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 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.

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:To observe the effects of preventative moxibustion on analgesia,substance P(SP),prostaglandin(PG)F2α and PGE2 in rats with dysmenorrhea due to cold-dampness stagnation,and to explore the analgesic mechanism. Methods:Sixty-four female Wistar non-pregnant rats were randomly divided into a blank group,a model group,a Western medicine group,and a preventative moxibustion group,with 16 rats in each group.Eight qualified diestrus rats were selected from each group.Except for the blank group,the other three groups established models of dysmenorrhea due to cold-dampness stagnation using an ice water bath combined with estradiol benzoate and oxytocin.On the 8th day after modeling,the preventative moxibustion group was treated with gentle moxibustion at Shenque(CV8)and Guanyuan(CV4),and the Western medicine group was given ibuprofen solution for 4 consecutive days.On the 11th day,the intervention groups(i.e.the Western medicine group and the preventative moxibustion group)were treated once again after being injected with oxytocin.The writhing score and the pain threshold of rats were determined;the serum levels of brain-derived neurotrophic factor(BDNF),SP,PGF2α,and PGE2 were measured;the mRNA and protein expression levels of BDNF and its receptor tropomyosin receptor kinase B(TrkB)in the spinal dorsal horn and hypothalamus were detected. Results:Compared with the blank group,the writhing score increased(P<0.01),the pain threshold decreased(P<0.01),the serum levels of BDNF,SP,and PGF2α increased(P<0.01),while the PGE2 decreased(P<0.01);the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus increased(P<0.01)in the model group.Compared with the model group,the writhing score decreased,the pain threshold increased,the serum BDNF,SP,and PGF2α levels decreased significantly,the serum PGE2 level increased,and the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus decreased significantly in the preventative moxibustion group and the Western medicine group,while the inter-group differences were significant(P<0.01).Compared with the Western medicine group,the writhing score decreased,the pain threshold increased,the serum BDNF,SP,and PGF2α,levels decreased,the serum PGE2 level increased,and the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus in the preventative moxibustion group decreased significantly,while the inter-group differences were significant(P<0.05 or P<0.01). Conclusion:Preventative moxibustion at Shenque(CV8)and Guanyuan(CV4)can improve the pain sensitization state of rats with dysmenorrhea due to cold-dampness stagnation,down-regulate the mRNA and protein expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus;regulation of the serum SP,PGF2α,and PGE2 levels may be part of the mechanism.

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 investigate the curative effect of Doppler ultrasound-guided management of vascular injury of Gartland type Ⅲ supracondylar fractures of humerus in children.Methods:A prospective study on 18 children with vascular complications of pulseless Gartland type Ⅲ supracondylar humeral fracture who were admitted at Department of Pediatric Orthopedic Surgery, Wuhan Children′s Hospital from March 2017 to March 2018 was conducted.Among these children, 12 were male and 6 were female, with the age of 2.1-8.6 years (mean 4.4 years old), and 10 cases were injured on the left and 8 cases on the right.All patients were satisfied with closed reduction and internal fixation within 24 hours after injury.Before the operation, Doppler ultrasound was performed to determine the shape of brachial artery and it relationship with fracture.During reduction, Doppler ultrasound was used to assess brachial pulse and blood perfusion.Then, the brachial artery was assessed by palpable radial pulse and peripheral blood supply, and elbow joint function was evaluated with Mayo Elbow Performance Score and Flynn criteria.Results:Eighteen patients were followed up for 6 to 12 months (average 9 months) after operation.Before reduction, radial pulse disappeared and peripheral blood supply was good in 18 cases.Preoperative Doppler ultrasonography showed that the brachial artery was located in front of the proximal humerus fracture fragment.A proximal stream of the brachial artery was identified, but the distal blood vessels were compressed by the fragment.A stream of radial artery was identified in 5 patients with Doppler, and no stream was identified in 13 patients.After reduction, the peripheral blood supply of all the affected limbs was good, among which the peripheral blood supply, 1 case returned to normal after 5 minutes waiting.Specifically the palpable radial pulse was immediately restored in 14 patients.The palpable radial pulse did not restore but the peripheral blood supply was good in 4 cases.Intraoperative ultrasonography showed that brachial artery blood flow was good, and palpable radial pulse was restored between 3 to 5 weeks later.In patients with at average sixteen-week neurological follow-up, 5 patients had complete resolution of nerve palsy.No complications such as forearm compartment syndrome and elbow joint dysfunction occurred.Functional outcome as measured by Flynn criteria was excellent in 12 patients, good in 6 patients, the excellent and good rate was 100%.Functional outcome as measured by Mayo Elbow Performance Score was excellent in 17 patients, good in one patient, the excellent and good rate was 100%.Conclusion:Ultrasound-guided is a safe and reliable option to treat pulseless Gartland type Ⅲ supracondylar humeral fracture.