BACKGROUND: Human allogenic acellular dermal matrix, as a kind of permanent dermal scaffold, has widely used in the fields of burn wound reparation and aesthetic medicine. However, it is limited due to insufficient resources. OBJECTIVE: To prepare porcine acellular dermal matrix (PADM) dermal matrix, in addition, to estimate its in vitro biocompatibility. DESIGN, TIME AND SETTING: An in vitro cytology contrast experiment. The Experiment was performed at the laboratory of Biomaterials and Pharmacy Technology Institute, China Institute for Radiation Protection between August 2007 and June 2008. MATERIALS: The experiment pigs were supplied by experimental animal center of China Institute for Radiation Protection. Human fibroblasts were obtained from prepuce tissues of healthy children who underwent circumcision at the Shanxi Provincial General Hospital, Chinese People's Armed Police Forces. METHODS: The PADM was prepared from porcine skin by removing epidermis with a hypertonic salt solution and excluding cellular components in dermis with sodium dodecyl sulfate, and trypsin in hypersonic conditions. Human fibroblasts were cultured in vitro, and the biocompatibility of PADM was estimated with MTT method and contact method. MAIN OUTCOME MEASURES: ①Histological morphology of PADM. ②In vitro biocompatibility of PADM. RESULTS: The prepared PADM was a kind of matrix with normal structure and organization of collagen without epidermis and any cellular components in the dermis. The extraction of the porcine acellular dermal matrix had no effect on proliferation of the cultured human fibroblast. The cultured human fibroblasts could attach and proliferate on PADM. CONCLUSION: The PADM effectively removed epidermis and all cellular components with excellent biocompatibility can be obtained by this preparation method.

Objective To evaluate the expression of human vascular endothelial cell growth factors 165 (hVEGF165) gene transfected into fibroblasts by recombinant adenovirus and study the repairing effect of this cells on radiated skin ulcer in rats.Methods The recombinant adenovirus with hVEGF165 was established and transfected to rat primary fibroblasts, and its expression of hVEGF165 in fibroblasts was identified with real-time PCR, immunocytochemistry and Western blot.Twenty four clean grade SD rats of were irradiated locally with 50 Gy γ rays to generate an animal model of radiation skin injury.The hVEGF165-transfected cells were injected to the irradiated site under rat skin 7 d post-irradiation.The therapeutic effects on the irradiated skin wound were evaluated through general observation as well as histological staining of HE.The expression of hVEGF in the irradiated skin tissue with fibroblasts injection was analyzed by Real-time PCR.Results The hVEGF165 gene was overexpressed in the transfected cells and approached to 88 373-fold bigger compared to controls transfected with blank vectors, and an extensive expression of VEGF in the cytoplasm of transfected cells was observed by immunohistochemistry.VEGF protein with the relative molecular mass of 23 000 was also detected in cell lysate by Western blot.The local skin ulcers in rats occurred about two weeks after irradiation.In the hVEGF165-transfected group, the average area of radiation-injured skin was 40.2 mm2, about 57％ less than that of the control group transfected with blank vectors so that the healing time was shorten by 6 days.The relative concentration of hVEGF mRNA in the skin tissue of rats injected with hVEGF165-transfected cells were 5.15-fold and 4.15-fold bigger compared to that of controls (t =3.385,3.220, P ＜ 0.05) at 3 and 7 d after administration.Conclusions The primary fibroblasts transfected with hVEGF165 gene could efficiently release VEGF to the irradiated skin tissue and promote the recovery of irradiation skin ulcers by shortening the healing time and thus enhanced the therapeutic effect on skin wounds.

AIM:To explore the effect of HbA1c level in the patients with type 2 diabetes mellitus (T2DM) on the rheological properties of erythrocytes and the structure of hemoglobin (Hb).METHODS:The patients with T2DM were classified into 3 groups:the patients with good glycaemic control (group A, HbA1c<7.0%), the patients with poor glycaemic control (group B, 7.0%≤HbA1c<9.0%) and the patients with persistent hyperglycemia (group C, HbA1c≥9.0%) .The rheological properties of a single living erythrocyte were analyzed by the techniques of static imaging and anal -ysis.Ultraviolet-visible spectroscopy was used to study the structure of Hb .RESULTS: Compared with group H , the roundness effect factor (REF) of erythrocytes, which was positively related with HbA1c level, was significantly increased in groups A, B and C (P<0.05).The contact area of erythrocytes in group B and all the morphological parameters of erythrocytes in group C were significantly higher than those in group H ( P <0.05 ) .There were significant differences among groups A , B and C in deformation capacity and elastic parameters of the cell membrane ( P<0.05 ) , but with no difference in the long axis and short axis between group A and group B .Compared with group H , no obvious change in the spectral pattern and spectrum peak of Hb in groups A , B and C was observed.However, the absorbance of Hb, which showed a trend of gradual decline with the increase in HbA 1c level, in group B and group C was significantly decreased as compared with group H (P<0.01), and the Hb absorbance in group C were also lower than that in group A (P<0.05). CONCLUSION:With the increase in HbA1c level, the morphology along with the deformation function of erythrocytes in T2DM changes and declines gradually , and the structure of Hb may also change .

BACKGROUND:It is important to prepare a mal eable putty to repair irregular bone defects because the preformed bone substitute material cannot suit to bone defects.
OBJECTIVE:To prepare mal eable demineralized bone matrix/hyaluronic acid putty and to screen out the optimal composite ratio, as wel as to evaluate its cellcompatibility.
METHODS:Demineralized bone matrix was prepared from the cortical bone of healthy donors. The hyaluronic acid was dissolved at concentrations of 0.75%, 1.5%, 3%, 4.5%, and the viscosity of them was measured. Then 450 mg demineralized bone matrix was compounded with 1 mL hyaluronic acid solution to prepare the demineralized bone matrix/hyaluronic acid putty, and the optimal ratio of demineralized bone matrix/hyaluronic acid was screened by the disintegrated time of the putty at 37 ℃. L-929 mouse fibroblasts were cultured in the leaching liquor of demineralized bone matrix/hyaluronic acid, and the cytotoxicity was tested by cellcount kit-8 assay at days 1, 4 and 7.
RESULTS AND CONCLUSION:With the increase of hyaluronic acid concentration, the viscosity of hyaluronic acid solution was increased;the demineralized bone matrix/hyaluronic acid putty was strengthened and the disintegrated time was prolonged gradual y. When the ratio of demineralized bone matrix/hyaluronic acid was 15/1, demineralized bone matrix/hyaluronic acid was molded easily, and the disintegrated time was 8 hours in 7 ℃ PBS. The results of cytotoxicity showed that cellproliferation rates were 93.72%, 101.65%and 97.68%at days 1, 4 and 7 respectively, while the cytotoxicity was in grade 0 or 1. These findings indicate that demineralized bone matrix/hyaluronic acid putty at a compound ratio of 15:1 can be molded easily and difficult to disintegrate, with low cytotoxicity.

OBJECTIVETo analyze risk factors, clinical features, outcomes and antibiotic resistance of Escherichia coli(E.coli) causing bloodstream infections in children.

METHODAll inpatients with E. coli positive blood culture in Beijing Children's Hospital from January 2012 to May 2014 were enrolled; 112 cases were included, 66 cases (58.9%) were male, and 46 cases(41.1%) were female. Age range was 2 days to 16 years. Among them, 43 cases (38.4%) were neonates, 19 cases (17.0%) aged from 1 month to 1 year, 14 cases (12.5%) were 1-3 years old, and 36 cases (32.1%) were over three years old. We analyzed the divisions to which the patients were admitted, source of infection, underlying diseases, clinical characteristics, antibiotic resistance, and treatment outcomes, etc.

RESULTForty-six cases (41.1%) were treated in division of hematology, 42 (37.5%) in neonatology, 9 (8.0%) in internal medicine, 8 (7.1%) in surgery, and 7 (6.3%) in pediatric intensive care unit. Sixty-five cases(58.0%) had underlying diseases. Fever was the most frequently presented symptom, as it was seen in 91 cases (81.3%); 52 cases(46.4%) had respiratory symptoms. Among these, 43 cases had pneumonia, 3 cases had respiratory failure, 3 cases were diagnosed as upper respiratory tract infection, 2 had pulmonary hemorrhage and 1 case had bronchitis. Twenty-six cases (23.2%)were diagnosed as severe sepsis and purulent meningitis separately, 14 cases(12.5%) had urinary tract infection. There were 73 (65.2%) strains inducing extended spectrum β-lactamases (ESBLs), of which 6 (8.2%) and 10 (13.7%) strains were resistant to amikacin and carbapenems respectively. Resistance rate against other antimicrobial agents varied from 64.6% to 100%.

OUTCOMES92 (82.1%) cases were cured or had improvement while 20 patients (17.9%) died or could not be cured at the end of treatment. Positive ESBLs (χ(2) = 6.609, P = 0.010), being complicated with severe sepsis (χ(2) = 40.253, P = 0.000) and requiring mechanical ventilation (χ(2) = 34.441, P = 0.000) indicate poor prognosis.

CONCLUSIONPatients with underlying diseases and newborns are susceptible to E. coli bloodstream infection. ESBLs infection, severe sepsis and mechanical ventilation indicate poor prognosis in E. coli blood stream infection. Clinicians may use carbapenems as empirical treatment for ESBLs infection. There may be carbapenem-resistant enterobacteriaceae strains infection if patients receiving treatment with carbapenems have no response.

Adolescent ; Bacteremia ; Carbapenems ; therapeutic use ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Escherichia coli ; drug effects ; Escherichia coli Infections ; blood ; drug therapy ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Meningitis, Bacterial ; Microbial Sensitivity Tests ; Pneumonia ; Urinary Tract Infections ; beta-Lactamases

Objective To explore the clinical effect of Tongluo-Ningxue decoction for hematuria from IgA nephropathy. Methods A total of 60 patients with IgA nephropathy in the hospital between July 2015 and August 2016 were randomly divided into two groups, with 30 cases in each group. The control group was treated with western medicine (Benazepril Hydrochloride plus dipyridamole), whereas the observation group took Tongluo-Ningxue decoction for treatment. The clinical efficacy, clinical symptom score, biochemical index and humoral immune index in the two groups were compared. Results The effective rate was 86.7％ (26/30) in the observation group, and 63.3％ (19/30) in the control group, with the difference between the two groups being statistically significant (χ2=4.356, P=0.037). After treatment, the scores of fatigue and weakness (0.7 ± 0.1 vs. 1.4 ± 0.2, t=18.954), soreness and weakness of waist and knees (0.7 ± 0.1 vs. 1.4 ± 0.2, t=21.071), swollen sore throat (0.6 ± 0.1 vs. 1.4 ± 0.2, t=20.723), and hot flashes and night-time sweating (0.6 ± 01 vs. 1.5 ± 0.2, t=20.732) in the observation group were significantly lower than those in the control group (P<0.05). After treatment, the 24h UP (0.53 ± 0.12 g vs. 0.74 ± 0.15 g, t=5.988), RBC (16.84 ± 5.38 vs. 21.42 ± 9.73, t=2.256), SCr (56.34 ± 7.21 μmol/L vs. 61.27 ± 8.45 μmol/L, t=2.431), and IgA (2.35 ± 0.43 g/L vs. 2.62 ± 0.45 g/L, t=2.376) in the observation group were significantly lower than those in the control group (P<0.05). Conclusions The Tongluo-Ningxue decoction can improve the biochemical indicators, enhance immunity, improve renalfunction and relieve clinical symptoms, and therefore, it is worthwhile for the clinical application.

OBJECTIVETo summarize the clinical features, imaging characteristics, diagnosis and treatment of a case with central nervous system infection caused by Exophiala dermatitidis, as well as to review the related literature.

METHODAssociated literature and clinical data of an 8-year-old boy who was diagnosed as central nervous system infection caused by Exophiala dermatitidis in Beijing Children's Hospital Affiliated to Capital Medical University and hospitalized twice from 2012 to 2014 were analyzed retrospectively.

RESULTThe boy was 8 years old with the chief complaint of dizziness for 2 months, intermittent fever for 1 month accompanied with spasm twice. He was diagnosed as bile ducts space-occupying lesions 2 years ago, when the pathological diagnosis was fungal infection. The boy was treated with irregular anti-fungal therapy. Then the boy developed nervous symptoms, impaired consciousness and abnormal physical activity that developed gradually. After hospitalization the cerebral MRI of the boy showed space-occupying lesions accompanied with edema of surrounding area. Filamentous fungi was found by brain biopsy, which was culture positive for Exophiala dermatitidis. After diagnosis the boy was treated with amphotericin B (AMB), voriconazole and 5-Fu, as well as symptomatic treatment. The state of the boy was improved gradually. Two months later, the boy could communicate with others normally and move personally. The lesions and edema seen on the MRI was decreased moderately. Accordingly, the boy was treated with oral voriconazole maintenance treatment for about 1 year and 4 months after discharge. During this period, the state of him was stable without symptoms. The lesions shown by MRI did not disappear but decreased on regular examination. However, recently the disease of the boy progressed again, with dizziness, neck pain, headache and progressive nervous symptoms (intermittent spasm, inability to cough, and impaired consciousness). The boy died at last, even with the active treatment at the second hospitalization. Exophiala dermatitidis was culture-positive again in his CSF, and was confirmed by PCR successfully.

CONCLUSIONThe central nervous system infection caused by Exophiala dermatitidis is rare. Clinical features of this disease were similar to those of other fungal CNS infection, cerebral MRI of which could show the similar lumpy lesions. Diagnosis of the disease should be based on pathology and culture.

Amphotericin B ; administration & dosage ; Antifungal Agents ; administration & dosage ; Brain ; diagnostic imaging ; microbiology ; pathology ; Central Nervous System Infections ; diagnosis ; drug therapy ; microbiology ; Cerebrospinal Fluid ; microbiology ; Child ; Drug Therapy, Combination ; Exophiala ; isolation & purification ; Fatal Outcome ; Fluorouracil ; administration & dosage ; Humans ; Magnetic Resonance Imaging ; Male ; Mycoses ; diagnosis ; drug therapy ; microbiology ; Radiography ; Voriconazole ; administration & dosage

Vitiligo， a skin pigmentation disorder caused by the loss of melanocytes in the basal layer of the skin， is manifested as creamy white or ivory white pigmented islands on the head， face， hair， areola， genitals， mucous membranes and traumatic areas with distinct borders， seriously affecting the patient’s social， physical， and mental health. The disease has attracted wide attention in the medical circle as a difficult aesthetic dermatosis with an increasing prevalence year by year. There are still blind spots in the hypotheses that autoimmunity， melanocyte autophagy， oxidative stress， autocytotoxicity， neurohumors， and genetic and environmental factors are associated with the pathogenesis of this disease. The commonly used Western medical therapies， including glucocorticoids， small-molecule antagonists， calcium-regulated neurophosphatase inhibitors， biologics， vitamin D derivatives， phototherapy， and surgery are flawed with side effects and prone to recurrence. Traditional Chinese Medicine （TCM） can treat vitiligo via a wide range of pathways and targets， with definite effects and low adverse reactions. Studies have demonstrated that TCM can promote autophagy of melanocytes and protect them from oxidative stress. However， there are few systematic summaries of the signaling pathways in the TCM treatment of vitiligo. Therefore， this paper introduces the main signaling pathways involved in the TCM treatment of vitiligo by reviewing the relevant articles published at home and abroad in recent years. Specifically， the signaling pathways include the molecular hydrogen-activated nuclear factor erythroid-related factor 2 （Nrf2）， tyrosine kinase receptor （c-Kit）， nuclear transcription factor-κB （NF-κB）， Janus tyrosine protein kinase （JAK）， mitogen-activated protein kinase （MAPK）， phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）， and adenosine monophosphate-activated protein kinase （AMPK） signaling pathways.

Enterotoxigenic Escherichia coli（ETEC）infection can induce watery diarrhea，leading to dehydration，electrolyte disturbance，and even death in severe cases. Recombinant B subunit/inactivated whole-cell cholera（rBS/WC）vaccine is effective in preventing ETEC infectious diarrhea. On the basis of the latest evidence on etiology and epidemiology of ETEC，as well as the effectiveness，safety，and health economics of rBS/WC vaccine，National Clinical Research Center for Child Health（The Children’s Hospital，Zhejiang University School of Medicine）and Zhejiang Provincial Center for Disease Control and Prevention invited experts to develop expert consensus on rBS/WC vaccine in prevention of ETEC infectious diarrhea. It aims to provide the clinicians and vaccination professionals with guidelines on using rBS/WC vaccine to reduce the incidence of ETEC infectious diarrhea.