ObjectiveTo investigate the relationship between the degenerative mechanisms of lumbar intervertebral disc (LID) and apoptosis.MethodsThe total RNAs were isolated from human LID tissues. Both the mRNAs from the degeneration and normal LID were reversely transcribed to the cDNAs. The cDNAs were labeled with the incorporations of fluorescent dUTP, for preparing the hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and analyzed by computer image analysis. The apoptotic status and the expression of Bcl-2 and Bax in 12 cases of degenerative LID and 10 cases of normal LID were detected with TdT-mediated dUTP-biotin nick end labeling (TUNEL) and immunohistochemistry methods.ResultsAmong the 4096 targets, there were 10 genes related to apoptosis. The expression related to Bax protein gene was up-regulated and it was down-regulated for Bcl-2 protein. In group of normal LID, the average apoptotic index (AI) was (24.897±3.620); percentage of Bcl-2 positive cells was (31.440±4.150)%; percentage of Bax positive cells was (29.372±2.588)%, average optical density (OD) values of positive particles were (0.183± 0.010 ), ( 0.203 ±0.012) and (0.169±0.005) respectively. In group of degenerative LID, the average AI was (49.232±3.440); percentage of Bcl-2 positive cells was (18.239±2.470)%; percentage of Bax positive cells was (52.349±3.764)%; average OD values of positive particles were (0.152±0.003), (0.310±0.008) and (0.262±0.014) respectively. There were significantly differences in AI and expressions of Bcl-2 and Bax proteins between normal LID and degenerative LID (P<0.05).ConclusionCell apoptosis plays an important role in the process of LID degeneration. Both Bcl-2 and Bax take part in the occurrence and progression of LID.

Objective To evaluate laparoscopic radical resection of metachronous colorectal carcinoma.Methods A total of 13 patients with metachronous colorectal carcinoma undergoing laparoscopic resection in Department of Gastrointestinal Surgery,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology from January 2013 to December 2015 were analyzed retrospectively.Results The mean time of surgery was (156 ± 9) min.Tumors were located in the right hemicolon in 3 cases,in the transverse colon in one,in the left hemicolon in 2,in the sigmoid colon in four and in the rectum in 4.The mean blood loss was (66 ± 21) ml.There was no conversion to open surgery.Two patients were done with protective ileostomy.Postoperative gastrointestinal function recovery time was (2.5 ± 0.7) days.One postoperative intra-abdominal bleeding was successfully controlled laparoscopically.Posteperative length of hospital stay was (26.2 ± 2.9) days.The median follow-up was 12 months (5-30 months) with no cancer recurrence.Conclusions Laparoscopic radical resection of metachronous colorectal carcinoma has good curative effect,and high success rate in spite of previous history of laparotomy.

Objective: To explore the mechanism of acupuncture in regulating cognitive deficits in insomnia rats by observing the effect of acupuncture on microglia in thalamic reticular nucleus (TRN). Methods: Thirty rats were randomly divided into a control group, a model group and an acupuncture group, with 10 rats in each group. The insomnia model was established by intraperitoneal injection of para-chlorophenylalanine (PCPA) once a day for 2 d. Rats in the control group were intraperitoneally injected with the same amount of normal saline. Rats in the acupuncture group received acupuncture at Neiguan (PC 6) and Zusanli (ST 36) for 5 consecutive days. The CLOCKLAB 2 data acquisition system was used to dynamically observe the sleep of the rats throughout the experiment. The cognition of rats was evaluated by event-related potentials (ERPs). After intervention, brain tissue was extracted. Immunofluorescence was used to test the fluorescence expression in TRN region. The concentrations of interleukin (IL)-1β and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay. Results: After intraperitoneal injection of PCPA suspension, the spontaneous activity in light period of rats in the model group and acupuncture group increased significantly compared with the control group (both P<0.01). After acupuncture treatment, the rats in the acupuncture group had much less spontaneous activity during the light period than those in the model group (P<0.01), and the results indicated that acupuncture could effectively improve the sleep quality of insomnia rats. Compared with the control group, rats in the model group showed that the P3 latency, the average optical density of microglia, and the concentrations of IL-1β and TNF-α increased significantly (all P<0.05), and the P3 amplitude decreased significantly (P<0.01). Compared with the model group, rats in the acupuncture group presented that the P3 latency, the average optical density of microglia, and the concentrations of IL-1β and TNF-α were significantly decreased (all P<0.05), and the amplitude of P3 was significantly increased (P<0.05). Conclusion: Acupuncture possesses an ability to improve the cognitive state in insomnia rats. The mechanism may be related to inhibiting the microglial activation, diminishing the levels of pro-inflammatory mediators like IL-1β and TNF-α, and promoting the recovery of central nervous system function.

Objective To investigate the effect of the polymorphisms of multidrug resistance 1 (MDR1) C3435T and G2677T on Tacrolimus (Tac) individualized treatment and prognosis of grafts in the renal transplantation recipients (RTRs).Methods One hundred and twenty-seven RTRs who treated with Tac regimen and had a stable graft function were enrolled,and were divided into adjuvant treatment group and non-adjuvant treatment group according to whether given adjuvant drugs to raise Tac trough concentrations. MDR1 C3435T and G2677T SNPs were detected by using sequence specific primers PCR.Tac trough concentrations of whole blood were measured by using enzymelabeled immunosorbent assay.Tac concentration-to-dose ratio (C/D) standardized by body weight was compared according to the various genotypes and haplotypes of MDR1 C3435T and G2677TA SNPs.Results Adjuvant treatment group including 36 recipients had a higher frequency of C genotype of C3435T than un-adjuvant treatment group (68.05％ vs 48.35％,P ＜ 0.01 ). The frequency of G2677TA polymorphisms was of no significant difference between the two group recipients (P＞ 0.05).As to non-adjuvant treatment recipients,the mean Tac DD required and C/D were not significantly different among various polymorphisms of MDR1 G2677T/A and C3435T or various haplotypes (P＞0.05).During A follow-up period of 4 years,13 recipients suffered graft dysfunction in which 84.6％ (11/13) carried 3435C genotype (P＞0.05).Conclusion The frequency of MDR1 C3435T polymorphisms in RTRs is high in the recipients given adjuvant treatment to raise Tac concentrations.Recipients with 3435C genotype were prone to graft dysfunction.

To develop an oral drug,ITF gene encoding ITF proein,was expressed in a live delivery vehicle lactococcus lactis.First,the ITF gene was cloned into the prokaryotic expressive vector pNICE:sec.Second,the recombinant vector pNICE:sec-ITF was transformated into Lactococcus lactis strain NZ9000 to express ITF protein.Then the recombinant ITF was induced to express and was identified by SDS-PAGE and Western blot.Rabbits are divided into blank control group,preparation group and therapeutic group which are respectively administrated wih PBS and pNICE:sec-ITF Lactococcus lactis.By grades of ulcer test whether administrated pNICE:sec-ITF Lactococcus lactis protects against HCl-induced gastric injury in rabbits.The results were described as follows.The ITF was amplified and cloned in the vector pNICE:sec successfully.The fusion protein(5.9kDa) was expressed in L.lactis by the induction of the nisin.The quantity of expression accounted for 5% of the total bacterial protein.Western bolt analysis confirmed that fusion protein could be recognized specially by Monoclonal Anti-human TFF3 Antibody.Preparation groups and therapeutic groups do good than control group.Prove that administrated pNICE:sec-ITF Lactococcus lactis is biologically active in an HCl-induced rabbit gastric mucosal injury model.

Objective:To investigate the effects of estradiol on ~(60)Co?-ray induced apoptosis of bone marrow hematopoietic cells of mice,and to discuss the related anti-irradiation mechanism.Methods:KM mice were randomly divided into 3 groups(15 mice/each group):control group(without radiation),pure radiation group and estradiol+radiation group(ER group).The pure radiation group was irradiated by 4.0 Gy?-ray at a dose rate of 1.15Gy/min;the ER group was administered with 0.1 mg estradiol(IM)at 10 days before 4.0 Gy?-ray radiation;and the control group received no special treatment.The apoptotic DNA segments of bone marrow hematopoietic cells were analyzed by DNA agarose gel electrophoresis;flow cytometry was used to examine the apoptosis rate of cells and expression of Fas and Bcl-2 at 4 h,8 h,and 12 h after irradiation.Results:Eight hours after radiation,the apoptotic DNA segments were obviously increased and apoptotic DNA ladder appeared,which was not seen in the other 2 groups.The apoptosis rate of bone marrow hematopoietic cells in ER group was significantly lower than that in the pure radiation group at 4,8,and 12 h after irradiation(P

S1 nuclease (from Aspergillus oryzae) is a specific enzyme to degrade single stranded DNA or RNA molecules. It has been reported to be able to convert superhelical circular DNA molecules into open circle or linear forms under certain conditions, but this function has not been well explored. In order to use the action of S1 nuclease to linearize circular DNA and develop a novel way of cloning microcircular DNAs, the pUC19 was used to investigate the relationship between the linearization efficiency of S1 nuclease and the amount of enzyme used. By this way the optimal conditions for linearization of circular DNAs by S1 nuclease would be determined. 0.3u to 17u S1 nuclease per 100ng pUC19 DNA was added into a 25 microL system, respectively, to perform the reaction. The effectiveness of enzyme digestion was realized by electrophoresis in a 1.2% agarose gel. The results showed that along with the increase in enzyme amount from 0.3u to 17u a gradual decrease in the superhelical form, a gradual increase in the linear form and then in the circular form was obvious. The conversion from superhelical form to linear and circular form was directly related to the enzyme amount used. A higher proportion of linear DNA molecules was achieved by using 5 to 17u S1 nuclease per 100ng DNA. Besides, electrophoretic mobility of the S1 nuclease-linearized pUC19 was the same as that of the linear form produced by restriction enzyme digestion. According to the result of phiX174 digested by S1 nuclease it has been proposed that the enzyme cleaves first randomly on one site of one strand, thus converting the superhelical molecules into open circle form, and then on the same site of the complementary strand to produce the linear form. Therefore, the S1 nuclease-linearized DNA molecules are intact in the sense of their length and can be used for cloning. The plasmid-like DNA pC3 from cucumber mitochondria is a double stranded circular DNA molecule with about 550bp and the smallest known plasmid-like DNA in eukaryotic mitochondria. Many attempts have been made to linearize the molecule by using restriction enzymes but failed. Therefore, S1 nuclease was used to linearize pC3 based on the results obtained with pUC19. The linearized pC3 DNA molecules formed a very sharp band in a 2.5% agarose gel after electrophoresis. They were then recovered from the gel, added an "A" tail and ligated with T-vector. After transformation into E. coli JM109 cells, the positive clones were, screened by the blue-white selection. The insert was then cut using restriction enzymes EcoRI and Pst I. The result of electrophoresis shows that the electrophoretic mobility of the insert is just the same as that predicted. A 32 P-labled probe was synthesized using pC3 as the template and Southern blot analysis was carried out. The result shows that the inserted DNA is hybridized to the probe, which indicates that the cloned DNA fragment is from pC3. The sequence information of the insert shows that the plasmid-like DNA pC3 was 537bp in length. The nucleotide sequence was deposited in the GenBank (the accession number is AF522195).
Blotting, Southern ; Cloning, Molecular ; methods ; DNA, Circular ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Single-Strand Specific DNA and RNA Endonucleases ; genetics ; metabolism

OBJECTIVETo explore the clinical effect of intertransverse approach microendoscopic discectomy for far lateral lumbar disc herniation.

METHODSFrom February 2005 to February 2010, 73 patients with far lateral lumbar disc herniation were treated with intertransverse approach microendoscopic discectomy. Their clinical data were retrospectively analyzed. There were 41 males and 32 females, aged from 19 to 80 years old with an average of 56.5 years; courses of disease ranged from 1 to 25 months with an average of 4.5 months. The main symptom was low back pain and sciatica, especially the sciatica was seriously. Herniation level was in L3,4 of 9 cases, L4,5 of 49 cases, L5S1 of 15 cases. Preoperative, 2 weeks after operation, final follow-up, conditions of pain relief were assessed by visual analogue scale (VAS); total life quality of patients were evaluated by Oswestry Disability Index (ODI) before operation and last follow-up.

RESULTSAll operations were performed successfully, operative time was from 40 to 115 min (mean of 50 min); and blood loss was from 50 to 150 ml (mean of 110 ml). Incision infection had 1 case and incomplete nerve root injury had 1 case. All patients were followed up from 3 to 8 years with an average of 4.5 years. Postoperative VAS and ODI had obviously improved (P < 0.01).

CONCLUSIONThe technique of intertransverse approach microendoscopic discectomy is a feasible and effective method for far lateral lumbar disc herniation.

Adult ; Aged ; Aged, 80 and over ; Diskectomy ; methods ; Endoscopy ; methods ; Female ; Humans ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

AIM To explore the clinical effects of San'ao Decoction combined with salbutamol on patients with cough variant asthma.METHODS Eighty patients were randomly assigned into control group(40 cases)for 3-month administration of salbutamol,and observation group(40 cases)for 3-month administration of both San'ao Decoction and mifepristone.The changes in clinical efficacy,TCM syndrome scores,serum MMP-9,VEGF and sex hormones(E2,FSH,PRL)and incidence of adverse reactions were detected.The changes in clinical efficacy,EOS,SIgG4,TIgE,Eotaxin,NKA,LTD4,IL-27,SP,R5-R20,Fres,R5,Zrs,PEF diurnal variation rate,FEF25,PEF,ACT score,PAQLQ score,recurrence rate and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P<0.05),along with lower recurrence rate(P<0.05).After the treatment,the two groups displayed decreased EOS,TIgE,Eotaxin,NKA,LTD4,SP,R5-R20,Fres,R5,Zrs and PEF diurnal variation rate(P<0.05),and increased SIgG4,IL-27,FEF25,PEF,ACT score,PAQLQ score(P<0.05),especially for the observation group(P<0.05).No significant difference in incidence of adverse reactions was found between the two groups(P>0.05).CONCLUSION For the patients with cough variant asthma,San'ao Decoction combined with salbutamol can improve EOS,TIgE,Eotaxin,SIgG4 levels and airway resistance indices,lung function indices,regulate neurogenic mediators,alleviate airway inflammation injury,enhance life quality,and reduce recurrence rate.

To study the chemical constituents of Asarum himalaicum, fifteen compounds were isolated from a 70% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, and HR-ESI-MS, these compounds were identified as 4-demethoxyaristolochic acid BII (1), aristolochic acid I (2), aristolochic acid Ia (3), 7-hydroxyaristolochic acid I (4), aristolochic acid IV (5), aristolic acid II (6), debilic acid (7), aristololactam I (8), 9-hydroxyaristololactam I (9), 7-methoxyaristololactam IV (10), (2S)-narigenin-5, 7-di-O-beta-D-pyranosylglucoside (11), 4-hydroxybenzoic acid (12), 3, 4-dihydroxybenzoic acid (13), 4-hydroxycinnamic acid (14), and beta-sitosterol (15). All of these compounds (1-15) were obtained from A. himalaicum for the first time. Among them, 1 was identified as a new compound, and compounds 3-6, 9, 12-14 were isolated from Asarum genus for the first time. Since the kidney toxicity of aristolochic acids and aristololactams has been reported, the result of this investigation suggests that it should be cautioned to use A. himalaicum as a medicine.
Aristolochic Acids ; chemistry ; isolation & purification ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Coumaric Acids ; chemistry ; isolation & purification ; Hydroxybenzoates ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Parabens ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Propionates ; Sitosterols ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization