The effect of phenol concentration on the structure and function of microbial communities,which were cultured in different conditions using coking wastewater biofilm as seeding,was investigated by Biolog and denaturing gradient gel electrophoresis(DGGE)methods.The less number of bands of cultivated sam-ples on the denaturing gradient gel electrophoresis fingerprint of 16S rRNA gene indicated reduction of di-versity after enrichment and cultivation.Some bands on the DGGE gel were significantly influenced by the phenol concentration in medium.The results of Biolog showed that the original biofilm sample had the highest substrate utility capacity as measured by average well color development(AWCD).But low concen-tration of phenol enriched sample S7 showed more diverse activity on the utility of Carboxylic acids.The principal component analysis(PCA)of Biolog data revealed that the metabolic patterns were similar when using the same phenol concentration,although the sample S7 much less similar to other cultivated samples.These results suggested that the enrichment and cultivation with phenol supplemented decreased the diver-sity and also changed the metabolic function of the microbial community.Lower phenol concentration in-creased the microbial community metabolic activity.The phenol degrading capacity of isolates from each samples indicated that the enrichment and cultivation condition had changed the type and property of cul-truable bacteria.Based on these results,we concluded that the different microorganisms will be isolated un-der different cultivation condition.

Aorta, Thoracic ; pathology ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; diagnosis ; mortality ; Humans ; Infant ; Infant, Newborn ; Male ; Vascular Malformations ; diagnosis ; mortality

It was expected that recombinant Aspergillus niger glucose oxidase could be expressed in Trichoderma reesei with stable activity. T. reesei CBHI promoter--CBHI ss. gene--A. niger glucose oxidase gene--T. reesei CBHI terminator--A. nidulans gpd promoter--E. coli Hygromycin B phosphotransferase gene--A. nidulans trpC terminator--pUC19 (pCBHGOD) vector was constructed in E. coli DH5alpha by PCR application and gene cloning methods. T. reesei QM9414 protoplast was transformed by T. reesei CBHI promoter-CBHI ss. Gene--A. niger glucose oxidase gene--T. reesei CBHI terminator-A. nidulans gpd promoter--E. coli Hygromycin B phosphotransferase gene--A. nidulans trpC terminator linear DNA fragment (CBHGOD fragment) that was made by digestion of pCBHGOD with Kpn I. T. reesei mutant clone with homologous recombinant A. niger glucose oxidase gene was selected by PCR method. Recombinant glucose oxidase was produced by mutant T. reesei strain under induction of wheat straw for 5 days. Recombinant glucose oxidase molecular mass was showed the same as native A. niger glucose oxidase standard from Sigma company by Western blot analysis. Recombinant glucose oxidase activity was 25u/mL in medium. The yield was 0.5 g/L in comparison with Sigma company glucose oxidase standard. There was no recombinant GOD degradation during Trichoderma reesei cultivation that was showed in Western blot analysis. Trichoderma reesei has capability to be a new recombinant host for Aspergillus niger GOD production.
Aspergillus niger ; enzymology ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Glucose Oxidase ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Trichoderma ; genetics ; metabolism

OBJECTIVETo explore the influence of exogenous putrescine and cadaverine on pro-inflammatory factors in the peripheral blood of rabbits.

METHODSForty ordinary adult New Zealand rabbits were divided into saline, necrotic tissue homogenate (NTH), putrescine, and cadaverine groups according to the random number table, with 10 rabbits in each group. Saline, NTH, 10 g/L putrescine, and 10 g/L cadaverine were respectively peritoneally injected into rabbits of corresponding group in the amount of 1 mL/kg. The blood sample in the volume of 2 mL was collected from the central artery of rabbit ears before injection and at 2, 6, 12, 24, 30, 36, 48, 60 hours post injection (PIH). Contents of TNF-α, IL-1, and IL-6 in the serum were determined with enzyme-linked immunosorbent assay. Data were processed with repeated measurement data analysis of variance and Spearman correlation analysis, and cubic model curve was applied in curve fitting for the contents of inflammatory factors.

RESULTS(1) The serum contents of TNF-α, IL-1, and IL-6 were increased in NTH, putrescine, and cadaverine groups in different degrees at most post injection time points. There was no significant change in the concentrations of the three pro-inflammatory factors in saline group, and they were significantly lower than those of the other three groups at most post injection time points (with F values from 3.49 to 13.58, P values all below 0.05). The serum contents of TNF-α, IL-1, and IL-6 in putrescine group began to increase at PIH 2, 6, and 6, which was similar to the trend of NTH group, but the changes were delayed compared with those of cadaverine group(all at PIH 2). The peak values of TNF-α, IL-1, and IL-6 in putrescine group were respectively (339 ± 36), (518 ± 44), and (265.9 ± 33.5) pg/mL, which were significantly lower than those of cadaverine group [ (476 ± 86), (539 ± 22), and (309.4 ± 27.1) pg/mL], with F values respectively 5.11, 1.90, and 5.56, P values all below 0.05. (2) The period of time in which contents of TNF-α, IL-1, and IL-6 began to increase (PIH 3-4) and the peaking time of the three pro-inflammatory cytokines (PIH 18-30) in putrescine group appeared later than those of cadaverine group (PIH 2 and 12-30). The duration of peaking time of the three pro-inflammatory cytokines in putrescine group was shorter than that of cadaverine group (PIH 18-30 vs. PIH 12-30). The increasing period and the duration of peaking time of TNF-α, IL-1, and IL-6 in putrescine group were close to those of NTH group (PIH 3-5 and 18-30). The correlation coefficient test analysis showed that the trends of changes in contents of three pro-inflammatory cytokines in putrescine group were significantly correlated with those of NTH group (r(TNF-α) = 0.933, P < 0.01; r(IL-1) = 0.967, P < 0.01; r(IL-6) = 0.950, P < 0.01). The obvious correlation between cadaverine group and NTH group was only found in the contents of IL-1 and IL-6 (r(IL-1) = 0.913, P < 0.01; r(IL-6) = 0.883, P < 0.05).

CONCLUSIONSBoth exogenous putrescine and cadaverine can cause inflammatory reaction in rabbits. The trend of the inflammatory reaction induced by putrescine was similar with that by NTH, suggesting that putrescine may play a leading role in the inflammatory reaction induced by necrotic tissue decomposition.

Animals ; Cadaverine ; adverse effects ; Inflammation ; blood ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Necrosis ; blood ; Putrescine ; adverse effects ; Rabbits ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo study the clinical and epidemiologic characteristics and the serum types of enterovirus of hand, foot and mouth disease (HFMD) in children.

METHODSThe RT-nPCR method was established with universal primers within 5' untranslated region of enterovirus and VP1 region of Coxsackievirus A16 (CAV16) and enterovirus 71 (EV 71). Enteroviruses were detected with RT-nPCR in 237 children with HFMD. Clinical and epidemiologic characteristics and serum types of enterovirus of the patients with HFMD were studied.

RESULTSThe patients'age ranged from 7 months to 11 years (mean 4.2 +/- 0.5 years). The majority (94.5%) were less than 6 years old. HFMD was mostly seen in spring and winter (67.9%). Oral mucosal pox or ulcer as well as hand and foot rashes were observed in all 237 patients. Fever occurred in 141 patients (59.5%). Of the 237 patients, 133 (56.1%) were RT-nPCR positive. Of the 133 cases, 38 were positive for EV71, 64 were positive for CAV16, and 31 were negative for both EV71 and CAV16. The patients infected by different types of enteroviruses had similar clinical characteristics. Gene colon and sequence analysis for 12 strains of enteroviruses PCR positive products presented as EV71 (n=5), CAV16 (n=5), ECHO13 (n=1), and CAV5 (n=1).

CONCLUSIONSHFMD tends to occur in younger children less than 6 years old. The majority are affected in spring and winter. EV71 and CAV16 are common pathogens of HFMD. There is no relationship between clinical characteristics and serum types of enteroviruses in HFMD patients.

Child ; Child, Preschool ; Enterovirus ; classification ; Female ; Hand, Foot and Mouth Disease ; virology ; Humans ; Infant ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; Seasons ; Serotyping

Purpose: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen of coronavirus disease 2019. Diagnostic methods based on the clustered regularly interspaced short palindromic repeats (CRISPR) have been developed to detect SARSCoV-2 rapidly. Therefore, a systematic review and meta-analysis were performed to assess the diagnostic accuracy of CRISPR for detecting SARS-CoV-2 infection. Materials and Methods: Studies published before August 2021 were retrieved from four databases, using the keywords “SARS-CoV-2” and “CRISPR.” Data were collected from these publications, and the sensitivity, specificity, negative likelihood ratio (NLR), positive likelihood ratio (PLR), and diagnostic odds ratio (DOR) were calculated. The summary receiver operating characteristic curve was plotted for analysis with MetaDiSc 1.4. The Stata 15.0 software was used to draw Deeks’ funnel plots to evaluate publication bias. Results: We performed a pooled analysis of 38 independent studies shown in 30 publications. The reference standard was reverse transcription-quantitative PCR. The results indicated that the sensitivity of CRISPR-based methods for diagnosis was 0.94 (95% CI 0.93–0.95), the specificity was 0.98 (95% CI 0.97–0.99), the PLR was 34.03 (95% CI 20.81–55.66), the NLR was 0.08 (95% CI 0.06– 0.10), and the DOR was 575.74 (95% CI 382.36–866.95). The area under the curve was 0.9894. Conclusion Studies indicate that a diagnostic method based on CRISPR has high sensitivity and specificity. Therefore, this would be a potential diagnostic tool to improve the accuracy of SARS-CoV-2 detection.

OBJECTIVETo analyze the risk factors of hemorrhagic cystitis after allogeneic hematopoietic stem cell transplantation for beta-thalassemia in children.

METHODSThe clinical records of 30 children with beta-thalassemia undergoing allogeneic hematopoietic stem cell transplantation between December, 2008 and November, 2009 were analyzed.

RESULTSHemorrhagic cystitis occurred in 8 of the 33 patients with an incidence of 24.24%, including 1 with grade I, 6 with grade II and 1 with grade III hemorrhagic cystitis. The median time of hemorrhagic cystitis onset was 22.9 days (range 6-35 days) and the median duration was 11.9 days(range 3-27 days). Univariate analysis indicated that the different types of transplantation and acute graft-versus-host disease affect the occurrence of hemorrhagic cystitis. The children with Allo-PBSCT had higher incidence than those receiving Allo-PBSCT+Allo-UBT and Allo-BMT (P<0.05). The children at an age >or=6 years had obviously higher incidence of hemorrhagic cystitis than those at younger ages.

CONCLUSIONAge is the major factor that affects the occurrence of hemorrhagic cystitis in children undergoing allogeneic hematopoietic stem cell transplantation for beta-thalassemia.

Age Factors ; Child ; China ; epidemiology ; Cystitis ; epidemiology ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Incidence ; Risk Factors ; Transplantation, Homologous ; beta-Thalassemia ; therapy

OBJECTIVETo explore the strategy of damage control in clinical treatment of multiple injuries headed by cervical spinal cord injury.

METHODSA retrospective analysis was performed in 32 patients. Cervical fractures associated with tetraplegia occurred in 18 patients, traumatic intervertebral disk hernia associated with tetraplegia in 2 patients, and cervical fractures and dislocation associated with tetraplegia in 12 patients. Seventeen cases were combined with craniocerebral injury, 7 combined with pulmonary contusion, multi-fractures of rib or hemopneumothorax, 2 combined with pelvic fracture and other 8 combined with fracture of limbs. The neural function was assessed by the American Spinal Injury Association (ASIA) scale.

RESULTSThirty-one patients were followed up for an average of 14 months. Of them, 10 got complete recovery, 13 obtained improvement of more than one ASIA grade, 8 did not improve, and 1 died.

CONCLUSIONSFor the emergency treatment of multiple injuries headed by cervical spinal cord injury, the damage control strategy is the principle to follow. The final operations are preferably performed within 5 to 10 days after injury so as to raise the successful rate of remedy.

Adolescent ; Adult ; Cervical Vertebrae ; injuries ; Emergency Treatment ; Female ; Humans ; Male ; Middle Aged ; Multiple Trauma ; etiology ; surgery ; Quadriplegia ; Retrospective Studies ; Spinal Cord Injuries ; surgery

OBJECTIVEEndomyocardial biopsies from 42 (35 males and 7 females, aged 43.3 years) heart transplant recipients due to end-stage heart failure between June 2004 and January 2006 in our institute were obtained for pathological studies.

METHODSSixteen patients underwent 1 endomyocardial biopsy (right ventricular septum) between 13 days to 5 months, 13 patients underwent second biopsy between 1.5 to 8 months and 10 patients underwent third biopsy between 3 to 8.5 months post transplantation. Specimen were stained by hematoxylin-eosin (HE) and Phosphotungstic Acid Hematoxylin (PTAH) and observed under light microscope and cardiac allograft rejection were evaluated according to the Revision of the 1990 working formulation for the standardization of nomenclature in the diagnosis of heart rejection in 2004.

RESULTSThe rejection grades were as follows: Grade 0 R in 31 biopsies; Grade 1 R (mild rejection 1990 grade 1A, 1B and 2.) in 30 biopsies; Grade 2 R (moderate rejection, 1990 grade 3A) in 3 biopsies; Grade 1 R cellular rejection companies with humoral rejection in 1 biopsy. Cellular rejection with Quilty effect was found in 2 biopsies. Ischemic myocardial injury presented in 4 biopsies. Quilty effect was observed in 1 biopsy. Cytomegalovirus or toxoplasmic myocarditis was not observed.

CONCLUSIONSEndomyocardial biopsy (EMB) is a valuable diagnostic procedure for rejection surveillance in heart allograft recipients. The observed low rejection incidence and mild rejection from specimens of our heart recipients were comparable to the results of developed countries.

Adolescent ; Adult ; Biopsy ; Endocardium ; pathology ; Female ; Heart Failure ; pathology ; surgery ; Heart Transplantation ; Humans ; Male ; Middle Aged ; Myocardium ; pathology ; Postoperative Period

Objective:To investigate the clinical significance of bone metabolism indexes and human leukocyte antigen B27 (HLA-B27)in ankylosing spondylitis(AS)and analyze their diagnostic value. Methods:The subjects were 94 cases of AS patients,239 cases with other diseases and 80 healthy controls, and the results were retrospectively surveyed. Results:In the AS group, the concentration of the β-collagen specific sequence (β-CTX ) was higher ( P<0. 05 ) while the concentrations of osteocalcin ( OC ) , 25-hydroxyvitamin D[25-(OH)D] and parathyroid hormone(PTH)were lower than those in the control group. In the AS group,there was a positive correlation between the concentration of C-reactive protein(CRP)and β-CTX(P<0. 01),and the concentration of CRP was negatively correlated with the concentration of 25-(OH)D(P<0. 05). However,the concentrations of other bone metabolic indexes had no correlation with the concentration of CRP(P<0. 05). The positive rate of HLA-B27 in the AS group was significantly higher than that in other groups(P<0. 05),and HLA-B27 had highly sensitive and specific in the diagnosis of AS. In the AS group,the sensitivity of the concentration of 25-( OH) D was higher than β-CTX,while its specificity was lower than β-CTX. Conclusion:Bone metabolic indexes have great value in early screening and clinical diagnosis of AS,especially β-CTX and 25-( OH) D were more obvious.