Objective To investigate the relationship between the expression of Toll-like receptor 4 (TLR4) on mononuclear cells in peripheral blood and changes of serum vitamin D level in children with bronchiolitis.Methods The children who were diagnosed as bronchiolitis and received treatment in the Second Affiliated Hospital of Harbin Medical University from October 2013 to January 2014 were chosen as the pre-treatment group,and then divided them into moderate group and severe group according to the clinical symptoms,20 cases for each group.Then the cases in pre-treatment group who recovered after treatment were recruited as the after-treatment group,and the children who were healthy and medical examination in the Second Affiliated Hospital of Harbin Medical University in the same period were recruited as the healthy control group.The expressions of TLR4 on CD14 labeled mononuclear cells in the periphera were measured by flow cytometry.The level of 25 (OH) D in serum was detected by enzyme linked immunosorbent assay (ELISA).Results (1) The expression level in children with bronchiolitis of TLR4:the mode-rate group [(18.98 ±2.29)％] and severe group [(30.13 ±2.13)％] increased significantly (P ＜0.05) compared with control group [(1.17 ± 0.57) ％].And the expression level of moderate group [(2.02 ± 0.48) ％] and severe group [(11.43 ± 1.52) ％] decreased significantly after treatment (P ＜0.05).(2) Serum vitamin D level in children with bronchiolitis of the moderate group[(17.16 ± 3.34) μg/L] and severe group [(6.56 ± 2.28) μg/L] were lower than healthy control group [(53.69 ± 20.18) μg/L] before treatment (P ＜ 0.05),especially the severe group [(6.56 ±2.28) μg/L].The level of moderate group [(9.59 ± 2.31) μg/L] and severe group [(4.70 ± 0.67) μg/L] became lower after treatment (P ＜ 0.05).(3) Both severe group (r =-0.491,P ＜ 0.05) before treatment and moderate group (r =-0.436,P ＜ 0.05) after treatment showed negative correlation between TLR4 on mononuclear cells in peripheral blood and serum 25 (OH)D level in children with bronchiolitis.And no correlation was found among healthy control group,moderate group before treatment and severe group after treatment (P ＞ 0.05).Conclusions The conditions of children with bronchiolitis was positively correlated with the expression level of TLR4,and negatively correlated with the vitamin D level.The serum 25 (OH) D decreased steadily during the treatment.The expression of TLR4 in monocytes has a certain correlation with the level of vitamin D in children with bronchiolitis.

BACKGROUND:There remains controversy that whether patel ar resurfacing in total knee arthroplasty and whether patel ar chondromalacia has influence on clinical outcomes.
OBJECTIVE:To evaluate anterior knee pain, clinical function and effects of patel ar chondromalacia on clinical outcomes after total knee arthroplasty without patel ar resurfacing for degenerative osteoarthritis.
METHODS:Clinical data of 162 patients (162 knees) with degenerative osteoarthritis undergoing total knee arthroplasty without patel ar resurfacing from June 2008 to February 2010 were retrospectively analyzed. Outerbridge classification was used for patel ar chondromalacia, Visual Analogue Scale for anterior knee pain, and the Knee Society clinical scoring system for clinical function.
RESULTS AND CONCLUSION:The incision of al patients reached stage-I healing. The patel ar chondromalacia:grade I in 18 patients, grade II in 36 patients, grade III in 62 patients, and grade IV in 35 patients. At the final fol ow-up, there were six (4.0%) patients with anterior knee pain, including four cases of mild pain and two cases of moderate pain, no severe pain. The mean Knee Society clinical scoring system scores and patel ar score were obviously elevated. Outerbridge classification did not affect the incidence of anterior knee pain after replacement (χ2=0.42, P=0.94), the Knee Society clinical scoring system score (knee score:F=1.83, P=0.14;functional score:F=0.56, P=0.64) and partel ar score (F=0.78, P=0.51). These data suggested that total knee arthroplasty without patel ar resurfacing for degenerative osteoarthritis can obtain satisfactory clinical outcomes, and the patel ar chondromalacia may not affect the clinical outcomes.

Objective To investigate the effect of dexmedetomidine (DEX) on inflammatory mediators in different periods of sepsis and its impact on survival rate of rats with sepsis.Methods Ninety male Wistar rats were randomly divided into three groups:control group (group C),sepsis model group (group S),and DEX treatment group (group D).Lipopolysaccharide (LPS) was injected to the vein to reproduce a classic sepsis model,while in control group,equivalent volume of NS was used instead.Different treatments were given to the corresponding groups,and then venous blood was taken at given intervals.Behavioral changes and mortality rate of rats within 48 hours after experiment were recorded.The levels of the early inflammatory mediators such as tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6) and the late inflammatory mediators such as high-mobility group boxl protein (HMGB1) in blood serum were detected by ELISA.Results Among three groups,there were significant differences in mortality rate of rats,the serum level of TNF-αt,IL-6 and HMGB1 (all P ＜ 0.05).Conclusion DEX has a certain anti-inflammatory effects on early and late stages of sepsis in rats with sepsis,and can also reduce the mortality rate.

Silkworm is a holometabolous insect of Lepidoptera. During metamorphosis, significant morphological changes happen including the dissociation of old tissues and remodeling of new tissues. It has been reported that cathepsins are involved in these processes. Cathepsin is a kind of intracellular proteinase that exists in many species. It includes some subfamilies like cathepsin B, H and L. The studies on cathepsin are useful for clarifying the details of silkworm metamorphosis process. In total, 13 cathepsins were identified by screening the silkworm genome database. The basic information and the expression patterns about these genes were analyzed. Interestingly, an ovary-specific cathepsin L gene (Gene ID: BGIBMGAOO4622) was investigated by the data of silkworm microarray and real-time quantitative PCR (qPCR). The full-length cDNA is 1,209 bp, encoding a protein with 402 amino acids. Sequences alignment revealed that it has a high sequence similarity with cathepsin L of other species, and it is highly conserved in the active-site of the enzyme. The phylogenetic analysis showed that ovary-specific cathepsin L is clustered with other lepidopterous insects. Furthermore, this gene was cloned and prokaryotic expressed. Recombinant protein was present in inclusion body. Importantly, the qPCR result showed that the expression level of this gene is increasing during the early stage of pupal development and reaches the highest value at the 3rd day of pupal stage, which suggests that this gene may be involved in the process of development of the ovary and oocyte.
Animals ; Bombyx ; genetics ; Cathepsins ; genetics ; Insect Proteins ; genetics ; Phylogeny

Objective To investigate the effect of decompression with long-segment (L) or short-segment (S) fusion on the outcomes of the surgical treatment for degenerative adult scoliosis (ADS) patiens.Methods A retrospective study on 32 patients treated in our department for ADS from April 2013 to May 2015 was carried out, including 12 male and 20 female (1 : 1.7).Their average age was 66.4 (range: 51-77 years).All patients underwent decompression and fusion surgeries through posterior approach.They were divided into long-segment fusion group (L) and short-segment fusion group (S) according to fusion range.During follow-ups (FU), clinical outcomes were assessed by means of visual analog scale (VAS) and Oswestry disability index (ODI).Radiographic evaluation on full-length standing film included coronal Cobb's angle, distant between C7plumb line and center sacral vertical line (C7PL-CSVL), thoracic kyphosis (TK) angle, lumbar lordosis (LL) angle, pelvic incidence (PI), and pelvic tilt (PT), PI-LL, sagittal vertical axis(SVA).Postoperative complications were also recorded.Results All patients were followed up for average 2.2 years (range:1.5-3.5 years).No significant difference of age or gender was found between two groups (L: 22, S:10) of patients (P=0.066, 0.182).As for the fusion segments, group L (6.3±1.5) was more than group S (2.9±0.3) (P=0.001).Operation time and blood loss of group L were statistically more than group S (P＜0.05).Postoperative VAS sores of back pain and leg pain as well as ODI were all improved significantly in two groups (P＜0.05).At the final FU, back pain VAS was more in group L than in group S (P＜0.05) , but no significant difference was found in leg pain VAS between two groups (P＞0.05);at the final FU, group L's ODI showed better functional recovery than group S's[(12.8±9.3)％ vs.(25.4±11.4)％, P＜0.05].With no obvious difference in the two groups (P＞0.05), coronal Cobb's angle corrected more satisfactional in group L than group S (P＜0.05).The same situation was found in C7-CSVL correction in two goup in FU (P＜0.05).Sagital balance was restored to normal alignment better in group L than group S, with increase of lumbar lordosis after surgery.The overall incidence of postoperative complications was 31.3％, including wound infection, cerebrospinal fluid leakage, transient neurological symptoms and internal fixed rod breakage, more common in group L than group S.Conclusions Decompression and fusion with internal fixation showed good clinical outcomes in the treatment of ADS.Long-segment fusion yielded better coronal and sagittal correction outcomes with higher peri-operation risks;however, short-segment fusion showed higher safety with relatively inferior correction effect.Appropriate fusion mode should be chose according to the patient's deformity features.

Objective To investigate the effect of volume therapy with 6％ hydroxyethyl starch (HES) 130/0.4 on mesenteric microcirculation in a rabbit model of hypovolemia.Methods Sixty-four adult male rabbits,weighing 2.0-2.3 kg,were randomly divided into 4 groups( n =16 each):control group (group C),hypovolemia group (group HM),Ringer's solution group (group RS) and 6％ HES 130/0.4 group (group HES).The animals were anesthetized with sodiun pentobarbital 3.0-3.5 mg/kg.Femoral artery,femoral vein and right internal jugular vein were cannulated for MAP monitoring,blood withdrawing and fluid adminstration.Hypovolemia was induced by withdrawing blood from femoral vein of 30％ blood volume in 30 min in groups HM,RS and HES.In group C,no blood was withdrawed.In group RS,Ringer's solution 3 times of the volume of blood withdrawn was infused,while 6％ HES 130/0.4 equal volume of blood withdrawn was infused via right internal jugular vein immediately at the end of blood withdrawing.MAP and HR were recorded and femoral artery and femoral vein blood samples were taken before blood withdrawing (T0),immediately at the end of blood withdrawing(T1 ),immediately at the end of volume therapy(T2 ) and 30 min after volume therapy(T3 ).DO2,VO2 and ERO2 were calculated.Diameter and blood flow speed of microvessel were determined.Results Compared with group C,HR was significantly increased,MAP decreased,diameter and blood flow speed of microvessel were decreased at T1～3,DO2 was increased　at T1 in group HM.Compared with group HM,MAP was significantly increased at T2,HR decreased at T2,3,DO2and VO2 were increased at T1～3,diameter of arteriole was increased at T2 whlie decreased at T3,diameter of veinule　and blood flow speed of microvessel were increased at T2,3 in group RS,MAP was significantly increased at T2,HR　was decreased whlie DO2 and VO2 were decreased,VO2,ERO2,diameter and blood flow speed of microvessel were　increased at T2,3 in group HES.Compared with group RS,DO2,VO2 and ERO2 were significantly decreased at T3　in group HES.Conclusion Volume therapy with 6％ HES 130/0.4 can improve mesenteric microcirculation,organ perfusion and oxygen metabolic in a rabbit model of hypovolemia.

This study aimed to develop the calculation method of pre-hospital emergency physician allocation gap and apply it to Shanghai.In order to reduce the ambulance dispatch lag frequency, through the analysis of its da-ta in the Shanghai urban area, the research group obtained the gap and extended the data to Shanghai city.The peak method establishes the association between pre-hospital emergency physician increment and the ambulance dispatch lag decrement.Based on descriptive statistics, the peak method by which the Shanghai ambulance dispatch lag data were analyzed uses the SAS programming software.This method of using programming software provides it with good reliability and validity.After an increase of 40 duty vehicles (381 pre-hospital emergency physicians), the ambu-lance dispatch lag ratio would drop from 25.61 percent to 0.22.Therefore, the association between the pre-hospital emergency physician increment and the ambulance dispatch lag decrement was established and can provide a scientif-ic evidence for the policy formulation.

Objective:The research aimed to figure out the geographic availability of the residence in the aspect of receiving health service thereby studying the spatial distribution regulation and the distance attenuation coefficient of the inpatients.It also provides the decision-making basis for the regional health planning.Methods: Select 23 medical institutions and randomly pick up patients from each of them, calculate the distance between the inpatients home and the medical institutions, then simulate the corresponding probability patients using the Geographical Infor-mation System ( GIS) .Perform a variety of curve fitting models according to the determination and simulation graph-ics coefficient choosing the most appropriate model, and draw corresponding the distance attenuation coefficient.Re-sults:An exponential curve model is better than power curve model for the secondary medical institutions inpatients distribution and contrarily for those in the tertiary medical institutions.The mean distance attenuation coefficient was 0.51 for exponential curve model of secondary comprehensive medical institutions inpatients and its average was 0.50 for the secondary specialist medical institutions inpatients.The mean distance attenuation coefficient was 0.84 for the power curve model of the tertiary comprehensive medical institutions inpatient and its average was 0.53 for tertiary specialist medical institutions inpatients.Conclusion: The secondary medical institutions inpatient probable attenua-tion trends with distance are faster than in tertiary medical institutions, those for the comprehensive medical institu-tions inpatient are faster than in the specialist medical institutions.In the regional health planning, for different lev-els, categories of medical institutions should adopt different distance attenuation coefficients to calculate the health re-sources level.

Objective To analyze the variations of glycerol-3-phosphate dehydrogenase 1 like gene (GPD1-L) and address the association with sudden m anhood death syndrom e (SMDS). Methods The genom ic DNA was extracted from blood sam ples of the SMDS group and the norm alcontrolgroup.The exons, exon-in-tron boundaries and 3′-U TRs of coding region of GPD1-L w ere PCRam plified and DNAsequenced di-rectly to confirm the types of variations. The genotype frequency and allele frequency w ere analyzed statistically. Results There w ere tw ovariants in the SMDS group, c.465C>Tand c.*18G>T, the latter existed certain degree difference of genotype distribution and allele frequency betw een the SMDS group and the control group, but there was no statistically significant (P>0.05). Conclusion The relation be-tw een gene m utation of GPD1-L and the occurrence of Chinese SMDS deserves a further research.

We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis,expressing P61 protein with biological activity in E.coli,and lay a foundation for further studies related to P61.P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and induced with IPTG.The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit.The protein of P61was successfully expressed in E.coli with solubility and high catalase activity,and could be identified by anti-N.brasiliensis sera from mice.The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.