Objective To investigate the expression and effect of secondary lymphoid tissue chemokine(SLC)in experimental ulcerative colitis(UC)in rats.Methods Thirty Spague-Dawley rats were divided into control group and UC group.SLC expression in colon tissues was detected by RT-PCR and immunohistochemistry,respectively.Results The transcriptional level of SLC in UC tissues was significantly higher than that in the control group(0.846?0.07 vs 0.312?0.12,P＜0.01).The positive expression of SLC was concentrated mainly on submucosa,and the positive rate of SLC protein expression in UC group significantly higher than that in control group(P＜0.01).Conclusion SLC overexpression could contribute to the pathological processes in UC rats,thus SLC may be an ideal ther- apeutic target for UC.

Genetic engineering on macrolide antibiotics was a new field in recent years and more than 100 novel polyketides had been produced until then. Using genomic DNA of S. erythraea A226 as a template, about 3.2 kb DNA fragment without KR6 domain was amplified by overlapping PCR technique and cloned into pWHM3 carrier, which resulted in the construction of homologous recombinant plasmid pWHM2201. Plasmid pWHM2201 was introduced into protoplasts of S. erythraea A226 by PEG-mediated transformation and integrated into the gene locus for erythromycin biosynthesis. After integrants grew for two generations on R3M media without Tsr, they were protoplasted and grown on R3M plates. By PCR identification, 8 mutants without KR6 domain were selected out and named S. erythraea M(1-8). With the identification of mass spectrometry, it was proved that S. erythraea M1 synthesized a novel ketolide compound 3-deoxy-3-oxo-erythronolide B.
Anti-Bacterial Agents ; biosynthesis ; chemistry ; Chromosomes, Bacterial ; Erythromycin ; analogs & derivatives ; biosynthesis ; chemistry ; Genetic Engineering ; Ketolides ; Molecular Structure ; Multienzyme Complexes ; genetics ; Saccharopolyspora ; enzymology ; genetics ; metabolism

OBJECTIVETo investigate the clinical outcomes of anterior corpectomy combined with anterior intervertebral decompression and fusion for multilevel cervical spondylotic myelopathy.

METHODSThe clinical data of 28 patients with multilevel cervical spondylotic myelopathy who underwent surgery from October 2012 to June 2014 were retrospectively analyzed. There were 18 males and 10 females, aged from 45 to 77 years old with an average of (60.11±9.37) years. Three levels were involved in 27 cases, while four levels were involved in 1 case. The preoperative JOA score was 8.89±1.87; the fusion segments angles was (4.87±4.56)°; and the cervical curvature was (11.68±1.25)°. Anterior hybrid decompression and fusion were performed in 28 patients. The fusion segments angles and the cervical curvature were assessed by X-rays at 1, 12 months after operation, respectively. JOA score was used to evaluate the clinical effect.

RESULTSThe operative time was 163 min on average (ranged from 120 to 205 min), and intraoperative bleeding was 198 ml on average(ranged from 100 to 300 ml). Hoarseness occurred in 1 case and got recovery at 3 weeks after operation and choke cough occurred in 1 case, and got improvement at 1 week after operation. All the patients were regularly followed for 12-24 months with an average of(18.46±3.20) months. Graft bone obtained fusion at 12 months after operation and the position of internal fixation was good. The fusion segments angles, the cervical curvature and JOA scores were significantly improved at 1, 12 months after operation(<0.05). The improvement rate of JOA score was(46.46±20.26)% at 12 months after operation, 12 cases got excellent results, 14 good and 2 fair.

CONCLUSIONSAnterior corpectomy combined with anterior intervertebral decompression and fusion is safe and effective and can get satisfactory effects for multilevel cervical spondylotic myelopathy.

OBJECTIVETo investigate the effect of secondary lymphoid tissue chemokine (SLC) on experimental colon lesions in rats with ulcerative colitis.

METHODSSixty Sprague-Dawley rats were randomly divided into control group, model group and SLC intervention group. Colonic mucosal lesions of different groups were observed with HE staining for inflammation and lymphocyte homing situation. Cytokine IL-2 and IL-6 levels were measured by ABC-ELISA. Semi-quantitative RT-PCR was used to examine the colonic SLC expression.

RESULTSIntestinal inflammation score and colonic cytokine levels were significantly different among three groups (P<0.05, P<0.01). Abnormal lymphocyte homing phenomenon under colonic mucosa was found in the model group and the intervention group. SLC mRNA expression of the model and intervention groups increased significantly compared with the control group (0.846+/-0.047, 0.768+/-0.135 vs 0.312+/-0.112, P<0.01). However, there was no significant difference between model group and intervention group.

CONCLUSIONSSLC may play an important role in experimental colonic mucosal inflammation in rats with ulcerative colitis. Blockade of SLC may be one of effective ways in reducing colonic mucosal inflammation.

Animals ; Chemokine CCL21 ; metabolism ; Colitis, Ulcerative ; metabolism ; physiopathology ; Female ; Inflammation ; Interleukin-2 ; metabolism ; Interleukin-6 ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the role of integrin α4β7 in the development of ulcerative colitis (UC) in rats.

METHODSSixty Sprague-Dawley rats were randomly divided into the control group (acetone enema), the model group (2,4-dinitrochlorobenzene, DNCB enema), and the α4 intervention group. Colonic mucosa of different groups was observed and compared in terms of pathology and cytokine changes(IL-2 and IL-6) using ELISA. Semi-quantitative RT-PCR was used to detect the colon α4β7 expression. Integrin α4β7(+) lymphocytes in the portal vein of rats were determined by flow cytometry.

RESULTSThe expression of α4 mRNA was 0.68±0.24 in the model group and 0.58±0.37 in the intervention group, and the expression of β7 mRNA was 0.84±0.37 in the model group and 0.65±0.30 in the intervention group, which were all significantly higher as compared to those in the control group(0.15±0.13 for α4 and 0.24±0.62 for β7, P<0.01). The proportions of integrin α4β7 positive lymphocytes in the portal vein in the model group and intervention group were significantly higher than that in the control group [(76.7±8.2)% and (68.2±7.6)% vs. (14.7±6.7)%, P<0.01]. The expression of IL-2 and IL-6 and the result of macroscopic and microscopic scores in the intervention group were lower than those in the model group(P<0.05).

CONCLUSIONSHigh expression of α4β7 may play an important role in experimental colon mucosa inflammation in rats with ulcerative colitis. The blockade of integrin α4β7 may be a potential target to reduce colonic mucosa inflammation.

Animals ; Colitis, Ulcerative ; metabolism ; pathology ; Colon ; metabolism ; pathology ; Disease Models, Animal ; Female ; Integrins ; metabolism ; physiology ; Interleukin-2 ; metabolism ; Interleukin-6 ; metabolism ; Intestinal Mucosa ; pathology ; Rats

In order to study the relationship between lengths of homologous fragments and chromsomic recombination rate in Saccharopolyspora erythraea, three homologous sequences, with mutant loci and different flanking sequences, (26bp + 27bp), (500bp + 576bp) and (1908bp + 1749bp), were synthesized by chemical reaction or PCR amplification, and cloned into pWHM3 to construct homologous recombination plasmids, pWHM1113, pWHM1116 and pWHM1119. When the plasmids were transformed into protoplast of Saccharopolyspora erythraea A226 under PEG mediated, on an average 30, 69 and 170 transformants grew on each plate for the three plasmids respectively, but chromosomic integration frequency were 0, 2% and 19% among corresponding transformants. Both pWHM1116 and pWHM1119 could take double crossover recombination, and exchange the mutant loci in the chromosome. It was concluded that when the flanking sequences were equal or more than (500bp + 576bp), they could take effective single and double recombination with Saccharopolyspora erythraea chromosome.
Chromosomes, Bacterial ; genetics ; Polymerase Chain Reaction ; Recombination, Genetic ; genetics ; Saccharopolyspora ; genetics

BACKGROUND: Basal metabolic rate (BMR) is an important indicator of human energy metabolism, and low BMR leads to the dysfunction of liver and kidney. Low BMR is usually found in patients with hip fractures, but there is a lack of study on the relationship between mortality of hip fracture and low BMR. OBJECTIVE: To investigate the effect of low BMR on the 1-year mortality in older adults with hip fractures. METHODS: Totally 507 patients with hip fractures aged more than 60 years from January 2014 to March 2016 were included in this retrospective study. Age, sex, surgery or not, surgical pathway, duration from injury to surgery, hospitalized pulmonary infection, number and kind of comorbidities, and 1-year mortality were recorded. BMR on admission was recorded, and multiple Logistic regression analysis was applied. RESULTS AND CONCLUSION: All patients were followed up for 13-15 months, and the 1-year mortality was 13.41% (68/507). The mortality in the low BMR group was significantly higher than that in the non-low BMR group (P ＜ 0.05). Logistic regression analysis showed that older age, conventional treatment, number of combined medical diseases, hospitalized pulmonary infection, and low BMR are risk factors for 1-year mortality in older adults with hip fracture. These results imply that low BMR is strongly associated with 1-year mortality in older adults with hip fracture. BMR can reflect the nutritional status, neuroendocrine, cellular and energy metabolism. Thereafter, for older adults with hip fractures and low BMI, nutrition therapy, re-warming, and endocrine therapy may help reduce the trauma-induced mortality.

OBJECTIVETo compare the short-term efficacy of anterior cervical discectomy and fusion(ACDF) with traditional nail plate system and Zero-profile device in the treatment of cervical spondylotic myelopathy(CSM).

METHODSThe clinical data of 45 patients with CSM treated from July 2014 to August 2015 was retrospectively analyzed. There were 23 males and 22 females with an average age of 53.7 years old(range, 32 to 71 years old). The course of disease was 5 months to 2 years. All the patients were treated with ACDF with 24 cases by traditional nail plate system fixation(group A) and 21 cases by Zero-P system fixation(group B). Operation time and intraoperative bleeding were compared between two groups. Neurological function and cervical pain were evaluated by Japanese Orthopaedic Association scores (JOA) and visual analogue scale (VAS), respectively. Cervical curvature(Cobb angle) change and intervertebral fusion were evaluated by X-rays and CT. And associated complications were analyzed in two groups.

RESULTSAll the patients were followed up for 12 to 16 months with an average of 14 months. Operation time of group A and B was(87.6±23.2) min and (62.7±17.3) min respectively, and the difference was significant between two groups; and intraoperative bleeding was (80.2±36.8) ml and (78.4±29.6) ml respectively, and the difference was not significant. At final follow-up, JOA and VAS of all patients were obvious improved, but there was no significant difference between two groups. Preoperative Cobb angle in group A and B was (8.7±4.3) ° and (8.6±4.2) ° respectively, and the difference was significant. The Cobb angle at final follow-up was (14.5±6.4) ° and (17.4±8.6) ° respectively, and the difference between two groups was significant. The incidence of dysphagia in group A and B were 29.17% and 9.52% respectively, and there was significant difference between two groups. All intervertebral spaces got fusion at final follow-up. No tracheo-asophageal injury and recurrent laryngeal nerve damage or other complications were found. No fusional migration, subsidence, loosening, breakage, etc. were found.

CONCLUSIONSThe clinical comparison of Zero-P interbody fixation system and cervical plate internal fixation for the treatment of cervical spondylosis was quite fair, but Zero-P showed a better therapeutic effect with improvement of life quality.

The Concurrent treatment of the brain and heart (CTBH) theory is proposed based on traditional Chinese medical theory and clinical practice. In this study, a framework for the pharmacological research platform was established to investigate the principles of concurrent treatment of the brain and heart. The platform for CTBH includes several key techniques for network modeling, discovery of active substances, dissecting mechanism of action and investigation of pharmacokinetic property of TCM. Taking network modeling of CTBH as an example, using database search, literature mining, network construction and module analysis, the that network modules closely associated with the pathological progress of cardiovascular and cerebrovascular diseases were identified, while further functional enrichment analysis of these modules indicated that the key biological processes included oxidative stress, metabolism and inflammation. GSK3B, NOTCH1, CDK4 were identified as key nodes in these network modules. The above-mentioned platform was applied to construct component-biomolecules network of Danhong injection for the identification of common targets and pathways. Among them, GSK3B had the highest correlation with the composition of Danhong injection in the network, and the biological function of whose cluster was related to cell oxidative stress. Based upon results of network analysis, validation experiments suggested that Danhong injection significantly improved the survival rate of oxidative injured myocardial cells and nerve cells, and the protective effect was related to the increase of phosphorylated GSK3β protein expression. Moreover, extracts of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos exerted the synergisticcytoprotective effect. The results indicated that the mechanism of treatment of cardiovascular and cerebrovascular diseases of Danhong injection could be studied through network modeling and other methods. In summary, the proposed pharmacological platform provided a feasible way for revealing the mechanism of CTBH by using modern scientific methods.

Objective:To explore the antidepressant mechanism of Yinxing Mihuan oral solution （YMO） by investigating its effect on depression model rats. Method:The depression rats were induced by isolation combined with chronic unpredictable mild stress （CUMS） and then randomly divided into model group， fluoxetine group （10 mg·kg^-1） and high-dose （618 mg·kg^-1） and low-dose （309 mg·kg^-1） YMO groups. A blank control group was also set up and ten rats were included in each group. Modeling lasted for 21 consecutive days， and rats were administered the 8th day after stimulation at a dose of 10 mL·kg^-1 for 14 days， except those in the blank control and model groups which were given distilled water. Afterward， the sucrose preference test， open field test， tail suspension test were carried out. The pathological changes of hippocampus in depression rats were observed after hematoxylin-eosin （HE） staining. The content of interleukin-1β （IL-1β）， interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in the hippocampus of rats in each group and the expression of NOD-like receptor 3 （NLRP3） and other proteins in its related activation signaling pathways were detected with multi-factor detection （Luminex） and Western blot. Result:After 14 days of continuous administration， compared with the blank control group， the model group witnessed significantly reduced sugar water consumption rate and the times of rearing and significantly prolonged cumulative time of immobility during tail suspension （P<0.05， P<0.01）. Compared with the model group， the fluoxetine group and the high-dose YMO group saw increases in the times of rearing， times of crossing and sugar water consumption rate and a significant decrease in the cumulative time of immobility during tail suspension （P<0.05， P<0.01）. The results of HE staining showed that the neurons in the hippocampus of rats in the high-dose YMO group were arranged in order and slightly loosened， without obvious microglia infiltration observed. The levels of IL-1β， IL-6 and TNF-α in the hippocampus of the model group increased significantly as compared with the blank control group （P<0.05， P<0.01）， and their content in the high-dose YMO group was significantly lowered in the comparison with the model group （P<0.05， P<0.01）. Molecular biology experiments demonstrated that compared with the results of blank group， the expression of purinergic receptor P2X7 （P2RX7）， NLRP3， apoptosis-associated speck-like protein （ASC）， Caspase-1 and IL-1β remarkably increased in the model group （P<0.05， P<0.01）. Additionally， the expression of P2RX7， NLRP3， ASC， Caspase-1 and IL-1β was significantly inhibited in the fluoxetine group and the high-dose YMO group compared with the model group （P<0.05， P<0.01）. Conclusion:YMO can improve the depression-like behaviors of rats induced by isolation combined with CUMS， and its mechanism of action is related to the regulation of the P2RX7/NLRP3 signaling pathway.