ObjectiveTo explore the effect of external fixation of damage control operation for treating severe multiple injury patients complicated with open tibia and fibula fractures.MethodsFiftyeight severe multiple injury patients complicated with open tibia and fibula fractures from June 2006 to February 2011 were reviewed.According to the control strategy for tibia and fibula fractures,all patients were divided into two groups:32 patients in observation group were treated with external fixation and 26 patients in control group were treated with calcaneal traction or gypsum external fixation.Wound healing time,fracture healing time,and the incidence rate of acute respiratory distress syndrome (ARDS) and multiorgan dysfunction syndrome (MODS),mortality and wound infection rate within 1 month between two groups were compared.ResultsIn observation group,incidence of ARDS and wound infection rate within 1 month,wound healing time was 6.25％(2/32),31.25％(10/32) and (21.47 ± 6.84) d,respectively,while 24.00％(6/25),64.00％ ( 16/25 ) and (30.26 ± 2.18 ) d in control group,respectively,there were significant differences between two groups(P ＜0.05).The differences of the incidence rate of MODS and mortality within 1 month,fracture healing time of observation group were not statistically significant compared with control group (P ＞ 0.05).ConclusionExternal fixation of damage control operation for treating severe multiple injury patients complicated with open tibia and fibula fractures has the advantages of reducing complication occurrence and improving success rate.

Objective To evaluate the clinical significance of serum procalcitonin (PCT)in early diagnosis of pul-monary tuberculosis (PTB)complicated with pulmonary infection.Methods Clinical data of active PTB patients admitted to a hospital between August and December 2013 were collected,patients were divided into bacterial infec-tion group(n=104),fungal infection group(n=37)and control group (n=95)according to whether patients were associated with bacterial infection,fungal infection,and without infection,serum PCT concentrations in three groups were compared,receiver operating characteristic (ROC)curve analysis was conducted.Results The median PCT concentrations in bacterial infection and fungal infection group was 0.44ng/mL and 0.30ng/mL respectively, which was significantly higher than 0.16ng/mL of control group(Z =9.49,3.51 respectively,both P <0.001 ).The area under curve (AUC)was 0.89(0.84-0.93)and 0.69(0.61 -0.77)respectively;cut-off point was 0.31 ng/mL and 0.27 ng/mL respectively;sensitivity was 79.81%(70.57%-86.80%)and 59.46%(42.19%-74.80%)respectively;specificity was 83.16%(73.79%-89.78%)and 73.68%(63.48%-81.95%)respectively.Conclusion PCT level is a valuable predictor for early diagnosis of PTB complicated with pulmonary infection,and can provide reference for the rational use of antimicrobial agents.

Objective To explore the clinical application effect of squamous cell carcinoma antigen (SCC) ,cancer embryo antigen (CEA) and carbohydrate antigen 125(CA125) chemiluminescence combined detection in early diagnosis of lung cancer .Methods The SCC ,CEA and CA125 levels in the patients with early stage lung cancer ,benign lung diseases and healthy subjects undergoing physical examination were detected by using the automatic chemiluminescence immune analyzer .Results The concentrations of three tumor markers in the lung cancer group were significantly higher than those in the benign disease group and control group (P<0 .05);t he concentrations of CEA and CA125 in the adenocarcinoma patients with were significantly higher than those in the patients with squamous cell carcinoma and small cell carcinoma (P<0 .05) .The concentration of SCC in the patients with squamous cell carcinoma was significantly higher than that in the patients with adenocarcinoma and small cell lung cancer (P<0 .05) .The sen-sitivity ,specificity ,positive predictive value and negative predictive value of combined detection of three tumor markers were signifi-cantly better than that of single tumor marker ,and the difference was statistically significant (P<0 .05) .Conclusion The combined detection of SCC ,CEA and CA125 can be used in the early diagnosis of lung cancer and has the promotion value .

Objective To explore whether 5 to -7-day of folic acid supplementation prior to pemetrexed therapy is needed. Methods We retrospectively evaluate the outcomes of non-small cell lung cancer patients received less than the advised folic acid premedication. Seventy patients with Ⅲ-Ⅳ NSCLC were randomly divided into two groups: patients who initiated vitamin supplementation on 7 days before the first dose of pemetrexed (group A) and patients who initiated vitamin supplementation on the day of the first dose of pemetrexed (group B). Results In group A and group B, CR 0 and 0, PR 10 and 8, the response rates of 28.6% and 22.9% were observed, respectively. There was no statistically significant differences between these two groups. No significant differences were observed in the incidence of hematologic and non-hematologic toxicities. Conclusions The initiation of pemetrexed-based chemotherapy does not need to accommodate a vitamin supplementation schedule.

Objective To analyze the relationship between the expression of protection of telomeres 1 (POT1) and the pathogenesis of acute myeloid leukemia (AML). Methods 62 patients with de novo AML (case group) and 10 patients with iron deficiency anemia (control group) were enrolled in this study. The quantitative real-time polymerase chain reaction (PCR) and Western blot were used to detect the expression of POT1 in AML patients. Results There were 62 de novo AML patients, including 2 cases M1, 14 cases M2, 12 cases M3, 14 cases M4, 17 cases M5, 2 cases M6 and 1 case AML without classification. According to the risk stratification, high risk group (24 cases), medium risk group (22 cases) and low risk group (16 cases) were divided. Compared with that in the controls, POT1 expression levels in patients with AML were significantly decreased both in mRNA and protein level (P< 0.05). The relative expression levels of POT1 mRNA and protein in patients with M2, M4 and M5 were significantly lower than those in the controls (P< 0.05). The expression levels of POT1 in high risk group, medium risk group and low risk group were significantly decreased than those in the controls (P<0.05). Compared with that in the controls, The relative POT1 mRNA expression was significantly decreased in M3 patients (P< 0.05), but not in protein level. POT1 protein expression was showed both in the cytoplasm and nucleus. There was no significant difference of the expression of POT1 protein between cytoplasm and nucleus (P> 0.05). Conclusions POT1 may be involved in the pathogenesis of AML. POT1 protein expresses in both cytoplasm and nucleus, and the regulatory mechanism may be related to the telomere length.

BACKGROUND:The dynamic changes of hypoxia inducible factor-1 alpha (HIF-α) and inducible nitric oxide synthase (iNOS) genes in the pulmonary artery wall during the process of hypoxic pulmonary hypertension (HPH) development need to be investigated.OBJECTIVE: This study was to observe the gene expressions of HIF-α and iNOS in the pulmonary artery wall at the different hypoxic time points, and to investigate their effects in HPH development.DESIGN: Controlled observation animal experiment.SETTING: Department of Respiration, Second Hospital Affiliated to Nanhua University.MATERIALS: Forty healthy male Wistar rats of clean grade, aged 6-8 weeks, with body mass of (220±10) g were involved in this study. They were randomized into control group (n =8) and hypoxia group (n =32). Four time points, i.e.3, 7, 14, and 21 days after hypoxia were set for the animals in the hypoxia group, 8 rats at each time point.METHODS: This study was carried out in the Institute of Oncology, Nanhua University between August 2004 and December 2005. Rats in the hypoixa group were treated according to the method reported by Li et al. Hypoxia treatment was omitted for the rats in the control group. At each time point, the rats were anesthetized, and then a micro-catheter was inserted into the right jugular vein and connected to a multichannel physiologic recorder, which was used for detecting the mean pulmonary arterial pressure (mPAP). The heart of each euthanized rat was taken out, and its right ventricle (RV), and left ventricle and septum (LV+S) were weighted. Right ventrical hypertrophy index (RVHI) reflected right ventricle hypertrophy degree. Right upper lung tissue of rat was harvested for haematoxylin & eosin (HE) staining and elastic fiber staining. Pathological image analysis software was used to determine pulmonary arterial wall area/total vascular area, lumina area/total vascular area, smooth muscle cell density in the media of pulmonary arteriole, and media thickness of pulmonary arteriole, which were used as remodeling indexes of pulmonary arteriole. HIF-1α and iNOS in the pulmonary arteriole were performed in situ hybridization and immunohistochemical detection. The mean absorbance of pulmonary arteriole wall was used as the relative content of mRNA expression and protein level.MAIN OUTCOME MEASURES: ①Changes in mPAP, RV hypertrophy degree and remodeling indexes of pulmonary arteriole of rats. ② Expressions of HIF-α and iNOS as well as their correlations with mPAP and remodeling indexes of pulmonary arteriole.RESULTS: All the 40 Wistar rats were involved in the final analysis. ①At hypoxia 7 days, mPAP was significantly higher than that of control group (P ＜ 0.05). mPAP reached to the high level at hypoxia 14 days, and then maintained at this level. ② At hypoxia 14 days, RVHI was higher than that of control group (P ＜ 0.05). ③ At hypoxia 7 days, pulmonary arteriole wall was thickened, and lumina became narrowed. There were significant differences in pulmonary arterial wall area/total vascular area, lumina area/total vascular area between hypoxia group and control group (P ＜ 0.05). At hypoxia 14 days, smooth muscle cell density in the intima-media of pulmonary arteriole, and intima-media thickness of pulmonary arteriole were significantly increased (P ＜ 0.05). At hypoxia 21 days, lumina was further narrowed, and obvious smooth muscle hyperplasy was found. HIF-1α and iNOS mRNA expression presented weak-positive in the control group; The relative content of HIF-1α mRNA did not significantly alter at hypoxia 3 and 7 days, but was obviously increased at hypoxia 14 days ,and after this, it maintained at high level. iNOS mRNA was markedly higher than that in the control group at hypoxia 3 days, reached its peak at hypoxia 7 days, was close to the level in the control group at hypoxia 14 days, and enhanced again at hypoxia 21 days, but it was still lower than that at hypoxia 3 days. ⑤HIF-α was mainly found in the intima and media, while iNOS was found in the whole layer of vessels. iNOS was weakly expressed in the intima and media of pulmonary vessels in the control group. No obvious difference in iNOS existed at hypoxia 3 days as compared with control group. iNOS was obviously expressed in media and intima at hypoxia 7 days. Vascular media was thickened at hypoxia 14 days, and the expression of iNOS was enhanced. iNOS was not found in the vascular adventitia in the control group, but was found in the vascular adventitia in the hypoxia group. ⑥mPAP was positively correlated with pulmonary vascular remodeling (r =0.976, P ＜ 0.01 ), and HIF-1α mRNA was positively correlated with iNOS protein(r =0.927, P ＜ 0.05).CONCLUSION: Both HIF-1α and iNOS exert effects in the process of HPH development of rats, and HIF-1α and iNOS gene expression may be mutually regulated.

BACKGROUND: How to reconstruct tissue-engineered bone with structure similar to natural bone iS a problem in the development of tissue engineering. Cell sheet engineering technology enables novel approaches to construction of tissue-engineered bone. OBJECTIVE: To observe the biocompatibility of call sheets to decalcified bone matrix (DBM) and their growth on DBM. DESIGN, TIME AND SETTING: An in vitro observation was performed at the Central Laboratory, Affiliated Hospital, Qingdao University Medical College between June and September 2009.MATERIALS: Dog bone marrow stromal cell sheets were prepared using temperatura-responsive medium. Dog DBM was prepared by defatting, decalcification, and noncotlagen protein removal procedures. METHODS: DBM surface was covered by call sheets prepared by temperature-responsive technology and cultured with DMEM containing 10% fetal bovine serum and osteoinductive agent.MAIN OUTCOME MEASURES: Under scanning electron microscope, DBM structure, as well as the attachment and growth of cell sheets on DBM surface, was observed. Porosity and aperture size of DBM were calculated. RESULTS: DBM exhibited a three-dimensional latticed structure, with a porosity of approximately 75%. The mean aperture size was (250.11±98.89) μm, exhibiting a normal distribution. Cell sheets well attached to and grew on DBM surface, and rapidly proliferated.CONCLUSION: Cell sheets show good biocompatibility to DBM. DBM/cell sheets complex can be applied in tissue-engineered bones, which promotes the construction of tissue-engineered bone with structure similar to natural bone.

BACKGROUND:There are some disadvantages in harvesting and transferring cells in the traditional tissue engineering technique,and it is difficult to form dense tissues,which significantly limits the development of tissue engineering.OBJECTIVE:To explore the culture and fabrication of dog bone marrow mesenchymal stem cell(BMSC)sheet in vitro.METHODS:Bone marrow was extracted from dogs following anesthesia.BMSCs were isolated with the method of density gradient centrifugation in vitro.BMSCs at passage 4 at a density of 1×10~9/L were incubated in the temperature-responsive culture dishes with a diameter of 3.5 cm,and cultured in an incubator at 37 ℃,5% CO_2 and saturated humidity.The temperature of the incubator was changed from to 37 V to 20 ℃ to prepare BMSCs cell sheet for 20 minutes.Cell morphological changes and cell sheet formation were observed under an inverted microscope.RESULTS AND CONCLUSION:Dog BMSCs following 24 hours of primary culture presented ellipse or polygonal shape.Most cells adhered at hour 72,and cell colonies were visible at day 7.Cells showed long spindle and completely confluence at day 12,with unclear boundary.BMSCs in the temperature-responsive culture dishes presented short spindle shape,and gradually separated from the dish bottom,forming entire cell sheet containing extracellular matrix at 20 V.These verified that dog BMSCs can be effectively obtained with method of density gradient centrifugation.Complete cell sheet layer can be fabricated with temperature-responsive culture dishes.

Objective To investigate the effect of micro-ecological agent on the intestinal flora of patients with posterior lumbar spine surgery.Methods Seventy-two patients with lumbar spine surgery were selected as our subjects.Of them,42 patients with occurred the intestinal flora dysfunction were served as experimental group (group A),other 30 patients were as control group (group B).The levels of plasma endotoxin,plasma tumor necrosis factor-α (TNF-α),Interleukin 6 (IL-6) were detected before and after operation.The patients in group A were treated with micro-ecological agent Jinshuangqi.Collected stool samples at preoperative,postoperative first natural defecation (after 1-4 d),and compared intestinal flora difference and Bifidobacterium (B)/Enterobacteriaceae (E) value of two groups patients.Results The levels of plasma endotoxin in group A were (1.82 ±0.12),(2.29 ±0.15),(1.91 ±0.08) ng/L at before surgery,the 2nd and 7th day after surgery,and that in group B were(1.91 ±0.21),(2.35 ±0.16),(2.26 ±0.24) ng/L The TNF-α level in group A were (275 ±51),(309 ±45),(276 ±34) ng/L and that in group B were (269 ±48),(318±67),(297 ±53) ng/L at before surgery,the 2nd and 7th day after surgery.The IL-6 levels in group A were (138 ± 22),(159 ± 15),(137 ± 17) ng/L and that in group B were (159 ± 16),(187 ± 19),(174 ± 21) ng/Lat before surgery,the 2nd and 7th day after surgery.The repeated measured repeated measures analysis of variance showed that among three indexes of plasma endotoxin,TNF-α,IL-6 levels,differences of between group and interaction were statistically significant (P ＜ 0.05),but there was no significant difference within groups (P＞ 0.05).The levels of plasma endotoxin,TNF-α,IL-6 decreased after surgery ware gradually decreasing with hospitalization prolonged.There was significant difference between before surgery and at the 2nd,7th day after surgery(P ＜ 0.05),and there was significant difference between the 2nd and the 7th day after surgery(P＜ 0.05).At the 2nd day after surgery,there was significant difference between group A and group B in terms of the level of plasma endotoxin,TNF-α,IL-6.At the 7th day,the levels of plasma endotoxin,TNF-α,IL-6 were slowly decreased in group A,and there were significant difference compared to group B (P ＜ 0.05).The numbers of intestinal Bifidobacterium,Bacteroides,and B/E values of group A were decreased after surgery compared to group B((9.17 ±0.54) lg cfu/g vs.(10.01 ± 0.75) lg cfu/g,(9.23 ± 0.47) lg cfu/g vs.(10.09 ± 0.81)lg cfu/g,(1.01 ± 0.16) vs.(1.20 ± 0.11)),and the difference was significant (P =0.031,0.042,0.029respectively).The levels of Enterobacter and Enterococcus in group A were (9.11 ± 1.02) lg cfu/g,(7.80± 1.02) lg cfu/g,higher than that of group B ((8.81 ± 0.89) lg cfu/g,(7.29 ± 0.98) lg cfu/g(P =0.037,0.043)).There was no significant difference between two groups in terms of small clostridium and lactobacillus (P ＞ 0.05).After treated with micro-ecological agent Jinshuangqi,the levels of intestinal Bifidobacterium,Bacteroides,and B/E value at was increased compared to before treated in group A (Bifidobacterium:(13.01±0.87) lg cfu/g vs.(9.17 ±0.54) lg cfu/g; Bacteroides:(14.12 ±0.75) lg cfu/g vs.(9.23 ±0.47)lg cfu/g; B/E value:(1.28 ± 0.45) vs.(1.01 ± 0.16) ;P =0.045,0.034,0.038 respectively).No significant difference was seen in terms of the levels of Enterobacter,Enterococcus,small clostridium and lactobacillus between two groups(8.71 ±0.91) lg cfu/g vs.(9.11 ± 1.02) lg cfu/g,(7.01 ±0.54) lg cfu/g vs.(7.80± 1.02) lg cfu/g,(5.23 ± 1.04) lg cfu/g vs.(5.15 ± 0.89) lg cfu/g,(6.71 ± 1.04) lg cfu/g vs.(6.53± 0.86) lg cfu/g respectively; P ＞ 0.05).Conclusion Posterior lumbar operation patients with intestinal dysbacteriosis often associated with endotoxemia and inflammatory reaction,the levels of bifidobacteria,Bacteroides intestinal Enterobacteriaceae decrease while Enterococcus,opportunistic pathogens increase,and intestinal microbial colonization resistance decrease.Micro-ecological agent Jinshuangqi treatment can ease the body's endotoxemia and inflammatory response,improved lumbar surgery intestinal flora after surgery,and beneficial to rebuild the intestinal microflora balance.

Objective To investigate the relationship between genetic variants in the Toll-like receptor (TLR) pathway genes and susceptibility of gastric cancer (GC) and esophageal squamous cell carcinoma (ESCC).Methods The data of whole genome association studies of the high-risk population of GC and ESCC in China were analyzed by adaptive rank-truncated product (ARTP) method in pathway and gene level.The associations between single nucleotide polymorphism (SNP) and susceptibility of GC and ESCC were analyzed with additive model of unconditional Logistic regressions.PLINK 1.07 and SPSS 19.0 software were performed for statistical analyses,and ARTP package in R3.0.2 was used for pathway and gene level analysis.Results In gene-level analyses,eight genes were found to be associated with susceptibility of GC (P ＜0.05) and six genes were associated with susceptibility of ESCC (P ＜ 0.05).In single SNP-level analyses,21 SNPs were statistically correlated with susceptibility of GC (P ＜ 0.01),and 11 SNPs were statistically correlated with susceptibility of ESCC (P ＜0.01).Conclusions Some genetic variants in TLR pathway are associated with risk of GC and ESCC.The potential molecular mechanisms need further investigation.