In order to explore the effect of peptidoglycan hydrolase on the viable cell counts of Bacillus amyloliquefaciens and the yield of alkaline protease, five peptidoglycan hydrolase genes (lytC, lytD, lytE, lytF and lytG) of B. amyloliquefaciens TCCC111018 were knocked out individually. The viable cell counts of the bacteria and their alkaline protease activities before and after gene deletion were determined. The viable cell counts of the knockout mutants BA ΔlytC and BA ΔlytE achieved 1.67×10⁶ CFU/mL and 1.44×10⁶ CFU/mL respectively after cultivation for 60 h, which were 32.5% and 14.3% higher than that of the control strain BA Δupp. Their alkaline protease activities reached 20 264 U/mL and 17 265 U/mL, respectively, which were 43.1% and 27.3% higher than that of the control strain. The results showed that deleting some of the peptidoglycan hydrolase genes effectively maintained the viable cell counts of bacteria and increased the activity of extracellular enzymes, which may provide a new idea for optimization of the microbial host for production of industrial enzymes.
Bacillus amyloliquefaciens/genetics* ; Bacterial Proteins ; Cell Count ; Endopeptidases/genetics* ; N-Acetylmuramoyl-L-alanine Amidase/genetics*

Objective:To analyze the clinical characteristics and efficacy of drug treatment in children with inflammatory bowel disease (IBD) at different ages of onset.Methods:The clinical data of 87 children with IBD admitted to Department of Gastroenterology in Children′s Hospital, Capital Institute of Pediatrics from January 2009 to December 2018 were collected. The patients were divided into four groups according to the age of onset: 0 -<2 years old group (36 cases), 2 -<6 years old group (10 cases), 6 -<10 years old group (12 cases) and 10 -<18 years old group (29 cases). The clinical manifestations, laboratory examination, endoscopic findings, pathologic and genetic changes, and treatment were compared among different age groups with chi-square test or Fisher′s exact text.Results:(1) A total of 87 patients were diagnosed with IBD, including 50 Crohn′s disease (CD) (57%), 25 ulcerative colitis (UC) (29%) and 12 unclassified inflammatory bowel disease (IBD-U) (14%). (2) Patients with fever accounted for 78% (28/36) and 8/10 in the 0 -<2 years old group and 2 -<6 years old group, respectively. Patients with abdominal pain and perianal diseases accounted for 6% (2/36) and 47% (17/36) in the 0 -<2 years old group, and their proportions were significantly different among the four groups (χ 2=8.369, 40.317 and 13.130, all P<0.05). (3) Leukocytosis, thrombocytosis and anemia were more common in the 0-<2 years old group, seen in 72% (26/36), 31% (11/36) and 81% (29/36), respectively. There were significant differences in the changes of complete blood count among the four groups (χ 2=21.919, 8.095 and 11.520, all P<0.05). (4) Colonic involvement accounted for 85% (17/20) in the 0 -<2 years old CD patients. While in the CD patients over 6 years old, 61% (14/23) had inflammation of ileum and colon, with a significant difference compared to that in patients under 6 years old (19% (5/27) , χ 2=9.455, P=0.003). Also, the location of bowel inflammation among the four groups were significantly different (χ 2=21.120, P<0.01). (5) Noncaseating granulomas were found in 15 (30%) CD patients, and crypt abscess was found in 11 (44%) UC patients. (6) Among the 24 patients whose genes were analyzed by high throughput sequencing, 12 had pathogenic single gene mutation. (7) There were 25 patients treated with total enteral nutrition. Among the 25 patients treated with thalidomide, 20 (80%) had clinical remission or partial remission. Among the 19 CD patients treated with infliximab (IFX), 14 had clinical remission at the 6 th week of treatment, and the proportion of remission maintenance at the 30 th week of treatment was 12/14. (8) The rate of clinical remission or partial remission was 64% (23/36) in the 0 -<2 years old group, 8/10 in the 2 -<6 years old group, 11/12 in the 6 -<10 years old group, and 83% (24/29) in the 10 -<18 years old group. Conclusions:The proportion of CD was higher than that of UC in this study. Infant onset inflammatory bowel disease was more likely to present with perianal lesions, and was usually associated with leukocytosis, thrombocytosis and anemia, and has high possibility of single gene mutation. IFX may be effective in treating CD.

Objective: To analyze the clinical and genotypic characteristics of infantile inflammatory bowel disease (IBD). Methods: The age of onset, family history, clinical manifestations, and treatment effect were retrospectively analyzed in 39 infants (male 23 cases, female 16 cases) with IBD who were admitted to the Department of Gastroenterology in Children′s Hospital, Capital Institute of Pediatrics from January 2007 to December 2017. Next generation sequencing (NGS) based on target gene panel was used for gene analysis in 17 patients. Results: The median age of onset was 0.5 (0.5, 1.0) month. The most common clinical symptoms included diarrhea (39, 100%), malnutrition (38, 97%), hematochezia (34, 87%), fever (25, 64%), and perianal diseases (24, 61%). Four children had associated family history. Among the 17 patients whose gene was analyzed, 10 were found to have the pathogenic gene variation, within whom 7 had interleukin-10 receptor α subunit (IL-10RA) mutation, 2 had CYBB heterozygous mutation, 1 had interleukin-10 receptor β subunit (IL-10RB) mutation. The therapeutic medicine included mesalazine, steroids, and thalidomide. Eighteen children (46%) reached clinical remission (10 cases) or partial remission (8 cases). Conclusions The incidence of single gene mutation in infants with IBD is high, with IL-10RA mutation as the most common. Refractory diarrhea and malnutrition may indicate infantile IBD.

Objective To analyze the clinical and genotypic characteristics of infantile inflammatory bowel disease (IBD). Methods The age of onset, family history, clinical manifestations, and treatment effect were retrospectively analyzed in 39 infants (male 23 cases, female 16 cases) with IBD who were admitted to the Department of Gastroenterology in Children′s Hospital, Capital Institute of Pediatrics from January 2007 to December 2017. Next generation sequencing (NGS) based on target gene panel was used for gene analysis in 17 patients. Results The median age of onset was 0.5 (0.5, 1.0) month. The most common clinical symptoms included diarrhea (39, 100%), malnutrition (38, 97%), hematochezia (34, 87%), fever (25, 64%), and perianal diseases (24, 61%). Four children had associated family history. Among the 17 patients whose gene was analyzed, 10 were found to have the pathogenic gene variation, within whom 7 had interleukin‐10 receptor α subunit (IL‐10RA) mutation, 2 had CYBB heterozygous mutation, 1 had interleukin‐10 receptor β subunit (IL‐10RB) mutation. The therapeutic medicine included mesalazine, steroids, and thalidomide. Eighteen children (46%) reached clinical remission (10 cases) or partial remission (8 cases). Conclusions The incidence of single gene mutation in infants with IBD is high, with IL‐10RA mutation as the most common. Refractory diarrhea and malnutrition may indicate infantile IBD.

Hydroxy amino acids, constituents of chiral pharmaceutical intermediates or precursors, have a variety of unique functions in the research fields of biotechnology and molecular biology, i.e. antifungal, antibacterial, antiviral and anticancer properties. Biosynthesis of hydroxy amino acids is preferred because of its high specificity and selectivity. The hydroxylation of hydrophobic amino acids is catalyzed by hydroxylase, which belongs to the mononuclear non-heme Fe(Ⅱ)/α-ketoglutarate-dependent dioxygenases (Fe/αKGDs). Fe/αKGDs utilize an (Fe(Ⅳ)=O) intermediate to activate diverse oxidative transformations with key biological roles in the process of catalytic reaction. Here, we review the physiological properties and synthesis of hydroxy amino acids, especially for the 4-HIL and hydroxyproline. The catalytic mechanism of Fe/αKGDs is elucidated, and the applications of hydroxy amino acids in industrial engineering are also discussed.

Objective: To analyze the clinical features and interleukin-10 receptor gene mutations in six infants with very early onset inflammatory bowel disease (VEO-IBD). Methods: Four girls and two boys with VEO-IBD admitted to Children's Hospital Affiliated to Capital Institute of Pediatrics from June 2016 to September 2017 were reviewed. The clinical data including general condition, clinical symptoms, laboratory tests, and colonoscopy and pathological results were collected and analyzed. Interleukin-10 receptor α subunit (IL-10RA) gene was examined in all patients. Results: Persistent diarrhea and fever were the most common symptoms and were found within 1 month after birth in all 6 patients. Anemia, oral ulcer or perianal lesions and growth retardation were common concomitant symptoms. All patients had colonoscopy examination and the results showed multiple ulcers affecting the colon with biopsies revealing acute and chronic inflammation. Three patients were found to have cryptitis and crypt abscesses. Gene sequencing revealed IL-10RA gene mutations in all six patients, including 3 cases with homozygous mutations (one with c.537G>A and two with c.301C>T) and 3 heterozygous mutations (paternal c.301C>T in all cases; maternal c.299T>G, c.350G>A and c.537G>A, respectively) . After conventional treatment, one got clinical and pathological improvement according to colonoscopy, three improved clinically, one worsened and died, and one died of septic shock secondary to intestinal perforation. Conclusions VEO-IBD is associated with IL-10RA mutation, usually with severe intestinal symptoms and significant extra-intestinal symptoms, as well as varied responses to conventional treatment. In our study, c.301C>T and c.537G>A are the most common mutations.

Objective To study the changes of activities of phase Ⅰ and Ⅱ drug-metabolic enzymes in the spleen of rats with spleen-kidney yang deficiency syndrome.Methods The rat model of spleen-kidney yang deficiency syndrome was established by gastric gavage of Radix et Rhizoma Rhei decoction combined with injection of hydrocortisone for 17 continuous days.And then we detected the activities of 6 kinds of phase Ⅰ drug-metabolic enzymes of CYP2C19,CYP2D6,CYP2C9,CYP1A2,CYP2C8,CYP3A4,and 4 kinds of phase Ⅱ drugmetabolic enzymes of phenol sulfotransferase (PST),uridine diphosphate glucuronosyl transferase 1 (UGT1),glutathione transferase (GST),estrogen sulfotransferase (SULT1E1) in the spleen.Results Compared with the normal control group,the activities of PST,UGT1,GST and SULT1E1 in the model group were significantly decreased (P ＜ 0.05 or P ＜ 0.01),and the activity of CYP1A2 was significantly increased (P ＜ 0.01),while CYP2C19,CYP2D6,CYP2C8,CYP3A4,CYP2C9 enzymes showed no obvious changes(P ＞ 0.05).Conclusion The activities of splenic drug-metabolic enzymes,in particular the phase Ⅱ enzymes,are significantly varied under the state of spleen-kidney yang deficiency.

Objective To investigate the effects of hemin on the quantity and apoptosis of human umbilical cord blood-derived late endothelial progenitor cells (EPCs) in vitro.Methods Mononuclear cells were isolated from human cord blood by density gradient centrifugation and were induced to differentiate to late EPCs in vitro.The second to third generation of attached late EPCs in good state were randomly plated for 24 h under different concentrations(0, 5, 10, 15 and 20 μmol/L)of hemin.Cell viability and proliferation were measured with typan blue staining and cell counting kit-8, respectively.Cell adhesion was analyzed by adhesive assay, and cell apoptosis was detected by flow cytometry.Results Compared to control group, hemin promoted viability of late EPCsat lower concentrations(5and 10μmol/L).Meanwhile, proliferation and adhesion were also improved and apoptosis was inhibited when the concentrations of hemin were 5,10, or 15 μ mol/L.All these effects were most prominent when hemin concentration was 10 μmoL/L, while the effects above were reversed when hemin concentration was moderated to 20 μmol/L.In addition, hemin showed a time-dependent manner in promoting cell proliferation and adhesion, and inhibiting apoptosis.That effects were most obvious at 24 h.Conclusions Lower concentration of herin augments the quantity and adhesion of late EPCs, inhibits cell apoptosis, while higher concentration present the reversed effects.

OBJECTIVETo explore the effective methods for the treatment of complex tibial plateau fractures.

METHODSFrom May 2008 to April 2011, 28 patients with complex tibial plateau fractures were treated indirect reduction techniques, bilateral locking plate fixation combined with autologous bone grafts. There were 21 males and 7 females, with an average age of 43 years ranging from 21 to 65. There were 11 cases in Schatzker type V, 17 in VI. The effect was evaluated by Rasmussen standard on clinical and radiological.

RESULTSAll patients were followed-up for 7 to 36 months (averaged of 21.5 months). Healing time of fracture was from 3 to 8 months (averaged 5.5 months). The results of Rasmussen scores in clinical was 4.50 +/- 1.32 in pain, 4.32 +/- 1.63 in walking ability, 4.07 +/- 1.34 in knee activity, 4.78 +/- 1.27 in stability of the knee, 4.85 +/- 1.12 in stretch knee; the results in radiation was 5.07 +/- 0.92 in articular surface collapse, 5.00 +/- 0.98 in platform widened, 5.14 +/- 0.85 in knee external varus. The effect result was excellent in 8 cases, good in 15, fair in 3 and poor in 2.

CONCLUSIONThe key for the treatment of complex tibial plateau fractures was to fully assess the damage as much as possible to protect the soft tissue, select the appropriate timing of surgery and surgical incision, application of indirect reduction techniques, limited incision and effective internal fixation to restore joint surface smooth and good limb alignment, early exercise, in order to achieve maximum recovery of joint function.

Adult ; Aged ; Bone Plates ; Bone Transplantation ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Humans ; Male ; Middle Aged ; Recovery of Function ; Retrospective Studies ; Tibial Fractures ; diagnostic imaging ; physiopathology ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

Objective To study the mechanism of oxidative damage in myocardial tissue after limb ischemia reperfusion (IR), and the protective effects of heme oxygenase-1 on myocardial injury in experimental rats. Method The models of bilateral hind limbs ischemia and reperfusion in rats were established by using tourniquets applied to the roots of both hind limbs until palm blanched and pulseless for 4 hours. A total of 56 SD rats were randomly (random number) divided into 7 groups, namely one normal control group ( n = 8) and 6 ischemia-reperfusion groups as per different lengths of reperfusion time, e. g. 2 hrs, 4 hrs, 8 hrs, 16 h rs and 24 hr ( n = 8 each). The experimental rats were sacrificed after different lengths of reperfusion time. Specimens of myocardium and blood were taken for assays of malonaldehyde (MDA), superoxide dismutase (SOD) and myeloperoxidase (MPO), and pathological changes of myocardium were observed, and the expressions of HO-1 mRNA in myocardium were detected. Data were analyzed with ANOVA. Results (1) Compared with the control group, the levels of serum MDA and myocardial MDA of rats were increased in all IR groups and were higher (P < 0.05), and the levels of MDA reached the peak after reperfusion for 4 hours. The levels of serum SOD and myocardial SOD in rats of all IR groups were decreased and lower than those in rats of the control group ( P < 0.05), and the levels of serum SOD dropped away to the lowest point after reperfusion for 4 hours, and the levels of myocardial SOD fell off to the bottom after reperfusion for 8 hours. The levels of serum MPO and myocardial MPO were significantly increased in rats of all IR groups compared with the control group (P < 0.05). The levels of serum MPO reached peak after reperfusion for 4 hours, and the levels of myocardial MPO were increased to the highest spot after reperfusion for 6 hours. (2) The pathological changes in myocardium showed the most severe damage after reperfusionfor 4-6 hours.(3) After reperfusion for 2 hours, there were no significant differences in the expression of HO-1 mRNA between IR groups and control group (P >0.05), and after reperfusion for 4 hours and over, the expressions of HO-1 mRNA were markedly increased in IR groups and reached peak after reperfusion for 16 hours in comparison with the control group (P < 0.05). Conclusions The activation of neutrophils and free radicals may play a primarily adverse role in myocardial injury after limb IR, and the increase in the expression of HO-1 mRNA lessens the harm effects of IR on myocardium.