By functional plates,16 strains which can produce?-mannana-se were isolated frnm 28 Bacillus spp.Using a pair of degenerated primers,the conserved fragments of?-mannanase gene from the selected strains were amplified by PCR.The obtained nucleotide fragments were sequenced and compared with the homologous?-mannanase genes in GenBank and a phylogenetic tree was generated.Comparing to the genes coding?-mannanase published,the cloned nucleotide fragments show the highest sequence identity between 62% and 98%.The genes coding fnr?-mannanase of Bacillus circulus have low identity while the?-mannanase genes of Bacillus subtilis and other Bacillus spp. have high identity.

Day-old chick is unique animal model in brain development and behavior study. The intermediate medial mesopallium (IMM), a region of the chick forebrain, is intimately involved in the early learning processes, which offers the ideal opportunity to study the neural changes that underlie behavioral plasticity. In this paper, the intracellular recordings were conducted from IMM neurons in chick forebrain slices, in which electrophysiological properties, synaptic responses and long-term potentiation (LTP) were observed. Coronal sections of left forebrains (500 mum thick), containing IMM, were prepared from domestic chicks, aged 2-10 days. In 69 IMM neurons, the resting membrane potential was measured to be (-59.4+/-5.3) mV, slope membrane resistance (70.8+/-27.2) MΩ, and time constant (10.2+/-4.3) ms. The amplitude, threshold, overshoot, half-width, max rise slope and max decay slope of action potential evoked by intracellular current injection were (85.2+/-9.4) mV, (-38.7+/-7.6) mV, (25.6+/-8.9) mV, (2.1+/-0.5) ms, (150.5+/-41.2) mV/ms and (-64.3+/-14.0) mV/ms, respectively. Spike-firing frequency was increased with depolarizing current intensity in 32 of 69 tested cells [linear regression slope was (21.5+/-10.9) Hz/nA, P<0.05 in all cells]. The depolarizing synaptic responses (i.e. EPSPs), with stimulus intensity- and membrane potential-dependent properties, were elicited by dorsal (n=25) or ventral (n=62) focal electrical stimuli at 0.1 Hz in all tested IMM neurons and could be nullified reversibly by perfusion with 100 mumol/L AP5 (NMDA receptor antagonist) and 3 mumol/L DNQX (non-NMDA receptor antagonist), but enlarged by 6 mumol/L bicuculline (GABA(A) receptor antagonist). The EPSPs evoked by ventral stimulation were persistently increased after tetanic stimulation (5 Hz, 300 pulses/train, 2 trains, train interval 10 min) in 6 of 12 tested IMM neurons. The amplitude of EPSPs was potentiated to more than 120% of control level (when analyzed at 45 min of enhancement, P<0.05, n=5), which lasted at least 30 min and then could be referred to as LTP. Moreover, area under curve, duration and max rise slope of EPSPs were also enhanced (P<0.05), while no significant changes were observed in the electrophysiological parameters of IMM neurons following induction of LTP (P>0.05). These results suggest that the intracellular recording techniques in the chick brain slices can be used to perform multi-parameter analysis of synaptic responses and their LTP.
Animals ; Brain ; physiology ; Chickens ; In Vitro Techniques ; Long-Term Potentiation ; Membrane Potentials ; Receptors, N-Methyl-D-Aspartate ; antagonists & inhibitors

Objective To explore the effect of spinal 360°circumferential resection decompression combined with bone cement fill-separation and pedicle screw fixation instrumentationb for patients suffering from spinal metastases.Methods Retrospectively analyzed the clinical data of 42 patients suffering from spinal metastases and underwent spinal 360°circumferential resection decompression combine with bone cement fill-separation and pedicle screw fixation instrumentation in our hospital from April 2012 to October 2016.The pain level were assessed by visual analogue scale(VAS),the functional impairment was classified by Karnofsky score(KPS) and the neurologic deficit was evaluated by Frankel grade before preoperation,1 week after operation and 3 months after operation.Results Through the following-up of 3 months,there was no neural symptoms relapse in primary segment.The VAS score was (0.57 ±0.79) points,and the KPS was (72.61 ± 19.12) points 3 months after operation,which were significantly improved compared with the data before operation (P ＜ 0.05).The VAS score improvement rate was (91.09％ ±13.73％),and the Frankel grade improvement rate was 80％.Conclusion Circumferential resection decompression combined with bone cement fill-separation and pedicle screw fixation instrumentationb for patients suffering from spinal metastases can effectively improve the neural symptoms and life-quality of patients and prevent neural symptoms relapsing in primary segment after operation.

Objective To explore the effect of action observation therapy (AOT) on upper limb function in children with spastic hemiplegic cerebral palsy,and the better program. Methods From March to November,2017,45 children with spastic hemiplegic cerebral palsy were randomly assigned to control group and AOT group.The control group was further divided into group A(n=11)and group B(n=11), and received conventional rehabilitation treatment and scenic-observation therapy, 20 minutes every time for group A,and 30 minutes every time for group B.AOT group was further divided into group C(n=10)and group D (n=11), and received AOT in addition, 20 minutes every time for group C, and 30 minutes every time for group D,five times each week for twelve weeks for all the groups.Before treatment,eight and twelve weeks af-ter treatment, they were assessed with hand grip strength, Upper Extremities Functions Test (UEFT), and Wee Functional Independence Measure(WeeFIM). Results After treatment,the hand grip strength and the score of UEFT were better in group D than in groups B and C(P<0.05),and were better at twelve weeks than at eight weeks(P<0.05).No significant difference was found in the score of WeeFIM among groups after treatment(P>0.05). Conclusion AOT could improve upper limp function in children with spastic hemiplegic cerebral palsy,and it's more ef-fective after more training.

OBJECTIVETo investigate the effect of Compound Qingqin Liquid (CQL) on the expression level of angiotensin II (Ang II) and COX-2 mRNA transcription and protein expression in the renal tissue of rats with uric acid nephropathy.

METHODSSD rats were randomly divided into the blank control group, the model group, the positive drug group, the high, moderate, and low dose CQL group according to number randomization principle. The model was established by gastrogavage of adenine, accompanied with yeast feeding. Distilled water was given by gastrogavage to rats in the blank control group and the model group. Allopurinol at the daily dose of 9.33 mg/kg was given by gastrogavage to rats of the positive control group. CQL at the daily dose of 3.77 g/kg, 1.89 g/kg, and 0.09 g/kg was respectively given by gastrogavage to rats in the high, moderate, and low dose CQL groups. All treatment lasted for 6 weeks. Rats were randomly divided at week 4 (3 in the blank control group, and 6 in the rest groups), and the rest rats were killed at week 6. The renal tissue was extracted. The expression level of Ang II and COX-2 mRNA transcription were detected by RT-PCR. The expression level of Ang II was detected by ELISA. The expression level of COX-2 protein was detected by Western blot and immunohistochemical assay.

RESULTSCompared with the blank control group, except the mRNA expression of Ang II at week 4, the mRNA and protein expression of Ang II and COX-2 obviously increased at week 4 and 6 in the model group (P < 0.01, P < 0.05). The COX-2 protein expression at week 4 was obviously lower in the high and moderate dose CQL groups than in the model group and the low dose CQL group (P < 0.05); the average integral of optical density value was obviously lower in the positive control group than in the model group. Except the mRNA expression of Ang II in the high dose CQL group at week 6, the mRNA and protein expression of Ang II obviously decreased in the positive control group and each dose CQL group (P < 0.01, P < 0.05). Of them, the effects were better in the high and moderate dose CQL groups than in the positive control group and the low dose CQL group (P < 0.05, P < 0.01). Besides, the mRNA expression of COX-2, the average integral of optical density value were obviously lower in the positive control group and each dose CQL group than in the model group (P < 0.05). The protein expression of COX-2 was obviously lower in the high and moderate dose CQL groups than in the model group (P < 0.05). Of them, the mRNA expression of COX-2 was better in the moderate dose CQL group than in the positive control group (P < 0.05); the protein expression of COX-2 was better in the high dose CQL group than in the low dose CQL group (P < 0.05).

CONCLUSIONCQL was capable of lowering the expression level of Ang II, COX-2 mRNA transcription and protein expression, thus suppressing the inflammatory pathological injury of the renal tissue.

Angiotensin II ; metabolism ; Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; metabolism ; Kidney Diseases ; drug therapy ; metabolism ; Male ; RNA, Messenger ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Uric Acid

OBJECTIVETo investigate the effect of compound qingqin liquid (CQL) on Toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in rats with urate nephropathy, and to explore its renal protection mechanism.

METHODSTotally 55 SD rats were randomly divided into 5 groups, i.e., the normal control group (n =5), the model group (n =10), the positive drug group (n=10), and the high-, medium-, low-dose CQL groups (n=10) respectively. The urate nephropathy model was induced by intragastrically administering adenine and feeding yeast. Distilled water was intragastrically administered at the daily dose of 10 mL/kg to rats in the normal control group and the model group. Allopurinol was intragastrically administered at the daily dose of 9.33 mg/kg to rats in the positive control group. CQL was intragastrically administered at the daily dose of 3.77, 1.89, 0.94 g/kg to rats in the high-, medium-, and low-dose CQL groups. Rats of each group were executed in batches at the 4th and 6th week respectively. Their kidney tissues were taken out to determine the mRNA transcription level of TLR2 and TLR4 by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression level of TLR2 and TLR4 were determined by Western blot. The protein expression level of TLR4 was also detected by immunohistochemical assay.

RESULTSAt week 4 and 6, the protein expression of TLR2 and TLR4 as well as the mRNA transcription of TLR4 increased in the model group, when compared with the control group (P < 0.05, P < 0.01). Compared with the model group, there was no statistical difference in the transcription level of TLR2 mRNA or TLR4 mRNA among the 3 CQL groups (P > 0.05) at week 4 and 6. Additionally, at week 6, the protein expression of TLR4 and TLR2 could be reduced by CQL (P < 0.05, P < 0.01).

CONCLUSIONCQL might protect kidney tissue against inflammatory injury by inhibiting the protein expression levels of TLR2 and TLR4.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; drug effects ; metabolism ; Kidney Diseases ; drug therapy ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Uric Acid

OBJECTIVETo explore the molecular mechanism of exocrine immune inflammatory injury of Sjögren's Syndrome and the intervention of Banxia Qinlian Decoction (BQD).

METHODSTotally 18 female NOD mice were randomly divided into the model group, the positive drug group, and the BQD group, 6 in each group. Six female BALB/c mice were recruited as a blank control group. Mice in the blank control group and the model group were gavaged with deionized water at the daily dose of 0.1 mL/10 g body weight. Tripterygium Tablet was administered by gastrogavage to mice in the positive group at the daily dose of 10 mg/kg. BQD was administered by gastrogavage to mice in the BQD group at the daily dose of 60 g crude drugs/kg. After 12 weeks of medication, mice were sacrificed. Their eyeballs were excised and blood collected. Tissues of bilateral parotids and submandibular glands were kept. mRNA transcriptional levels of IL-17, IL-6, type 3 muscarinic acetylcholine receptors (M3R), aquaporin protein-5 (AQP5) were detected by RT-PCR. Expression levels of M3R and AQP5 protein were detected by Western blot. Protein expression levels of IL-17 and IL-6 were detected by ELISA.

RESULTSCompared with the normal group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly up-regulated in the model group (P < 0.01). Compared with the model group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly down-regulated in the positive drug group and the BQD group with statistical difference (P < 0.01, P < 0.05). Compared with the BQD group, mRNA-transcriptional levels of IL-17, IL-6, and M3R, as well as M3R and AQP5 protein expression levels were significantly down-regulated in the positive drug group (all P < 0.01).

CONCLUSIONThe molecular mechanism of BQD in inhibiting SS exocrine neurotoxic injury might be possibly related to regulating Th17/IL-17 immune inflammatory way.

Animals ; Aquaporin 5 ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Interleukin-17 ; metabolism ; Interleukin-6 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Inbred NOD ; Sjogren's Syndrome ; drug therapy ; immunology ; Submandibular Gland ; Th17 Cells ; Up-Regulation

OBJECTIVESTo investigate mechanism for the increasing level of serum vascular endothelial growth factor(VEGF) in tumour patients during radiotherapy and the inhibitory action of the antisense oligodeoxynucleotide (AS-ODN) to the expression of VEGF protein by radiotherapy in the prostate cancer cell line (PC3M).

METHODSTo observe the changes of serum VEGF in the prostate cancer patients during radiotherapy dynamically and the inhibitory action of the antisense oligodeoxynucleotide to the expression of VEGF by radiotherapy in PC3M.

RESULTSThe changes of serum VEGF in three patients receiving radiotherapy had been observed continuously. The levels of serum VEGF began to increase when the patients received radiotherapy and rised up to peak value after fifteen days, then declined to the range of pre-radiotherapy. Irradiating the PC3M cells with X-rays significantly increased the VEGF expression and secretion. The expression of VEGF protein in the group treated by VEGF AS-ODNs and X-ray irradiation decreased significantly than the group treated only by X-ray irradiation.

CONCLUSIONSThe induction of VEGF protein expression by X-ray irradiation in tumor cells may result in the increasing of the VEGF in the prostate cancer patients during radiotherapy and the induction can be blocked by VEGF AS-ODNs.

DNA, Antisense ; pharmacology ; Endothelial Growth Factors ; antagonists & inhibitors ; blood ; genetics ; Gene Expression ; drug effects ; radiation effects ; Humans ; Intercellular Signaling Peptides and Proteins ; blood ; genetics ; Lymphokines ; antagonists & inhibitors ; blood ; genetics ; Male ; Prostatic Neoplasms ; blood ; pathology ; Radiotherapy ; adverse effects ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo investigate the long-term results of cricohyoidoepiglottopexy(CHEP) in the treatment of glottic carcinoma.

METHODSA retrospective chart review of 92 consecutive patients who underwent CHEP in the selected institut from January 1990 to December 2008. Of the 92 cases, 41 cases of stage I, 39 cases of stage II, 12 cases of stage III. The time for the decannulation and the removal of the nasal feeding and quality of phonation were used for evaluating postoperative functional rehabilitation. The estimated long-term survival rates were calculated by Kaplan-Meier method.

RESULTSAmong 92 patients, 26 cases (27 sides) received neck dissection and lymph nodes were pathologically positive in 3 patients (11.1%). All final surgical margins were negative for tumor invasion. Seven patients had postoperative radiotherapy and one had chemoradiation. Thirteen patients (14.1%) were found local recurrence and nine patients (9.8%) had postoperative metastases. Overall 3-, 5- and 10-year survival rates were 90.0%, 84.5% and 67.0% respectively. Cox multivariate analysis showed that the recurrence had significant influences on the overall survival rate.

CONCLUSIONCHEP not only is relatively easy to master, but also effective in the treatment of glottic carcinoma.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; pathology ; surgery ; Cricoid Cartilage ; surgery ; Epiglottis ; surgery ; Female ; Glottis ; pathology ; Humans ; Laryngeal Neoplasms ; pathology ; surgery ; Laryngectomy ; methods ; Male ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; methods ; Retrospective Studies ; Treatment Outcome

AIM To investigate the effect of β-cyclodextrin-assisted extraction on Cassiae Semen.METHODS The chemical constituents in aqueous extract and β-cyclodextrin extract determined and analyzed by UPLC and principal component analysis had their antioxidant activities tested by six methods (protective effects on lipid peroxidation injuries induced by spontaneous liver,CCl4,H2 O2,FeSO4-vitamin C,and scavenging effects on DPPH,hydroxyl free radicals).RESULTS Compared with aqueous extract,the component content and antioxidant activity of β-cyclodextrin extract were increased by 10.476％ and 80.88％,respectively.CONCLUSION β-Cyclodextrin can effectively enhance the component extraction rate and antioxidant activity of Cassiae Semen.