human malignant melanoma cells. Methods: The GD/ScFvMEL gene was cloned into pET32a (+), and the plasmid was then transformed into E. coli BL21 (DE3) for GD/ScFvMEL protein antibody expression. The expressed GD/ScFvMEL antibody was purified by denaturing method and further refolded by modified dialysis method. The purified GD/ScFvMEL antibody was analyzed by SDS-PAGE. The GD/ScFvMEL-QDs nanoprobe was prepared by conjugating GD/ScFvMEL antibody with CdTe quantum dot, and its specificity was observed by incubating with MGC-803 cells and melanoma A375 cells. Results: The recombinant pET32a-GD/ScFvMEL was constructed and confirmed by PCR, restriction endonuclease analysis and DNA sequencing. The proportion of expressed GD/ScFvMEL antibody in total bacteria proteins was about 40% as detected by SDS-PAGE. The purified- and refolded-GD/ScFvMEL antibody was effectively conjugated with CdTe quantum dot, and the resulting GD/ScFvMEL-QDs nanoprobe was successfully prepared. The GD/ScFvMEL-QDs nanoprobe could specifically bind melanoma A375 cells, but could not bind stomach cancer MGC-803 cells. Conclusion: We have successfully prepared an anti-human melanoma ganglioside single-chain antibody-CdTe quantum dot nanoprobe, which can specifically bind melanoma cells.