Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-kappaB and the expression of chemokine MOB-1, MCP-lgenes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-kappaB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10(-3) mol/L, 10(-4) mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10(-3) mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-kappaB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10(-3) mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10(-5) mol/L atropine) or NF-kappaB inhibitor (10(-2) mol/L PDTC) could obviously inhibit the activation of NF-kappaB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-lgenes in pancreatic acinar cells in vitro through the activation of NF-kappaB.

Objective To describe the technique and therapeutic effect of modified unilateral laminotomy for bilateral decompression (M-ULBD) for lumbar spinal stenosis (LSS).Methods A total of 56 patients with LSS were randomly divided into group A and B.The 27 patients in group A (15 males and 12 females,with an average age of 59.4 years) underwent M-ULBD.The other 29 patients in group B (18 males and 11 females,with an average age of 61.6 years) received conventional laminectomy.JOA score of low back pain,VAS,CPK three days after operation,pre-and post-operative cross-sectional areas of multifidus were used to evaluate the clinical results.Results A total of 45 patients (21 in group A and 24 in group B) completed 2 years of follow-up.The preoperative VAS of low back pain,leg pain,numbness,JOA score and cross-sectional areas of multifidus were 5.6±1.7,7.1±0.4,11.6±2.6,5.8±1.8 cm2 in group A and 6.2±1.2,7.9±1.3,10.9±1.0,6.1±2.0 cm2 in group B.There was no significant difference in preoperative data between both groups.The union of split spinous process was observed in all cases 6months later according to computed tomography.The postoperative CPK was lower in group A.The postoperative JOA and VAS scores in both groups were improved significantly compared with the corresponding preoperative ones.The VAS of leg pain,numbness,JOA score,and JOA recover rate in latest follow-up were 1.3±0.2,1.5±0.7,26.7±2.1,86.1％±3.1％ in group A,and 1.7±0.3,2.0±1.3,24.3±2.5,83.6％±6.4％ in group B,respectively.All these data have no difference between group A and B.The VAS of low back pain and atrophy rate of multifidus were 1.0±0.5,6.4％±1.2％ in group A,and 2.6±0.7,15.7％±3.0％ in group B respectively.All these data are lower in group A.Conclusion Our two years follow-up shows that this method is efficient for lumbar spinal stenosis treatment,however,it still need long term follow-up and to compare with other modified methods.

OBJECTIVETo explore the clinical effect of the sacrococcygeal space injection for the treatment of failed back surgery syndrome.

METHODSFrom July 1998 to October 2012,47 patients with failed back surgery syndrome were treated and included 39 males and 8 females with an average age of 61.5 years old ranging from 35 to 89 years old. Among them,41 patients experienced one time of operation, 6 patients with twice of operation. Forty-one patients underwent single,bilateral fenestration or central laminectomy decompression, discectomy. Six patients underwent total laminectomy discectomy and inter body fusion and pedicle screw fixation. All patients were examined by X-ray plain film, CT or MRI before treatment. The anticoagulation was discontinuation before treatment. The needle was put into the sacrococcygeal gap at prone position in the sense of frustration,suction without cerebrospinal fluid and blood,with injection of Mailuoning (Chinese characters: see text) 15 ml. The pain was assessed by VAS before and after treatment. The Oswestry low back pain disability index and survival quality interference degree were evaluated.

RESULTSAt 1 month after treatment,the pain VAS decreased from 59.24 +/- 17.35 before treatment to 19.19 +/- 11.19 after treatment (P < 0.05); The Oswestry low back pain disability index decreased from (41.35 +/- 9.87)% before treatment to (23.17 +/- 17.56)% after treatment (P < 0.05); The survival quality interference degree decreased from 6.5 +/- 2.2 before treatment to 2.6 +/- 1.4 after treatment (P < 0.05).

CONCLUSIONThe sacrococcygeal gap injection for treatment of failed back surgery syndrome has advantages of simple, safe, fewer complications, and low treatment cost.

Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; administration & dosage ; Failed Back Surgery Syndrome ; diagnostic imaging ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Radiography ; Sacrococcygeal Region ; diagnostic imaging

OBJECTIVETo observe the clinical effect of chronic fatigue syndrome (CFS) treated with moxibustion at Gaohuang (BL 43).

METHODSWith stratified block randomization, 72 patients accorded with inclusive criteria were divided into a moxibustion at Gaohuang (BL 43) group (moxibustion group) and an acupuncture group, 36 cases in each one. In the moxibustion group, Gaohuang (BL 43) was treated with big moxa cones as the main acupoint, 10 cones a time; Qihai (CV 6) and Zusanli (ST 36) were added with big moxa cones, 7 cones a time. In the acupuncture group, acupoints were the same as those in the moxibustion group, and twirling reinforcing method was used after qi arriving, 60 times one minute and 360° with range. In the two groups, 10-day treatment was made into one course and there were two days between courses. The treatment was given once a day for 3 courses. Changes of fatigue assessment index (FAI) before and after treatment and clinical effects were observed.

RESULTSThe total effective rate was 88.9% (32/36) in the moxibustion group, which was better than 72.2% (26/36) in the acupuncture group apparently (P < 0.05). After treatment in the two groups, FAI scores were obviously declined compared with those before treatment (both P < 0.01) and FAI score in the moxibustion group was apparently lower than that in the acupuncture group (P < 0.05).

CONCLUSIONMoxibustion at Gaohuang (BL 43) can improve the FAI score of patients with CFS and the clinical efficacy is definite.

Acupuncture Points ; Adult ; Fatigue Syndrome, Chronic ; therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the effect and mechanism of hirudin on atherosclerotic plaques in apolipoprotein E knockout (ApoE(-/-)) mice.

METHODSTotally 24 ApoE(-/-) mice, 7-8 weeks old were fed with high fat diets. They were randomly divided into the recombinant hirudin treatment group (drug group) and the model group according to body weight and different dens, 12 in each group. Twelve C57BL/6J mice, 7-8 weeks old fed with high fat diet were recruited as the normal control group. Recombinant hirudin (0.25 mg/kg) was intraperitoneally injected to mice in the drug group from the 10th week old once every other day for five successive weeks. Equal volume of normal saline was injected to mice in the model group. Mice in the normal control group received no treatment. All mice were sacrificed after fed with high fat diet until they were 20 weeks old. Serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), high-sensitive C-reactive protein (hs-CRP), E-selectin, interleukin-6 (IL-6), and stromal metalloproteinase-2 (MMP-2) were detected. The plaque/lumen area and extracellular lipid composition/ plaque area were analyzed by HE staining and morphometry. Changes of signaling molecules in store-operated calcium channels, including stromal interacting molecule 1 (STIM1), Orail protein, and transient receptor potential channel 1 (TRPC1) were determined by Western blot. Results Lipid plaque formed in the aorta vessel wall of 20-week old mice in the model group. Compared with the normal control group, serum levels of TC, TG and LDL increased (P<0.01), hs-CRP, E-selction, IL-6, and MMP-2 obviously increased (P<0.01, P<0.05) in the model group; expression levels of STIM1, TRPC1, and Orail significantly increased (P<0.01). Compared with the model group, the plaque/lumen area and the extracellular lipid composition/plaque area significantly decreased in the drug group (P<0.05, P<0.01); serum levels of TC and LDL, hs-CRP, E-selction, IL-6, and MMP-2 obviously decreased (P<0.05, P<0.01); expression levels of STIM1, TRPC1, and Orail were significantly down-regulated (P<0.05, P<0.01).

CONCLUSIONHirudin could significantly improve lipids and endothelial functions of ApoE(-/-) mice, down-regulate expression levels of STIM1, Orai1, and TRPC1, and thus delaying the occurrence and development of atherosclerosis.

Animals ; Aorta ; Apolipoproteins E ; metabolism ; Atherosclerosis ; C-Reactive Protein ; Cholesterol ; Diet, High-Fat ; Drugs, Chinese Herbal ; E-Selectin ; Hirudins ; metabolism ; Interleukin-6 ; Lipids ; Lipoproteins, HDL ; Lipoproteins, LDL ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Plaque, Atherosclerotic ; metabolism ; Recombinant Proteins ; metabolism ; Triglycerides

OBJECTIVE: To investigate the effect and mechanism of tea polyphenol (TP) on the proliferation, apoptosis, migration and invasion of nasopharyngeal carcinoma(NPC) cell line HONEl. METHOD: After treated with different concentration of tea polyphenol, CCK-8 assay, fluorescent staining, cell scratching assay and transwell assay were applied to detect the effect of tea polyphenol on the HONE1 cells. Furthermore, the expression of protein VEGF was investigated by flow cytometry assay. RESULT: It was found that tea polyphenol could inhibit NPC cell proliferation significantly in a dose-dependent manner, however, little impact was observed in normal nasopharyngeal epithelial cell line NP69. Furthermore, it was demonstrated by fluorescent staining assay that tea polyphenol could induce NPC cell apoptosis, and cell scratching assay and transwell assay showed that tea polyphenol could inhibit cell migration and invasion. CONCLUSION Tea polyphenol can significantly inhibit cell proliferation, induce cell apoptosis and decreased the migration and invasion ability of NPC cells in vitro. Tea polyphenol might be a tumor suppressor of NPC cells.
Apoptosis ; drug effects ; Carcinoma ; Cell Line, Tumor ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; pathology ; Polyphenols ; pharmacology ; Tea ; chemistry

Objective To review and analyze the clinical effect of combined posterior mini-invasive fixation with anterior debridement via small incision for the treatment of single segment lumbar vertebral tuberculosis. Methods Totally 31 cases with single segment lumbar tuberculosis (both borderline tuberculosis) without attachment involvement underwent one-stage anterior debridement, interbody fusion and posterior mini-invasive fixation from July 2010 to July 2015. Among these patients, 19 were male and 12 were female. The average age was (36.1±17.8) years old (ranged 21-61 years old). The average course of disease was 11(9, 12) months (ranged from 2 to 16 months). All were single segment involvement, and the involved segment was L2-3 in 7 cases, L3-4 in 10 cases, L4-5 in 6 cases, and L5-S1 in 8 cases. The clinical manifestations included lumbar back pain in 31 cases with an average pain visual analog score (VAS) of 7(6, 8) points. ASIA grade of spinal cord injury was E in 25 cases and D in 6 cases. Paravertebral abscess occurred in 22 cases and iliac fossa gravity abscess appeared in 9 cases. Kyphosis was observed in 12 cases and the average Cobb angle was 21° ± 6° . Quadruple anti-tuberculosis chemotherapy was used for at least 2 weeks preoperatively. Posterior mini-invasive fixation was fulfilled on prone position, including mini-invasive percutaneous screws in 18 cases and pedicle screw fixation via Wiltse approach in 13 cases. Posterior distraction and deformity correction were performed simultaneously for patients with kyphosis. Then the patients were changed to lateral position for anterior debridement, bone grafting and/or titanium mesh fusion. Results The average operation time was (204±54) min (ranged 160-240 min) in 31 patients, and the mean blood loss was (168±73) mL (ranged 100-300 mL). Delayed healing of anterior incision occurred in 1 case and the incision healed after two-week dressing of wound. The incision healed well in the rest 30 cases. No complications such as nerve function, blood vessel injury were found in patients. The VAS scores of the 3 days after operation were 1.3 ± 0.3 and 2.1 ± 1.4 in percutaneous group and Wiltse approach group, respectively, and the difference between them was statistically significant ( P<0.05). The VAS score of low back pain was 2(1, 3) points in all the 31 patients three months after operation, which was significantly lower than that before surgery (P<0.05). The six patients with neurological symptoms recovered to E grade after operation. The average Cobb angle correction was 15°±5° in 12 patients with kyphosis (P<0.05), which was significantly decreased compared with that before surgery (P<0.05). All patients were followed up for an average of (36.8 ± 9.3) months (ranged from 12 to 72 months). The clinical healing of tuberculosis was achieved at the final follow-up in all the 31 patients. No complications were observed, such as lumbar kyphosis, internal fixation loosening and breakage, dislocation and titanium mesh subsidence. Conclusion Mini-invasive posterior internal fixation and anterior debridement via small incision is effective for the treatment of single segment lumbar vertebral tuberculosis in lesion debridement and spine stability reconstruction by short segment fixation. This technique can reduce fused segments, surgical trauma of anterior approach and related complications.

Objective To explore Toll like receptor 2 (TLR2) expression after partial liver ischemia/ reperfusion (I/R) in BALB/c mice and its relationship with liver function impairment. Methods All the animals were randomly divided into ischemia/reperfusion injury (I/R) and sham operation (S) groups. Total RNA was extracted from the liver samples and the expression of TLR2 mRNA was analyzed quantitatively by real time PCR method. Membrane protein was extracted, and the expression of TLR2 protein was detected by western blot method. Portal vein serum and plasma were taken for further detection of the levels of alanine aminotransferase (ALT), tumor necrosis factor alpha (TNF ?) and endotoxin . Results After 1h of partial liver ischemia and 4 h of reperfusion, the expression of TLR2 mRNA and TLR2 protein were both remarkably upregulated in tissues of ischemic liver lobes in I/R group compared with that in S group (value of TLR2mRNA: 1.06?0.91vs5.08?1.32,P0.05). Conclusions TLR2mRNA and TLR2 protein expression were upregulated in tissues of ischemic liver lobes after partial I/R injury in mice, and were associated with increased levels of portal vein ALT and TNF ?,and impairment of liver function.

AIM: To construct prokaryotic expression vector of human angiogenesis inhibitor arresten gene and express recombinant arresten in Escherichia coli. METHODS: Human arresten gene was amplified from recombinant plasmid pGEM-Arr with polymerase chain reaction (PCR), and then cloned into prokaryotic expression vector pRSET by means of recombinant gene technology. The recombinant plasmid pRSET-Arr was transformed into E.coli BL21(DE3), and recombinant arresten was expressed in the bacteria under induction of IPTG. The expressed products were detected by SDS-PAGE analysis. RESULTS: Restriction analysis indicated that the arresten gene was successfully inserted into the expression vector, and DNA sequencing verified that the reading frame of the recombinant vector was correct. Recombinant arresten was successfully expressed in Escherichia coli; its molecular weight was about 26 kD and its amount was approximately 30% of total bacterial proteins.CONCLUSION: The successful construction of prokaryotic expression vector containing human arresten gene and the effective expression of recombinant arresten in Escherichia coli laid the foundation for further study on its biological functions.

Objective To assess the psychometric properties of the Chinese Heartburn Version of Quality of Life in Reflux and Dyspepsia Questionnaire (QOLRAD) in patients with gastroesophageal reflux disease.Methods 130 patients with symptoms of heartburn completed the Chinese version of QOLRAD,the Short-Form-36 (SF-36).30 of them received proton pump inhibitors (PPI) for 8 weeks,which was used to test responsiveness of the Chinese heartburn version of QOLRAD.Results The Chinese version of QOLRAD had acceptable internal consistency.The overall Cronbach's alpha was 0.89 and the internal consistency of dimensions ranged from 0.70～0.90.Content validity index (CVI) was 0.82.Confirmation factor analysis revealed a 5 factor solutions accounting for 62.02％ and most of items in their dimensions had acceptable loads (＞0.4).There was acceptable concurrent validity with correlations between the Chinese heartburn version of QOLRAD and Short Form-36 health survey ranging from 0.172～0.613.As to responsiveness,after therapy of PPI for 8 weeks,except the dimension of sleep disturbance,scores for dimensions of vitality,food/drink problems,physical/social functioning,emotional distress had significant changes.Conclusions The Chinese version of QOLRAD has a good reliability,validity and responsiveness to therapy,which can be used to assess the quality of life in patients with gastroesophageal reflux disease.