Objective To evaluate free or pedicled perforator flaps for repairing chronic osteemylitis with soft-tissue defect in the distal lower extremity. Methods From May of 2006 to October of 2007, 28 consecutive patients of chronic osteomylitis with soft-tissue defect in the distal lower extremity underwent surgical debridement and reconstruction with free or pedicled perforator flaps. There were 13 free flaps. The free anterolateral thigh flaps were used in 2 cases to repair the soft defects in the front of leg, 3 cases in the front of the malleolus, 2 cases in the dorsum of foot, 2 cases in the heel. The free lateral crural flaps nourished by perone al artery were used in 4 cases to repair the soft defects in the dorsum of foot. There were 15 pedicled flaps. Posterior tibial artery perforator flaps were used in 4 cases to repair the soft defects in the front of leg, and 2 cases in the medial malleolus. Lateral retromalleolar perforator flaps nourished by peroneal artery were used in 6 cases to repair the soft-defects in the heel, 1 case in the lateral malleolus and 1 case in the dorsum of foot, the first dorsal metatarsal artery perforator flap was used to repair the proximal dorsum of hallux. The wound was closed with irrigation-suction in 7 cases and with vancomycin-impregnated gelatin in 8 cases. Results All 27 flaps were successfully survived except insuffcient vein refluence in 1 posterior tibial artery perforator flap, which resulted in a superficial necrosis and healed spontaneously. The follow-up period from 6 months to 2 years revealed that recurrence developed in two diffuse type patients and both were treated once and twice with success, respectively. The others healed without any signs of recurrences. No debulking procedure was necessary in any case. Secondary bone graft was performed in 3 cases. All patients were ambulatory and fully weight-bearing with normal clinical parameters at the time of last review. According to the evaluating criteria for the treatment of foot disease, the mean score was 84.5. Conclusion Free or pedicled perforator flap has been shown to be well vascularised, and it is feasible for the treatment of chronic osteomyelitis with soft-tissue defect in the distal lower extremity.

Objective During pregnancy , exosomes can be released from the placenta into maternal circulation and play im-portant roles in normal pregnancy or placenta-related diseases .We aimed to establish a simple and efficient method for isolating and i-dentifying placental exosomes from maternal serum and lay a foundation for the studies of pregnancy -related diseases . Methods Using sucrose gradient centrifugation with 8% PEG6000 precipitation twice , we isolated and purified placenta-derived exosomes from normal maternal serum and detected their molecular markers CD 63 , CD81 and PLAP by Western blot , followed by silver staining anal-ysis of the protein profile of the exosome pellet .We identified the morphology of the placenta-derived exosomes by transmission electron microscopy ( TEM) and measured the size and distribution of the particles by dynamic light scattering ( DLS) . Results Silver stai-ning of the protein profiles of the exosomes after sucrose gradient centrifugation clearly revealed the bands of the protein molecules . Western blot showed the expressions of CD 63, CD81, and PLAP in the 21-34%density layer, which demonstrated the presence of serum placental exosomes mainly in the 1.09-1.16 g/mL density layer.TEM exhibited that the placenta-derived exosomes were round or oval cup-shaped, specifically expressing PLAP, and the particles were uniform in size, with a mean diameter of (41.79 ±11.94) nm. Conclusion A simple, fast, and efficient method was successfully established for isolating placenta-derived exosomes from ma-ternal serum, which provides a basis for studying the roles of placental exosomes in normal pregnancy and placenta -related diseases.

OBJECTIVE To study the molecular mechanism of cisplatin(DDP)by which HeLa cell growth and proliferation are inhibited. METHODS Cultured HeLa cells were treated with DDP 0.02-75 μmol · L-1 for 24 or 48 h. CCK-8 assay was used to determine the cell proliferation. The wound scratch assay was used to detect the cell migration and invasion. Flow cytometry was used to detect the cell cycle arresting. q-PCR was used to test the expression of metastasis suppressor gene 1 (MTSS1)mRNA. Western blot was used to determine protein levels of MTSS1,phosphorylated-extra?cellular signal-regulated kinase(p-ERK) and phosphorylated-serine-threonine kinase(p-AKT). RESULTS Following the treatment with DDP for 24 or 48 h,the proliferation of HeLa cells was inhibited significantly (P<0.05),the value of the half inhibitory concentration (IC50) of cells was 4.14 and 11.82 μmol · L-1. Migration and invasion activity of HeLa cells were reduced according to the wound scratch assay(P<0.05). Flow cytometry results showed that the cell cycle was arrested at S phase. q-PCR results showed that MTSS1 mRNA expression changed with DDP in a concentration-dependent manner (r24 h=-0.965,P<0.01;r48 h=-0.953,P<0.01). Western blot showed that the protein levels of MTSS1,p-ERK and p-AKT expression declined significantly with the increase in DDP concentrations(p-ERK:r24 h=-0.875,P<0.01;r48 h=-0.966,P<0.01. p-AKT:r24 h=-0.831,P<0.01;r48 h=-0.863,P<0.01. MTSS1:r24 h=-0.969,P<0.01;r48 h=-0.988,P<0.01). CONCLUSION DDP treatment inhibits HeLa growth and proliferation by interfering with the MTSS1 expression and disturbing the activation of ERK and AKT signaling pathways.

Objective To investigate the effects of cyclopamine (CYP) on endometrial carcinoma (HEC-1A) cell survival and on induction of cell apoptosis .Methods HEC-1A cells were treated with various doses of CYP (0, 5,10, 20 and 40 μmol/L) for 24 h respectively .Then,the inverted microscope was used to observe cell morphology .Cell proliferation and apoptosis were tested by CCK-8 assay and AO/EB bi-labelling assay.The apoptosis rate of HEC-1A was analyzed using flow cytometric analysis , and the key gene expression of Bax and Bcl-2 was detected by quantitative PCR .Results The HEC-1A cells exhibited dramatic morphological changes after treatment with CYP and in a dose-dependent manner .CYP significantly inhibited HEC-1A cell proliferation using CCK8 assays(P<0.05), and induced cell death by AO/EB bi-labelling assay.Moreover,flow cytometry analysis showed that CYP treatment resulted in HEC-1A cell apoptosis, and that a higher concentration of CYP induced severer cell apoptosis (P<0.05).Meanwhile, CYP treated HEC-1A cells exhibited up-regulated expression of Bax and down-regulated expression of Bcl-2 according to Q-PCR.Conclusion Our findings indicatee that CYP can inhibit HEC-1A cell proliferation and induce cell apoptosis .

Objective To investigate the application and clinical outcomes of anterolateral thigh perforator flap bridging different superficial circumflex iliac artery flaps for the treatment of complex limb wound.Methods Data of 10 patients,who had undergone free tissue transfer of anterolateral thigh perforator flap bridging different superficial circumflex iliac artery flaps (iliac osteocutaneous flap,iliac crest flap and ilioinguinal flap) for the complex limb wound from March 2009 to January 2011,were retrospectively analyzed.There were 9 males and 1 female,aged from 21 to 57 years (average,39.7 years).Iliac osteocutaneous flap was used in 7 patients,iliac crest flap in 2 patients and ilioinguinal flap in 1 patient.The free tissue transfer was performed for hand reconstruction in 3 cases,foot reconstruction in 4 cases and leg reconstruction in 3 cases.The mean size of soft tissue defect was 20 cm × 9.7 cm.Results The mean size of the anterolateral thigh flap and the ilioinguinal flap was 17.8 cm×9.4 cm and 8.4 cm×4.5 cm,respectively.The mean volume of iliac crest was 5.4 cm×2.1 cm×0.8 cm,and the mean length of flow-through conduit was 10.5 cm.Except for the distal necrosis of iliac osteocutaneous flap in 1 case,which were treated by dressing change and skin grafting,the other flaps survived without complications.All patients were followed up for 6 to 36 months (average,12 months).The average bone union time was 3 months in the hand group,4 months in the foot group,and 4.5 months in the leg group.The disabilities of the arm,shoulder,and hand questionnaire score averaged 43,the mean Japanese Orthopaedic Association's foot rating scale was 71.3,and the mean Puno's functional score was 91.Donor site scars were mostly flat,pale and soft but widened in the thigh in 6 patients and in the ilioinguinal region in 3 patients,and numbness was reported in 2 patients.Conclusion In clinical situation that requires for coverage of complex and extensive limb defects,the free tissue transfer of anterolateral thigh perforator flap bridging superficial circumflex iliac artery flap is a better method,which can meet specific reconstructive demands with minimal donor site morbidity.However,a relative high risk has to be considered.

Nanobody is a kind of antibody from camel, which misses light chain. Nanobody has the same antigen binding specificity and affinity as mAb. Moreover, because of its small molecular weight, high stability and easy preparation, nanobody has great value of biomedical applications. In this study, we successfully prepared highly pure antiEGFR nanobody in E.coli using genetic engineering techniques. Cell proliferation assay (CCK-8 assay) and migration experiments (cell scratch test and Transwell assay) indicated that the recombinant antiEGFRnano can significantly inhibit the proliferation and migration of endometrial cancer cells. These results provide a new way of thinking and methods for EGFR-targeted therapy of endometrial cancer.

Objective To explore the correlation of nurses' quality of life with sleep quality and their job burnout in class Ⅲ grade A hospitals, and to identify the high risk factors affecting nurses' quality of life, so as to provide constructive suggestions.Methods From September 2015 to January 2016, totally 460 nurses working in class Ⅲ grade A hospitals of Xuzhou were investigated by self-administered questionnaire using the Pittsburgh Sleep Quality Index (PSQI), the Maslach Burnout Inventory (MBI) and the World Health Organization quality of life instrument's BREF (WHOQOL-BREF). The data was analyzed by SPSS17.0 statistical software, and measurement data uset-test. The multivariate linear regression equation model took PSQI and MBI entries as independent variables, and took the dimensions of WHOQOL-BREF as the dependent variable, and used the stepwise method to run the equation.Results A total of 460 questionnaires were issued and 400 valid were recovered. The effective recovery rate was 86.9％. Compared with respective norm, the scores of PSQI (7.18±3.47), emotional exhaustion (23.99±12.45) and depersonalization (8.20±5.97) dimension of MBI were higher, and the differences were statistically significant (t=38.846, 36.154, 25.737;P<0.001). The score of personal accomplishment dimension of MBI and all four areas in WHOQOL-BREF were lower than norm, and the differences were statistically significant (P<0.001). Regression analysis showed that subjective sleep quality, sleep time in PSQI and emotional exhaustion area, personal accomplishment area of MBI had significant effects on nurses' quality of life (P<0.05).Conclusions The quality of sleep and quality of life of clinical nurses in class Ⅲ grade A hospitals is poor, and their job burnout is strong. It suggests that solving nurses' working life dilemma should start from the high risk factors which have great influence on it, so as to improve nurses' quality of life.

OBJECTIVE: To explore the genetic basis for a pedigree affected with X-linked recessive mental retardation Claes-Jensen type. METHODS: Genomic DNA was extracted from peripheral blood samples of the patient, his parents (phenotypically normal) and two elder brothers with similar clinical manifestations. Whole exome sequencing was carried out for the proband, and the result was verified by Sanger sequencing. RESULTS: The proband was found to harbor a hemizygous c.1565C>T missense variant in exon 11 of the KDM5C gene. The transition has resulted in replacement of serine by phenylalanine at position 522 (p.Ser522Phe). Sanger sequencing showed that the patient's two elder brothers and mother carried the same variant, which was predicted to be probably damaging by SIFT, PolyPhen2 and Mutation_Taster. The three affected brothers presented with similar clinical phenotypes characterized by mental retardation, speech delay, behavioral problem, self-limited epilepsy responsible to medication, short stature and microcephaly. The mother only had mild cognitive impairment and learning disability. The same variant was not found in their father and was unreported previously. CONCLUSION The c.1565C>T (p.Ser522Phe) of the KDM5C gene probably underlay the X-linked recessive mental retardation Claes-Jensen type in this pedigree.
Aged ; Female ; Histone Demethylases/genetics* ; Humans ; Male ; Mental Retardation, X-Linked/pathology* ; Mutation, Missense/genetics* ; Pedigree ; Phenotype ; Whole Exome Sequencing

BACKGROUND:Gastrodin has anti-inflammatory effects and is mainly used in clinical practice for the treatment of ischemic stroke,and its mechanism of action is still unclear. OBJECTIVE:To explore the mechanism of gastrodin intervention on inflammatory injury in ischemic stroke rats. METHODS:Fifty Sprague-Dawley rats were divided into sham-operated group,model group,positive control group,high-dose gastrodin group and low-dose gastrodin group by the randomized numerical method,with 10 rats in each group.Ischemic stroke models were established by the middle cerebral artery occlusion method in all groups of rats except for the sham operation group.Administration in each group started on the 3rd day after surgery,and the rats in the positive control group were intraperitoneally injected with edaravone injection(6 mg/kg),the rats in the high-and low-dose gastrodin groups were intraperitoneally injected with 50 and 10 mg/kg gastrodin injection respectively,and the rats in the sham-operated and model groups were intraperitoneally injected with the equal volume of physiological saline.After 14 days of continuous treatment in each group,the pathological changes in rat brain tissue were observed,and the positive expression of NLRP3 inflammasome and the expression of inflammatory response-related proteins and their mRNAs were detected. RESULTS AND CONCLUSION:Compared with the sham-operated group,the volume of cerebral infarction became larger in the model group;the structure of brain tissue was loose,irregular cavities could be observed,and the number of neurons was reduced and irregularly arranged;the positive expression of NLRP3 inflammasome increased(P<0.01);and the protein and mRNA expression levels of Toll-like receptor 4,myeloid differentiation factor 88,apoptosis-associated speck-like protein containing a caspase-recruitment domain,Caspase-1,and interleukin-1β increased(P<0.01).Compared with the model group,the volume of cerebral infarction became smaller in the high-and low-dose gastrodin groups;the neurons were regularly arranged,increased in number,and uniformly distributed;the positive expression of NLRP3 inflammasome was decreased(P<0.05);the protein and mRNA expression levels of Toll-like receptor 4,myeloid differentiation factor 88,apoptosis-associated speck-like protein containing a caspase-recruitment domain,Caspase-1,and interleukin-1β were decreased in the high-dose gastrodin group(P<0.01);Toll-like receptor 4 protein expression showed no significant changes in the low-dose gastrodin group,and the protein and mRNA expression of the other inflammatory response-associated factors decreased(P<0.05,P<0.01).To conclude,gastrodin attenuates inflammatory injury in ischemic stroke rats,and its mechanism of action may be related to the inhibition of inflammatory response-associate factor expression.

Temporal bone CT imaging is particularly important for diagnosis and treatment of ear diseases,but there are some issues such as insufficient resolution,strong artifacts and low signal-to-noise ratio under low radiation doses,which limit its'clinical application.Recent years,deep learning(DL)had shown great potential in classifying,segmenting and reconstructing medical images,providing new ideas for improving imaging quality of temporal bone CT.The application progresses of DL in temporal bone CT imaging were reviewed in this article.