Objective To explore the change and value of interleukin (IL)-2,IL-6 and tumor necrosis factor (TNF)-α in the serum samples of patients with brucellosis.Methods The levels of serum IL-2,IL-6 and TNF-α were measured using the enzyme linked immunosorbent assay (ELISA) method in 155 patients with brucellosis in different clinical stages (including 69 cases in acute,34 cases in subacute and 52 cases in chronic periods) and 50 healthy controls.IL-2 was used to represent the helper T cell (Th)-type 1 cytokines,IL-6 represent Th2 type cytokines,and the Th1/Th2 ratio in different clinical stages was compared.Results The expression level of serum IL-2 in patients with brucellosis was different (ng/L:acute:10.15 ± 2.01;subacute:9.53 ± 1.68;chronic:6.76± 1.31;control:47.25 ± 5.68),the differences were statistically significant (F =74.921,P ＜ 0.05).The expression levels of serum IL-2 in acute,subacute and chronic periods of brucellosis patients were significantly lower than that of healthy controls (all P ＜ 0.05).The expression levels of serum IL-2 in acute and subacute periods of brucellosis patients were higher than that of chronic period patients (all P ＜ 0.05).The expression of IL-6 and TNF-α in serum of patients in different clinical stages were different (ng/L:acute:615.22± 341.07,802.55 ± 479.53;subacute:478.45 ± 105.33,680.21 ± 366.95;chronic:306.37 ± 96.12,455.36 ± 176.27;control:121.45 ± 30.16,87.51 ± 24.03),the differences were statistically significant (F =57.692,63.210,all P ＜ 0.05).The levels of serum IL-6 and TNF-o were significantly higher in patients with brucellosis in acute,subacute and chronic clinical stages than that of healthy controls (all P ＜ 0.05).Serum IL-6 and TNF-α levels in acute period of brucellosis patients were higher than those of subacute and chronic periods patients (all P ＜ 0.05);the levels of serum IL-6 and TNF-α in subacute period of brucellosis patients were also significantly higher than that of chronic period patients (all P ＜ 0.05).The Th1/Th2 ratio was different (acute:0.02 ± 0.00;subacute:0.02 ± 0.00;chronic:0.23 ± 0.02;control:0.41 ± 0.06) in clinical patients with brucellosis,the differences were statistically significant (F =17.843,P ＜ 0.05),the value of Th1/Th2 in acute and subacute period patients were lower than that of chronic period (all P ＜ 0.05).Conclusion IL-2,IL-6 and TNF-α may play an important role in the pathogenesis of brucellosis,and their dynamic changes can reflect the progression and outcome of brucellosis.

Objective To discuss the clinical outcomes of old fracture-dislocation of proximal interphalangeal joint treated with a self-made distraction external fixator which consists of 3 pins and rubbers.Methods From March 2005 to August 2011,8 cases of old fracture-dislocation of proximal interphalangeal joint treated with a pins-rubber distraction external fixator were retrospectively reviewed.There were 6 males and 2 females,aged from 17 to 58 years,averaged 23.2 years.The middle finger was involved in 5 cases and ring finger was involved in 3 cases.The right hand was affected in six patients,and the left hand was affectd in two patients.The mechanism of injury was sports injury in 6 cases,traffic injuries in 1,and sprain in 1.All of the cases were closed injury.The mean time from injury to surgery was 58 d (range,45-90 d).According to Schenck classification of the proximal interphalangeal joint fracture-dislocation,there were 4 cases of type Ⅰ,3 of type Ⅱ,I of type Ⅲ.Based on the Michigan Hand Outcomes Questionnaire (MHQ) for subjective hand function score,the average preoperative score was 56; the range of motion of proximal interphalangeal joint was 0°.Grip strength was 60％-80％ of the contralateral hand.Results The follow-up lasted 6-51 months,with a mean period of 20.5 months.Solid bone healing was eventually achieved in all patients.The average healing period was 3 months (range,2-5 months).The mean normalized MHQ score was 88 (range,84-92).Grip strength was 85％-100％ of the contralateral hand.Extension of proximal interphalangeal joint was -10°-0° (average,-5°).Flexion angle was 75°-100° (average,89.4°).Postoperative complications included slight joint space narrowing (2 cases),joint incongruity (＜ 1 mm) (2 cases) and fusiform arthrocele (2 cases).No infection,pin loosening,and pin breakage was encounted.Conclusion A pins-rubber distraction external fixator is a minimally invasive,reliable and effective treatment for old fracture-dislocation of proximal interphalangeal joint.

Objective An experiment study on the femoral head plastic operation to prevent and treat aseptic necrosis of the femoral head. Methods 60 Chinese white rabbits were equally random divided into 4 groups. The four groups except A group were made a globose defect and filled with 95% alcohol tampon for 30 minutes. The B group was natural repair group. The C group was filled with bone cement after being made defect. The D group was filled with bone cement added barium sulfate agent after being made defect. After 12 weeks, all rabbits in four groups were killed. The specimens were random divided into two parts. The articular cartilage was observed and measured immediately. The other specimen was determined with histological examination and extreme anti-pressure rigidity test . Result The femoral heads of bone cement group and the femoral heads of contrast agent bone cement group kept their outline all the time through the articular cartilage observation. After 12 weeks the cartilage thickness of natural repair group (511.74 ± 69.00) was thinner than the other three groups [ control group ( A group ) (511.74 ± 69 00 );Bone cement group ( C group) (468. 36 ± 82. 99 ); Bone cement group contrast agent ( D group ) ( 515.61 ±64. 65 ) ], and it had significant difference ( P < 0. 05, P < 0. 01 ). The rigidity of natural repair group [ (676. 57 ±50. 92) N/mm] had significant difference with that of the other three groups[ A group( 884. 66±52. 29)N/mm;C group(1178.40 ± 170.45) N/mm;D group(928.60 ± 104.42) N/mm] ( P <0.05, P<0. 01 ). Conclusions Femoral head plastic operation was simple and it could release pain. And femoral head outline could be kept integrity. It would be a new operation which was used to treat young patients in Ficat Ⅱ～Ⅲ period to postpone or substitute total hip replacement.

Sanqi is a popular traditional Chinese medicine and commonly used for promoting blood circulation and removing blood stasis. Notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and Rd are the major active constituents of Sanqi. The purpose of the study was to investigate the pharmacokinetic behavior of the five active constituents from total saponin from Sanqi when it was used in the blood stasis animals or in combination with Gegen. The concentrations of the five active constituents in rat plasma were determined by an ultra-HPLC-ESI-MS/MS method. The main pharmacokinetic parameters were calculated and statistically analyzed using the unpaired student's t-test. It was found that the pharmacokinetic parameters of notoginsenoside R1, ginsenoside Rg1 and Rb1 represented a statistically significant difference (Po0.05) between the normal rats and the blood stasis rats after administration of total saponin from Sanqi (TSFS). And there were statistically significant differences (Po0.05) in the pharmacokinetic parameters of all the five constituents between administration of TSFS alone and combined with total flavonoid from Gegen (TFFG) in blood stasis rats. It suggested that the pharmacokinetic behavior of the active constituents from TSFS could be changed when it was used in blood stasis animals or in combination with TFFG.

Objective This study was designed to compare the mRNA expression of Toll-like receptors 2 (TLR2) and Toll-like receptors 4 (TLR4) in monocytes between brucellosis cases and healthy controls,to explore the role and value of TLR2 and TLR4 in the pathogenesis of brucellosis.Methods From March to June in 2015,a total of 155 brucellosis cases at different clinical stages were diagnosed in Ulanqab Center for Endemic Disease Prevention and Control and 50 healthy controls were selected as research subjects.Subjective clinical symptoms of all patients were observed.Serological detection of all cases was done using serum tube agglutination test (SAT).The mRNA expression of TLR2 and TLR4 in peripheral blood cell was detected by real-time PCR technique.Results Fever,hyperhidrosis,weak,bone ache and hepatosplenomegaly were found in patients of acute (69 people),subacute (34 people) and chronic (52 people) periods.The incidences of fever and hyperhidrosis in acute and subacute periods were higher than that in chronic period,and the differences were statistically significant (x2 =58.427,26.190,all P ＜ 0.01).The incidence of bone ache in chronic period was higher than that in acute period,and the difference was statistically significant (x2 =9.264,P ＜ 0.01).In the agglutination titer of 1:800 and higher,the positive rate of chronic period was lower than those in acute and subacute periods,and the differences were statistically significant (x2 =14.302,8.682,all P ＜ 0.01).The mRNA expression levels of TLR2 and TLR4 were different among different clinical periods of brucellosis patients,and the differences were statistically significant (F =17.502,24.931,all P ＜ 0.01).The expression levels of TLR2 mRNA (8.67 ± 2.39) and TLR4 mRNA (12.38 ± 3.87) in acute period of brucellosis patients were higher than those of subacute period (5.21 ± 1.76,7.62 ± 2.21),chronic period (1.25 ± 0.47,1.72 ± 0.55) and healthy control (1.17 ± 0.23,1.43 ± 0.62),and the differences were statistically significant (all P ＜ 0.01).The expression levels of TLR2 and TLR4 mRNA in subacute period of brucellosis patients were higher than those of chronic period and healthy controls,and the differences were statistically significant (all P ＜ 0.01).The expression levels of TLR2 and TLR4 mRNA were not significantly different (all P ＞ 0.05) between chronic period and healthy controls.Conclusion TLR2 and TLR4 may be the specific recognizing receptors participated in the immune response in brucellosis and associated with the occurrence and development of brucellosis.

Endotoxin removal is essential for the safety of biological products. To remove endotoxin efficiently, we used polymyxin B (PMB) affinity adsorbent to remove endotoxin from protein solutions by static adsorption. We studied the effects of spacer length and ligand density of the affinity adsorbent, pH, salt type and concentration, protein type and concentration, endotoxin concentration, and additive on endotoxin removal and protein recovery. Endotoxin content and protein concentration were determined by test and Lowry assay respectively. The results showed that PMB affinity adsorbent had high capacity, high adsorption speed, high removal efficiency and good reusability. In addition, ligand density, pH, salt concentration and the isoelectric point and hydrophobicity of protein all had remarkable influence on the endotoxin removal. Under the optimal conditions, the recoveries of hemoglobin, human serum albumin and lysozyme were 87.2%, 73.4% and 97.3%, respectively, and the corresponding endotoxin removal rates 99.8%, 97.9% and 99.7%, respectively. This study illustrated the effects of solution conditions on the efficiency of endotoxin removal and protein recovery, and would provide useful reference for the efficient removal of endotoxin from biological products.
Adsorption ; Chromatography, Affinity ; methods ; Drug Contamination ; prevention & control ; Endotoxins ; isolation & purification ; Hemoglobins ; isolation & purification ; Polymyxin B ; chemistry ; Proteins ; isolation & purification ; Pyrogens ; isolation & purification ; Serum Albumin ; isolation & purification ; Solutions

Escherichia coli strain DC1515, deficient in glucose phosphotransferase (ptsG), lactate dehydrogenase (ldhA) and pyruvate:formate lyase (pflA), is a promising candidate for the fermentative production of succinate. To further improve the succinate producing capability of DC1515, we constructed plasmid pTrchisA-pyc with heterogenous pyruvate carboxylase (pyc) from Bacillus subtilis 168 under the Trc promoter and introduced it into DC1515. We used lactose as a substitute of IPTG to induce pyc. We optimized the culture conditions such as the lactose addition time, the lactose concentration and the culture temperature after induction for succinate production. We also explored the effect of lactose supplement during the fermentation. The results showed that pyc can be expressed under lactose induction in the fermentative medium with 15 g/L glucose due to the deficient of ptsG in DC1515. Under optimized conditions, the final succinate concentration reached to 15.17 g/L, which was 1.78-fold higher than that of control strain. If complementing lactose twice to the concentration of 1 g/L during the fermentation, the final succinate concentration could further reach to 17.54 g/L. This work might provide valuable information for gene expression in E. coli strains using lactose as inducer for succinate production in a glucose-medium. Due to the reduced cost, E. coli is becoming a more promising strain for succinate production through fermentation.
Bacillus subtilis ; enzymology ; Culture Media ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Lactose ; pharmacology ; Promoter Regions, Genetic ; Pyruvate Carboxylase ; biosynthesis ; genetics ; Succinic Acid ; metabolism

Objective:To investigate the effect of hemoglobin on Shp2 expression in brain tissues and adherens junction in blood brain barrier in rats after cerebral hemorrhage.Methods:Eighty male SD rats were randomly allocated into sham-operated group and groups of cerebral hemorrhage for 6 h, 24 h, 3 d, and 7 d ( n=16). Rat models in the groups of cerebral hemorrhage were established by intracerebral injection of 20 μL of hemoglobin. Six h, 24 h, 3 d, and 7 d after injection, neurological functions were assessed by Longa scale; brain water content and brain organ coefficient in the harvested cerebral tissues were calculated by wet-dry weighting method; Shp2 mRNA expression was analyzed by real-time fluorescence quantification reverse transcription PCR (qRT-PCR); Shp2 positive neurons was detected by immunohistochemistry; the protein expressions of Shp2, α-catenin, β-catenin and vascular endothelial (VE)-cadherin, and phosphorylated (p-) α-catenin, p-β-catenin and p-VE-cadherin were explored by Western blotting. Results:As compared with those in the sham-operated group, Longa scale scores in groups of intracerebral hemorrhage for 6 h, 24 h, 3 d and 7 d were significantly increased ( P<0.05). As compared with the sham-operated group, groups of intracerebral hemorrhage for 24 h, 3 d and 7 d had significantly increased brain water content and brain organ coefficient, statistically reduced Shp2 mRNA expression, statistically samller amount of Shp2 positive neurons, and statistically reduced Shp2 protein expression, and significantly increased protein expressions of p-α-catenin, p-β-catenin and p-VE-cadherin ( P<0.05). Conclusion:Intracerebral injection of hemoglobin can downregulate Shp2 expression and promote phosphorylation of adherens junction proteins, therefore induce the disruption of adherens junction, which might be a critical mechanism of blood-brain barrier disruption and brain edema.

Objective·To compare the clinical features between different subtype bipolar patients with first mania episode, and to contribute to early identification of bipolar disorder. Methods·This study was based on the database named as National Bipolar Mania Pathway Survey (BIPAS). From November 2012 to January 2013, bipolar patients from 26 mental health facilities in China were enrolled in current study. The clinical features were compared between mania patients of different subtypes, including hypomania (groupⅠ), mania without psychotic symptoms (groupⅡ), mania with psychotic symptoms (group Ⅲ) and mixed state (group Ⅳ). Results·There was significant difference in the percentage of clinical symptoms between different subtype bipolar patients with first mania episode, especially the mania and anxiety related symptoms. Group Ⅰ, Ⅲ , Ⅳ were further compared with groupⅡ, which was considered as the typical bipolar disorder. The results showed that the mania related symptoms was significantly higher in group Ⅱ, but anxiety related symptoms was significantly higher in group Ⅰ, Ⅲ, Ⅳ. Moreover, Logistic regression analysis revealed that more eloquent or humor and unusually restless could be in favor of the diagnosis of hypomania; younger and mania or hypomania as first episode might be in favor of the diagnosis of mania with psychotic symptoms; older, national minorities and unusually restless could be in favor of the diagnosis of mixed state. Conclusion·The clinical features between different subtype bipolar patients with first mania episode are various, and analysis of the clinical features can contribute to early identification of bipolar disorder.

Objective To investigate the effect of suction assisted closure (SAC) in treatment of children's leukemia with refractory ulcers.Methods The clinical data of 15 children who had leukemia associated with refractory ulcers from January 2013 to April 2017 in Department of Orthopedics,Wuhan Children's Hospital,Tongji Medical College of Huazhong University of Science & Technology were retrospectively analyzed.There were 8 males and 7 females,among whom 9 cases were acute non-lymphocytic leukemia and 6 cases were acute lymphoblastic leukemia.Patient's ages ranged from 2 to 9 years old with the mean age of 6.2 years.There were 5 cases of refractory ulcers located in the medial side of thigh and 4 cases in the perineum and around anus,3 cases in the ankle,2 cases in the crus and 1 case in the lateral side of thigh.The refractory ulceration area varied from 2 cm × 3 cm-7 cm × 5 cm,and the depth varied from 0.6-1.8 cm.All the 15 cases were cultured with secretions,including 7 cases of Staphylococcus aureus,4 cases of Escherichia coli,2 cases of Klebsiella pneumoniae,1 case of Pseudomonas aeruginosa,and 1 case with no bacteria detected.These children were transferred from Department of Hematology to Department of Orthopedics when there was no improvement in conventional therapy with the average treatment of 26 d (11-36 d).All of these children underwent SAC after thorough debridement.Secondary suturing of ulcers closure was performed or skin transplantation according to the recovery condition of ulcers.Results All the 15 patients of refractory ulcers achieved fresh granulation tissue after SAC treatment.Ten of them who had refractory ulcers could be sutured directly after once SAC treatment,and 5 cases who had refractory ulcers underwent SAC treatment again,in which 3 cases could be sutured directly,and the other 2 cases needed to reduce the wound area and autologous skin graft.All of these patients had refractory ulcers healed after SAC treatment,without any local or systemic complications.Conclusions SAC can bring the deep refractory ulcers into full drainage,protect the refractory ulcers against contamination,stimulate the growth of granulation tissue.SAC is a simple and effective method in treatment of children's leukemia with refractory ulcers.