OBJECTIVETo compare the difference in the efficacy on thoracic facet joint disorder between the combined therapy of electroacupuncture and manual reduction and the simple manual reduction.

METHODSOne hundred and sixty patients were randomized into an electroacupuncture and manual manipulation group (group A) and a simple manual manipulation group (group B), 80 cases in each one. In the group A, Ashi points and three pairs of Jiaji (EX-B 2) bilateral to the painful sites were selected. The perpendicular puncture was used at Ashi points, the oblique puncture was used at Jiaji (EX-B 2) and connected with electric stimulation for 20 min, additionally, the corresponding manual reduction was adopted at the sites of facet joint disorder. In the group B, the simple manual reduction was applied to the affected sites. Acupuncture was given once every day, the manual reduction was applied once every 10 days. The treatment of 10 days made one session. The efficacy was analyzed statistically at the end of two sessions of treatment. Before and after treatment, McGill pain scale was adopted for the value statistical analysis. PRI score, VAS score and PPI score of patients were calculated before and after treatment and compared in the two groups. The efficacy was compared between the two groups.

RESULTSThe curative rate was 56.3% (45/80) in the group A, which was better than 18.8% (15/80) in the group B (P< 0.01). The total effective rate was 95.0% (76/80) in the group A, which was better than 76.3% (61/80) in the group B (P<0.01). The scores of PRI, VAS and PPI after treatment were all improved significantly in the two groups (all P<0.05), in which, the results in the group A were better than those in the group B (PRI: 4.00 +/- 0.97 vs 5.44 +/- 1.16, VAS: 3.29 +/- 0.72 vs 3.87 +/- 0.81, PPI: 1.07 +/- 0.74 vs 1.64 +/- 0.90, all P<0.05).

CONCLUSIONThe combined therapy of electroacupuncture and manual manipulation achieves the superior efficacy on thoracic facet joint disorder as compared with the simple manual manipulation. The combined therapy relieves the symptoms of thoracic facet joint disorder and reduces the severity of disorder.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Thoracic Diseases ; therapy

AIM　To investigate the poteintation of vincristine-induecd apoptosis by tetrandrine, neferine and dauricine isolated from Chinese medicinal plants in the human mammary MCF-7 multidrug resistant cells. METHODS　The apoptotic cells were detected by fluorescent staining of a combination of Hoechst 33342 and propidium iodide (PI), flow cytometry and agarose electrophoresis. RESULTS　The apoptotic cells induced by vincristine alone accounted for about 10% of all the cancer cells, while the percentage of apoptotic cells induced by a combination of vincristine with tetrandrine, neferine, or dauricine was found to be significantly higher than that by vincristine alone, and their reversal effects were positively correlated with the drug concentration and the exposure time. In addition, tetrandrine was shown to be the most potent in the reversal efficacy among the three compounds to be tested for apoptosis in vitro. CONCLUSION　Tetrandrine, neferine and dauricine showed obvious potenitiation of vincristine-induced apoptosis in the human mammary MCF-7 multidrug-resistant cells.

Objective To discuss the therapeutic effect in the treatment of the acute limb arterial critical ischemia.Methods Collect thirty-nine cases of acute limb arterial critical ischemia in Renji Hospital from Janary 2014 to July 2016.According to the patients' manifestation,these operations were porfermed including thrombectomy,cathetery-directed thrombolysis,mechanical suction bolt,percutaneous angioplasty and stenting.The effect and complications were observed.Results The eighteen patients in 39 cases (46.2％) were dead,including 5 cases without operation,13 operation.The eight cases were amputated during 34 cases of operations.In the 21 out-patients safely,2 cases were not followed up.The time of follow-up was from 3 to 27 months,on average 14.3 months.During the 21 patients,5 cases died from heart cerebrovascular or tumor diseases,3 cases with footdrop,2 cases with toe amputations,3 cases with distal leg and foot anesthesias.Conclusions The patiens with acute limb arteries critical ischemia must be treated as early as,and reinforced the management of multiple organ function,which may improve the diseases' therapeutic effect.

Objective To investigate the diagnostic value of the recombinant surface antigen 1 (rSAG1) in immunodiagnosis of toxoplasmosis. Methods Isopropyl β-D- 1 -thio-galaetopyranoside (IPTG) was used to induce the expression of recombinant plasmid pET28a-SAG1 of Escherich coli(pET28a-SAG1/BL21 ). The expression products (rSAG1) of pET28a-SAG1/BL21 were identified by Western blotting. The serum of mice infected with Toxoplasma gondii tachyzoites, normal mouse serum and the serum from 10 toxoplasma gondii patients were used as primary anti-Toxoplasma gondii antibodies, and the rSAG1 gene products were identified by Western blotting, by which the diagnostic value of rSAG1 in Toxoplasmosis was compared. Results After induction and purification, rSAG1 protein was obtained and its relative molecular mass was 38.5 × 103. The fusion protein could be recognized by the serum of mouse infected with Toxoplasma gondii tachyzoites, rSAG1 of expression products of surface membrane antigen SAG1 gene from Toxoplasma Gondii could be detected in 4 cases from 10 patients by Westem blotting.Conclusion The rSAG1 has a potential value in the immunodiagnosis of Toxoplasmosis.

Salvia miltiorrhiza is a highly valued traditional chinese medicine for the treatment of atherosclerosis-related disorders in china, such as cardiovascular and cerebrovascular diseases in China. The wilt disease is serious in the culture of S. miltiorrhiza. Wilt disease cause biomass of plant shoots and roots is lessened, active components are decreased. To solve these problems, we research the pathogen causing wilt disease of S. miltiorrhiza. The suspected pathogen is identified by morphology and etiological test. The identification was further confirmed by alignment the sequences of internal transcribed spacer (ITS) amplified by PCR. Our result show the wilt disease of S. miltiorrhiza mostly occurred in July and August, which is hot and wetter. The wilt disease rate of S. miltiorrhiza continuous cropping for one year in S. miltiorrhiz stubble is 10%, but the wilt disease rate of S. miltiorrhiza continuous cropping for three years in S. miltiorrhiz stubble is 60%-70%. The root rot of S. miltiorrhiz caused by the wilt disease, so the wilt disease was mistaken for the rot root in production. Morphological characteristics show the pathogen is Fusarium oxysporum. The sequence of ITS wes determined and found by BLAST shared 99% identity to that of F. oxysporum f. sp. cucumerinum. So it comes to the conclusion that the causing agent of wilt disease on S. miltiorrhiza belongs to F. oxysporum.
DNA, Intergenic ; genetics ; Fusarium ; genetics ; isolation & purification ; physiology ; Plant Diseases ; microbiology ; Polymerase Chain Reaction ; Salvia miltiorrhiza ; microbiology ; Seasons

Background Corneal neovascularization (CNV) is an important cause of visual impairment and graft rejection after allograft corneal transplantation in inflammatory corneal diseases. The mechanisms and therapy relating to CNV are intensely investigated at all times. Objective This study was to evaluate the effect of eicosapentaenoic acid (EPA) on CNV induced by alkali cauterization and its mechanism. Methods The animal models of corneal neovasculation were induced in the right eyes in 72 Sprayue-Dawley rats by putting a piece of 3 mmfilter paper with 1 mol/L NaOH at the center of the cornea for 30 seconds. The rats were then divided randomly into the 0.02 mg EPA treatment group (24 rats) ,0.03 mg EPA treatment group (24 rats) ,model group (24 rats) and normal group (6 rats). EPA of 0.04 ml with doses of 0.02 mg or 0. 03 mg or saline solution of 0. 04 ml was injected subconjunctivally in model rats and immediately after cauterization. The presence of CNV and corneal edema were observed daily by slit lamp biomicroscope. 1,4,7 and 14 days after operation, corneal histopathological examination was performed by hematoxylin and eosin staining. The vascular endothelial cells were stained with CD34 by immunohistochemistry,and the expression of IL-1α,IL-6 mRNA and the nuclear factor-κBp65 ( NF-κBp65 ) proteins was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. The use of animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by Hebei Province( version 1998 ). Results Under the slit lamp, CNV grew slowly from days 2-4 with obvious corneal edema and defect of epithelium. Larger CNV area and less edema were seen from days 7-10. Maximal vessel growth was observed 14 days after injury with thinner vessels in the model group. Histological examination showed that part of the corneal epithelium was damaged;serious corneal edema, more inflammatory cells and a lot of CNV in the stroma were presented in the model group. However, repairing of the corneal epithelium without CNV ,light corneal edema and less inflammatory cells were found in both the 0. 02 mg EPA and 0. 03 mg EPA treatment groups 7 days after alkali cauterization. The relative area of CNV in the 0. 02 mg EPA treatment group was ( 15.80±6.43 )% and ( 11.06±2. 14)% ,and that in the 0. 03 mg EPA treatment group was (16. 10±7.41 )% and (11.06±2. 51 )%, showing significant reduction in comparison with the model group [ (84. 74±7.77)% and (89.63±7.50) % ] 7 days and 14 days after operation ( P＜0. 05 ). Stronger expression of CD34 in the vascular endothelial cells of the cornea stroma was observed in the model group and an absence of CD34 was observed in the EPA-treated groups on the 7th day. RT-PCR revealed that the expression of IL-1α mRNA and IL-6 mRNA was lower in the EPA treatment groups than the model group ( P＜0. 05 ), and Western blot analysis showed that the expression of NF-κB/p65 in the corneas in the EPA treatment groups was significantly lower than that in the model group on the 4th day after operation (P＜0.05).Conclusion Topical application of EPA suppresses CNV induced by alkali burn possibly by inhibiting the expression of NF-κB,IL-1α and IL-6.

Objective To explore the value of wound blush in predicting patients' ulcer healing whom with critical limb ischemia after revascularization.Methods Retrospectively analyze the clinical data of 173 cases of critical limb ischemia with ischemic ulcers under thetreatment of endovascular therapy followed the concept of angiosome.According to the condition of wound blush after endovascular therapy,by compared the difference of limb salvage rate and ulcer healing time,and try to analyze the value of wound blush in predicting ulcer healing in patients.Results Included in the study with a total of 173 cases (173 limbs),group wound blush(+) 109 patients,group wound blush (-) 64 cases,the age,proportion of male patients,smoking history,diabetes,coronary heart disease,chronic renal insufficiency,pre and post operative ankle brachial index,were no statistical difference between the two groups.The ulcer healing time of group wound blush (+) was significantly shorter than that of group wound blush(-) (P ＜ 0.05).The rate of ulcer healing in group wound blush(+) was significantly higher than that in group wound blush(-) (P ＜ 0.05).In group wound blush(+),the cumulative rate of limb salvage was statisticallyhigher than group wound blush (-) (P ＜ 0.05).By logistic regression analysis,wound blush(-) (OR =4.5,P ＜ 0.05),IRc revascularization (OR =2.6,P ＜ 0.05) were independent risk factors of ulcer healing.Conclusions The resoult of wound blush(+) shows a good distal perfusion of foot.It can be used as a predictive factor for critical limb ischemia ischemic ulcer healing,and wound blush (-) was an independent risk factor for ulcer nonhealing.

A novel method was developed for the rapid determination of multi-indicators in corni fructus by means of near infrared (NIR) spectroscopy. Particle swarm optimization (PSO) based least squares support vector machine was investigated to increase the levels of quality control. The calibration models of moisture, extractum, morroniside and loganin were established using the PSO-LS-SVM algorithm. The performance of PSO-LS-SVM models was compared with partial least squares regression (PLSR) and back propagation artificial neural network (BP-ANN). The calibration and validation results of PSO-LS-SVM were superior to both PLS and BP-ANN. For PSO-LS-SVM models, the correlation coefficients (r) of calibrations were all above 0.942. The optimal prediction results were also achieved by PSO-LS-SVM models with the RMSEP (root mean square error of prediction) and RSEP (relative standard errors of prediction) less than 1.176 and 15.5% respectively. The results suggest that PSO-LS-SVM algorithm has a good model performance and high prediction accuracy. NIR has a potential value for rapid determination of multi-indicators in Corni Fructus.
Algorithms ; Calibration ; Cornus ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Least-Squares Analysis ; Models, Theoretical ; Neural Networks (Computer) ; Quality Control ; Spectroscopy, Near-Infrared ; Support Vector Machine

The xylanolytic enzymes found in Thermotoga maritima showed extremely high thermostability and considerable potential in industrial application. Yet expression level of the genes encoding these enzymes was very low. The alpha-glucuronidase gene aguA from T. maritima ATCC 43589 was cloned and expressed in several E. coli strains with different vector. The alpha-glucuronidase was overexpressed in E. coli BL21-CodonPlus(DE3)-RIL with plasmid pET-28a(+), and made up about 20% of the total proteins present in the intracellular soluble fraction. The results proved the assumption that rare codons for arginine (AGA/AGG) and isoleucine (AUA) affect the expression of aguA gene from hyperthermophilic bacterium T. maritima in E. coli. Purification procedure included two steps, heat treatment and immobilized metal affinity chromatography, and over 13.5mg of pure enzyme was obrained from 1L of induced culture. The purified enzyme showed a single band on SDS polyacrylamide gel electrophoresis with a purification of 5.1 fold, and a yield of 55.1%. The optimum activity of recombinant alpha-glucuronidase was found at pH 6.0 and 85 degrees C, the enzyme retained 70% of its activity after 1 h of incubation at 85 degrees C. The induction conditions for expression of recombinant strain BL21-CodonPlus(DE3)-RIL/pET-28a-aguA were studied on induction time and duration by IPTG. The results showed that the activity of thermostable alpha-glucuronidase reach the maximum in 5-hour after inducted at the exponential phase (OD600 of 0.7 - 0.8).
Escherichia coli ; genetics ; Glycoside Hydrolases ; genetics ; isolation & purification ; metabolism ; Plasmids ; Recombinant Proteins ; biosynthesis ; isolation & purification ; Thermotoga maritima ; enzymology

OBJECTIVETo investigate the molecular mechanism of haemophilia B caused by the novel mutation of Arg327Ile (R327I) in FIX gene.

METHODSThe R327I, R327Ala(A), R327Lys(K), R327Asn(N) and a replacement mutant (FIXβFVII), in which FIX β strand 324-329 was replaced by that of FVII 298-303, expression plasmids were constructed with site-directed mutagenesis method based on the wild-type (WT) FIX expression plasmid. The HEK293 cell was transiently transfected, then the activity of FIX (FIX:C) was assayed by one stage method in the conditioned medium, while the FIX:Ag in both the conditioned media and the cell lysates was measured by ELISA. The molecular weight and the semi-quantity of expressed FIX were analyzed by Western blot. Fluorescent protein expression plasmid was constructed to investigate the synthesis and secretion of the FIX R327I mutation in the viable cells.

RESULTSFIX:C of the R327I mutant protein was 4.49% of the level of the WT in the conditioned medium, and the FIX:Ag of the R327I mutant protein in the conditioned medium and the cell lysates was 31.02% and 129.29% compared to that of WT, respectively. The mutation was characterized as cross-reaction material reduced (CRMR). The viable cell fluorescent assays showed that the R327I protein was more in both the viable cells and in lysosome than that of WT. The FIX:C of the R327A, R327K, R327N and FIXβFVII mutants was reduced compared to that of WT, the reduction of FIX:C of FIXβFVII was the most significantly amount among all the mutants in medium. FIX:Ag of all the mutants in the medium, except that the R327K increased, was reduced. The result of Western blot showed that the molecular weight of R327I protein was the same as that of WT, but the amount of the protein was much less compared with WT in the conditioned medium.

CONCLUSIONThe abnormal synthesis and secretion as well as the abnormal function of the R327I mutant protein causes haemophilia B. The residue of R327 as well as the β strand domain of R327 located play important roles of the specific function of FIX.

Factor IX ; genetics ; HEK293 Cells ; Hemophilia B ; genetics ; pathology ; Humans ; Mutagenesis, Site-Directed ; Mutation ; Transfection