Objective: To develop a quantitative PCR method for detecting hookworm infection and quantification. Methods:A real-time PCR method was designed based on the intergenic region II of ribosomal DNA of the hookworm Necator americanus. The detection limit of this method was compared with the microscopy-based Kato-Katz method. The real-time PCR method was used to conduct an epidemiological survey of hookworm infection in southern Fujian Province of China. Results:The real-time PCR method was specific for detecting Necator americanus infection, and was more sensitive than conventional PCR or microscopy-based method. A preliminary survey for hookworm infection in villages of Fujian Province confirmed the high prevalence of hookworm infections in the resident populations. In addition, the infection rate in women was significantly higher than that of in men. Conclusions: A real-time PCR method is designed, which has increased detection sensitivity for more accurate epidemiological studies of hookworm infections, especially when intensity of the infection needs to be considered.

Objective: To establish and authenticate the chloasma mouse model with syndrome of liver-qi stagnation,and compare it with other existent models.Method: After the injection of progesterone,chronically constrained them and gave them partially ultraviolet radiation.Results: Multiple models of progesterone injection and ultraviolet radiation compounding with chronic constraint can heighten MDA,depress SOD,and increase melanocytes of skin.Meanwhile,the ethology of this model showed it's syndrome of liver-qi stagnation.Conclusion: The multiple factor chloasma mouse model of liver-qi stagnation syndrome has been built successfully.

Objective To investigate the effect of modified suture on the maternal skin healing, scar healing and satisfaction to cesarean section. Methods A total of 244 cases of cesarean section in our hospital from January 2015 to January 2016 were randomly collected and divided into observation group (n=143) and control group (n=101). Patients in the observation group were treated with the modified suture method, and the control group was given the conventional method to suture the skin. The VAS score, suture time and complication rate were observed in the 2 groups. Results The VAS scores were significantly lower at one week and one month after operation (1.15±0.43 and 0.07±0.01, respectively) than those of the control group (1.40±0.51 and 0.13±0.03, respectively, P<0.05). The suture time were significantly shorter in the observation group (5.12±0.95) than those of control group (7.28±0.83, P<0.05). Postoperative complication rate was significantly lower in observation group (2.80%) than that of control group (9.90%, P<0.05). The comfort level to incision and satisfaction rate to scar were significantly higher (94.41% and 95.10%, respectively) than those of control group (83.17% and 87.13%, respectively). The average thickness of scars at day 90 and day 180 after operation were significantly less in the observation group [(2.14±0.32) mm and (3.08±1.02) mm, respectively] than those of the control group [(3.10±0.48) mm and (5.22±1.10) mm, respectively, P<0.05)]. Conclusion The improved suture method after cesarean section has the advantages of shorter suture time, less poor wound healing, higher maternal satisfaction rate.

Objective To study the healing response of a tissue engineered bone of bMSCs combined rFN/CDH-BCP in a rabbit model. Methods The biomimetic surface was achieved by immobilizing rFN/CDH onto biphasic calcium phosphate ceramic ( BCP) covalently. The effect of rFN/CDH-BCP on adhesion,proliferation and differentiation were evaluated comprehensively by using cell centrifugal adhesive as-say,MTT method,SEM,Alkaline phosphatase ( ALP) activity and alizarin red staining respectively. A rabbit lumbar fusion model was estab-lished by implanting bMSCs combined rFN/CDH-BCP into intertransverse process space of L4 ~L5 ,the fusion site bone formation was ob-served by imaging means,histological techniques was used to observe the new bone formation and distribution of seed cells. Results Cell centrifugal adhesive assay indicated the adherent bMSCs on rFN/CDH-BCP associated with the ligand density,the biomimetic surface posses-ses excellent biocompatibility. The ALP activity on rFN/CDH-BCP surface was the highest among all samples on the 10th day after induction (P<0. 05). On the 21st day,alizarin red staining showed that the oval-shaped and orange-red nodules,either the number or the area,distrib-uted wider on rFN/CDH-BCP surface. The results from X plain after 3 months revealed a fuzzy gap between material and bone bed,and higher cover rate of intertransverse process space with new bone deposition on rFN/CDH-BCP surface. Conclusion Histologically,rFN/CDH-BCP exhibited as interlacing bone trabecula bridging biomaterial suface and cortical bone of transverse process continously. The bone mass was much more than the ture BCP class. The comprehensive data reveals that when loaded with MSCs,rFN/CDH-BCP demonstrates superior char-acteristics of osteoconduction and osteoinduction,and substantially enhances healing capacity in vivo.

The main aim of this study was to transform the enhanced green fluorescent protein gene (egfp) into biocontrol fungus Paecilomyces lilacinus strain 9410. We constructed the expression vector pUPNGT of the fusion gene nptII-egfp using pcDNA3.1(-) as a helper plasmid. The egfp gene was then transformed into P. lilacinus strain 9410 via Agrobacterium tumefaciens-mediated transformation. PCR and Southern blotting analysis showed that the egfp gene was integrated into the genomes of the tested transformants and the integration manner was single-copy. The transformants could generate green fluorescence when they were excited by 488 nm blue laser. These results indicated that the egfp gene had been successfully transformed into P. lilacinus 9410 and expressed in the tested transformants. Our work may provide a new approach to assess environmental safety and practical biocontrol efficacy ofP. lilacinus under different conditions.
Agrobacterium tumefaciens ; genetics ; Green Fluorescent Proteins ; genetics ; Paecilomyces ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Transformation, Genetic

Osteoporosis is a progressive metabolic disease. In traditional Chinese medicine (TCM), deficiency of the liver, spleen and kidney and blood stasis can induce osteoporosis. The main etiology was kidney deficiency which harmed the liver and spleen and causeqi-blood deficiency and blood stasis. The insufficiency of natural endowment and postnatal malnutrition caused poor nutrition of tendons and vessels, which induced debility of bone. This article discussed the etiology and pathogenesis of TCM understanding on osteoporosis in details. Opportunities and challenges of TCM in osteoporosis treatment were explored. The homology of Chinese medicine and food as well as the long-term of osteoporosis showed that TCM had incomparable superiority in osteoporosis treatment. While, the scientific research methods and reasonable evaluation of TCM safety were important to display its advantages in osteoporosis treatment.

Objective To discuss surgical method and clinical curative effect of the headless compression cannulated screw percutaneous minimally invasive treatment of Evans -JensenⅠ,type Ⅱ of intertrochanteric fracture. Methods The clinical data of 21 cases of Jensen Evans and type II femoral intertrochanteric fractures who were treated by headless compression cannulated screw minimally invasive treatment were retrospectively annalyzed.In all patients,male in 6 cases,female in 13 cases;63 -86 years old,average 71.79 years.According to the Evans -Jensen norm and curative effect evaluation according to the improved Harris standard.Results Follow -up time was 6 -18 months (mean 12.15 months),two of them respectively in the six months after death from acute myocardial infarc-tion (ami)and 11 months of cor pulmonale,the rest of 17 cases of bony healing.Harris,the average score (67 -95points)at the time of the last follow -up,including 14 cases of optimal good in 2 cases,fair 1,poor in 1 case,was 88.89%.Conclusion Headless compression cannulated screw percutaneous minimally invasive treatment of Evans-Jensen Ⅰ,type Ⅱ intertrochanteric fractures has the advantangtages of minimally invasive,reliable fixation,high rate of fracture healing,allowing early hip not functional exercise load conditions and obtaining ideal therapeutic effect.

AIM:To investigate the changes of brain-derived neruotrophic factor(BDNF),tyrosine kinase B(TrkB) in rat cortex and hippocampus with chronic immobilization stress and the influence of three Chinese formulas(Xiaoyaosan,Sijunzitang,Jinkuishenqiwan) on them.METHODS:Chronic immobilization stress method(180 min daily,repeated 7 days or 21 days) was taken,and the changes of BDNF,TrkB in rat forehead cortex and hippocampus CA1 were measured by immunohistochemistry integrated image analysis.RESULTS:The contents of BDNF in rat forehead cortex and hippocampus CA1 were obviously lower in the model group of 7 days and 21 days than those in the normal control group(P

Objective To investigate the safety and efficacy of three column osteotomy (3-CO) procedures through previous spinal fusion site for the revision surgical treatment in severe spinal deformity patients.Methods From Oct.2010 to May 2014 in our hospital,a total of 12 severe spinal deformity patients underwent 3-CO for the revision surgical treatment.There were 7 males and 5 females with the average age of (21.8±3.8) years,ranging from 18 to 30 years.The mean time from the initial operation to the revision surgery was (10.2±4.8) years (ranging from 3 to 17 years).The reasons for revision were:curve progression in 7 patients,neurologic deficit in 2 cases,implant failure in 1 patient and pseudarthrosis in 2 patients.The coronal parameters including major Cobb angle and distance between C7 plumb line and center sacral vertical line (C7PL-CSVL),and the sagittal parameters including global kyphosis curve and sagittal vertical axis (SVA) were measured pre-operatively,post-operatively and at last followup,respectively.The operation time,intraoperative blood loss and complications were recorded.The paired t test was used to evaluate the difference among pre-revision,post-revision and last follow-up.Results The average operation time was (451.7±83.1) min (range,320-600 min) and the average blood loss was (4 016.7± 1 080.0) ml (range,2 700-6 000 ml).The average follow-up time after revision operation was (35.4±9.8) months (range,24-49 months).The coronal Cobb angles of pre-revision and post-revision were 83.8°±23.3°and 34.6°± 13.7°.The average correction rate was 60.1％ ±8.8％.At last follow-up,the average coronal Cobb angle was 34.9°±13.8°,there was no significant loss of correction.The pre-revision and post-revision values of global kyphosis were 99.1°±13.3°and 38.7°±7.8° with a mean correction rate of 60.8％ ±6.7％.At the last follow-up,the average global kyphosis was 39.3°±7.5°and no loss of correction was found.For the C7PL-CSVL and SVA,pre-revision (30.3± 17.1) mm and (40.1±31.1) mm were corrected to (14.3 ±7.6) mm and (19.1± 12.3) mm immediately after revision operation,respectively.At final follow-up,the average C7PL-CSVL and SVA were(14.1 ± 7.6) mm and (19.6± 12.1) mm,the correction was well maintained.Obviously,two patient's neurological status improved from Frankel C before revision surgery to Frankel E.Complications were encountered in five patients (41.7％),including SEP signal changed in 1 patient (8.3％),transient neurologic deficit after revision surgery in 1 patient (8.3％),cerebrospinal fluid leak in 1 patient (8.3％),and pleural effusion in 2 patients (16.7％).During the follow-up time,there was no patient experienced pseudarthrosis,implant failure,infection or significant loss of correction.Conclusion Based on results of this study,it was concluded that 3-CO procedures through previous spinal fusion sites could obtain satisfactory and safety results in severe spinal deformity revision surgery.However,it is a technique-demanding procedure with more blood loss,longer operative time and higher risk of perioperative complications.

Depression and anxiety is one of common mental illness that are being seriously harm to the physical and mental health of human being in recent years.Depression and anxiety are difficult to be well treated with a single and targeted drug,owing to its biological-psychological-social interaction pathogenic feature.Due to its own multicomponent characteristics,traditional Chinese medicine (TCM) compound Xiao-Yao (XY) powder can exert the antidepression and anxiety effects from multi-level,multi-link and multi-target.Based on previous studies on effect and mechanism of XY powder on anti-depression and anxiety of our group,we proposed that XY powder had two main characteristics when treating depression and anxiety,which were "two-way adjustment" and "multi-target,multisystem,multi-level".Additionally,to further clarify the pharmacodynamic material basis of XY powder,more importance is that we should depend on the variety of disciplinary research methods,focused on the TCM syndrome and the combination of disease and TCM syndrome,brought both disease evaluation and syndrome evaluation method,took the importance of gut-brain axis (GBA),and combined with the correlation between the formula and the syndrome.