BACKGROUND:Orthopedists should pay more attentions to nonunion prevention in view of nonunion treatment, that is, active interventions should be taken to avoid bone delayed union and nonunion.
OBJECTIVE:To explore the effect of composite tissue-engineered scaffold constructed by nano-hydroxyapatite/polyamide 66 (nHA/PA 66) combined with bone marrow mesenchymal stem cels to repair a femoral fracture with severe nonunion.
METHODS:Rat bone marrow mesenchymal stem cels were isolated and culturedin vitro, and then they were divided into three groups: bone marrow mesenchymal stem cels without osteogenic induction, with osteogenic induction or combined with nHA/PA 66 folowed by osteogenic induction as control group, test group or composite group, respectively. Then osteogenic differentiation of bone marrow mesenchymal stem celsin vitrowas analyzed by measuring alkaline phosphatase activity and alizarin red staining, cel adhesion on the nHA/PA 66 was observed using scanning electron microscopy, and the celgrowth and proliferation were detected by MTT assay. In the meanwhile, established Sprague-Dawley rat femur nonunion models were randomly divided into three groups: the areas of nonunion were implanted with nothing as blank control group,those were with nHA/PA 66 as simple scaffold group, and the others were with nHA/PA 66 combined with bone marrow mesenchymal stem cels as composite scaffold group. Afterwards, X-ray examination, micro-CT and Masson staining were used to evaluate the femoral healing.
RESULTS AND CONCLUSION:At 6 and 12 days after osteogenic induction, alkaline phosphatase activity in the test group was significantly higher than that in the control group; at 14 days, compared with the control group, the amount of mineralized nodules in the test group was significantly higher, which indicated that bone marrow mesenchymal stem cels after osteogenic induction could differentiate into osteoblasts. Attached cels spread wel on the scaffold with good proliferation activity, suggesting that nHA/PA 66 is suitable for cel adherence, proliferation and osteogenic differentiation. Besides, at 12 weeks after modeling, in the blank control group, no calus appeared in the nonunion region. In the simple scaffold group, the broken femur did not heal at 8 and 12 weeks after surgery. In the composite scaffold group, the broken femur did not heal at 8 weeks, but a lot of calus appeared; at 12 weeks, bone healing achieved and the scaffold was encased and absorbed.These findings demonstrate that the tissue-engineered bone scaffolds constructed by bone marrow mesenchymal stem cels and nHA/PA 66 effectively prevent bone nonunion by accelerating femoral healing in a rat femur nonunion model.

Objective To explore the biocompatibility of nano-hydroxyapatite/polyamide 66 (nHA/PA66) with human bone mesenchymal stem cells (hBMSCs) after osteogenic induction.Methods After hBMSCs were isolated and cultured in vitro,the experiment was conducted in 3 groups.Group A were hBMSCs subjected to no osteogenic induction,group B hBMSCs subjected to osteogenic induction,and group C was the composite of nHA/PA66 with hBMSCs subjected to osteogenic induction.Adhesion of the cells onto the nHA/PA66 in group C was observed by electron microscope scanning.Growth and proliferation of the cells in groups B and C were detected by MTI test at 1,2 and 3 weeks.The ability of osteogenic differentiation of hBMSCs in vitro was analyzed by alkaline phosphatase (ALP) activity and alizarin red staining.The ability of osteogenic differentiation of hBMSCs on nHA/PA66 was tested by ALP activity.Results Electron microscope scanning showed that the cells spread and attached well on the surface of the composite scaffold in group C;the proliferation of the cells in groups B and C showed no significant difference (P ＞ O.05).These suggested that the proliferation of hBMSCs was not affected by nHA/PA66.The number of mineralized nodules in group B was significantly larger than in group A (P ＜ O.05);the ALP activity of the cells in group A was significantly lower than in group B at 6 and 12 days (P ＜ 0.05);no significant differences were observed between groups B and C (P ＞ 0.05).These indicated that the hBMSCs were capable of osteogenic differentiation which was not affected by nHA/PA66.In groups B and C,the ALP activity of the cells at 12 days was significantly higher than at 6 days,indicating the ALP activity increased with increased induction time (P ＜ 0.05).Conclusion nHA/PA66 can be used as a carrier of hBMSCs in bone tissue engineering because hBMSCs can well adhere to,proliferate,and differentiate into bone on nHA/PA66 scaffolds.

Objective To assess the safety and effectiveness of artificial ascites assisted thermal ablation for hepatic tumors adjacent to the gastrointestinal tract in patients with a history of abdominal surgery . Methods Thirty‐two patients (33 lesions located adjacent to the gastrointestinal tract) with a history of abdominal surgery were included in the study . Method ① :normal saline was injected into abdominal cavity to form water insulation band between liver and gastrointestinal tract . Method② :normal saline was injected with appropriate pressure to form local water insulation band between the liver and gastrointestinal tract . Method③ :normal saline was injected continuously at the interval between liver and gastrointestinal tract to flush away heat energy caused by ablation . All the patients were checked for gastrointestinal tract injury after ablation . During one month after ablation ,CT /MR was performed to evaluate whether the lesions were completely ablated . Results Respectively ,the numbers of lesion received method ① ,② and ③ were 27(81 .8% ) ,4(12 .1% ) and 2(6 .1% ) ,while the usage of normal saline were 400~2 000 ml ,600~800 ml and 1 000~1 500 ml . No gastrointestinal tract injury occurred . CT/MR scan during one month after ablation showed that all the 33 lesions had been completely ablated . Conclusions In patients with a history of abdominal surgery ,artificial ascites is a safe and effective method in assistance of thermal ablation for hepatic tumors adjacent to the gastrointestinal tract .

Objective To determine the level of soluble B7-H1 (sB7-H1) in serum of patients with colorectal cancer ( CRC) , and to investigate its clinical application value in CRC .Methods 152 cases of CRC, 57 cases of benign colorectal diseases and 59 healthy subjects were enrolled .ELISA was used to determine the sB7-H1 level in serum.The levels of carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9) and carbohydrate antigen 72-4 (CA72-4) were determined by electrochemiluminescence. The level of carbohydrate antigen 50 (CA-50) was determined by chemiluminescence.ROC curve was used to evaluate the diagnostic efficiency of sB 7-H1 alone or combined with other tumor markers in CRC .It also analyzed the correlation between serum sB 7-H1 level and some clinicopathological characteristics including tumor location , depth of invasion , lymph node metastasis status , distant metastasis status and tumor stages.Results (1)The differences of sB7-H1 level among CRC group, colorectal benign disease group and healthy control group ( compared with each other respectively ) were statistically significant ( P<0.001 ) . Levels of CEA, CA19-9, CA72-4 and CA-50 were significantly different between CRC group and benign colorectal disease group ( P<0.05 ) , while there was no significant difference between benign colorectal disease group and healthy control group (P>0.05).(2)The diagnosis ability of sB7-H1, CEA, CA19-9,CA72-4, and CA-50 were determined by the area under curve (AUC), which was 0.730, 0.772, 0.639, 0.663 and 0.635, respectively.The combination of sB7-H1 and CEA showed the best effect , and the AUC reached 0.831.(3)There was no significant correlation between the levels of sB 7-H1 and CEA, or CA19-9, CA72-4, CA-50.(4) Immunohistochemistry result showed that the positive rate of CRC group was 52％, while that of benign disease group was 6.3％.The difference of the positive rate between two groups was statistically significant (P<0.01).And there was a significant correlation between the positive rate of sB 7-H1 level in serum and tumor tissues of CRC patients ( P<0.05 ) .( 5 ) The serum sB7-H1 level in CRC patients was related to lymph node metastasis status (P<0.05).But there was no relationship between sB7-H1 level and other clinicopathological characteristics ( including tumor location , depth of invasion , distant metastasis status and tumor stage ) (P>0.05).Conclusion Serum sB7-H1 has a high clinical application value and could serve as a new tumor marker in CRC .

Objective To investigate the effectiveness and safety of thermal ablation of patients with 3－5 cm hepatocellular carcinoma ( HCC ) under the assistance of ultrasound fusion imaging . Methods From December 2010 to December 2017 ,76 HCC patients with 78 medium‐size ( 3 －5 cm ) lesions who underwent radiofrequency ablation ( RFA ) or microwave ablation ( M WA ) were included in the study . Ultrasound fusion imaging was used to assist the ablation procedures and assess the technical success immediately ,then guided supplementary ablation . Contrast‐enhanced CT/M R was performed one month after ablation to assess the technique efficacy rate . T he complication ,local tumor progression ( L T P ) rate were followed up . Results According to the ultrasound fusion imaging evaluation ,24 lesions ( 30 .8% , 24/78) received supplementary ablation immediately during the ablation procedure . Seventy‐one of 78 liver tumors were evaluated to achieve 5 mm ablative margin while the other 7 liver tumors were not achieved . T here were no ablation‐related deaths ,and the major complication rate was 6 .6% ( 5/76 ) .T he technique efficacy rate was 100% ( 78/78) according to the contrast‐enhanced CT/M R one‐month later . Patients were followed up from 6 to 79 months with a median time of 22 months . L T P occurred in 4 lesions and the LT P rate was 5 .1% ( 4/78) . Conclusions With the assistance of ultrasound fusion imaging ,thermal ablation of medium‐sized HCC is effective and safe w hich could achieve a higher technique efficacy rate and a lower L T P rate .

Objective:To summarize and analyze the risk of pregnancy recurrence of women with Duchenne muscular dystrophy (DMD) birth history in families with new DMD gene mutations, clarify the laws of DMD gene mutations and discuss the mode of genetic counseling in such families.Methods:Collected DMD families from January 2013 to December 2017 in Henan Provincial People′s Hospital. Firstly, the 79 exons of DMD gene were analyzed by multiplex ligation-dependent probe amplification (MLPA) in DMD patients and their mothers. The families that DMD patients with DMD gene mutations but no mutations in their mothers were selected for this study, and then MLPA combined with STR-gene linkage analysis were used to perform prenatal diagnosis for females in these DMD gene new mutation families.Results:A total of 64 families with new DMD gene mutations were included in this study. All mutations were DMD gene exon deletion mutations. A total of 65 fetuses were conducted prenatal diagnosis, included 26 SRY negative, 39 SRY positive; 63 fetuses′ DMD gene normal and 2 fetues′ DMD gene with exon deletion mutations. The results of postpartum follow-up and prenatal diagnosis were consistent.Conclusions:Exon mutations in newly mutated DMD families were mainly manifested as exon deletion, mainly presented in the 45-55 exon region. For families with new DMD mutations, even if there is no DMD gene mutation in women which had reproductive history of DMD, prenatal diagnosis for DMD during pregnancy was still recommended.

NK/T cell leukemia/lymphoma is a type of malignancy originating from T cells or natural killer cells with low incidence and poor clinical prognosis. There is still no effective treatment strategy. In recent years, targeted therapy has made great progress in the treatment of hematological malignancies, including monoclonal antibody and chimeric antigen receptor T cells (CAR-T), among which CD30, CD7, CD5, CD52, CCR4 and other target antigens are effective in the treatment of NK/T cell leukemia/lymphoma, but its widespread application still faces a great challenge. This article reviews the progress of immunotherapy for NK/T cell leukemia/lymphoma.

Objective To investigate the differences of gut microbiota in patients with schizophrenia between onset and remission. Methods Twelve patients with schizophrenia in the stage of onset were selected as the episode group. Thirteen gender-, age-and BMI-matched patients with schizophrenia during the remission period were selected as the control group. The fecal specimens of the two groups were collected for high-throughput sequencing of the 16S rRNA gene, and the gut microbiota differences between the two groups were analyzed. Results The chao index and ace index of gut microbiota was lower in the episode group than in the control group (t=2.385,P=0.026; t=3.068,P=0.005). The relative abundance of Bacteroides was higher and the relative abundance of Prevotella was lower in the episodes group than in the control group (Z=-2.013, P =0.044; Z=-3.427, P=0.001). The relative abundance of the Prevotella was negatively correlated with the PANSS score (r=-0.577, P=0.003). Conclusion Schizophrenia in the stage of acute episode have altered gut microbiota compared with those in the stage of remission and the gut microbiota is significantly corrected with mental symptoms.

Objective To conduct genetic diagnosis and prenatal diagnosis for a haemophilia B family with multi-nucleotides deletion mutation of F9 gene.Methods This is a genetic analysis.Whole exon mutation of the F9 gene was analyzed by PCR and Sanger sequencing for seven patients with the family of hemophilia B who consulted doctors in Henan Province People′s Hospital in April 2013.Suspected mutation was verified among non-hemophilia B members of the family and 100 healthy controls to rule out genetic polymorphism of the F9 gene.The above-mentioned detection results of hemophilia B gene , the pathogenic mutation of F9 gene in the family was clarified , and prenatal diagnosis was conducted for the female carriers in the family.It is recommended that the fetal gene detection should be conducted in amniotic fluid in the mid-term pregnancy of the female carriers of hemophilia , and then they can be informed of the non-hemophilia B fetus by the results of the gene detection .Results PCR and sequencing analysis has identified a deletion mutation of F9 gene c.185_188delGAGA[p.Glu62Asnfs?41]in seven hemophilia B patients.This mutation induced F9 gene frame shift mutation which led to early termination of F9 gene translation because there was a termination codon TAA at the 41th codon after the mutation site.The same mutation was not found among the non-hemophilia B members of the family and the 100 healthy controls. There were eight female carriers and nine female non-carriers in the family.Upon prenatal diagnosis , the Y chromosome sex-determining gene ( SRY ) in amniotic fluid was positive and no deletion mutation was observed in the F9 gene c.185_188.Conclusion The pathogenic mutation of F9 gene in the family was identified , which was helpful for prenatal diagnosis in female carriers .

OBJECTIVE： To study the effects of aspirin on the growth and autoghagy of human gastric cancer cells SGC-7901 and BGC-823. METHODS： SGC-7901 and BGC-823 cells were selected as research objects， with phosphate buffer （PBS） as negative control treated for 48 h， MTT assay was used to detect the effects of 1， 2， 4， 6， 8， 10 mmol/L aspirin， 5 mmol/L aspirin alone or combined with 2.5 μmol/L chloroquine， 2.5 μmol/L 3-methyladenine （3-MA） on survival rate of gastric cancer cells. Flow cytometry was used to detect the effects of 2 and 5 mmol/L aspirin， 5 mmol/L aspirin alone or combined with 2.5 μmol/L chloroquine and 2.5 μmol/L 3-MA on the apoptosis rate and cell cycle distribution of gastric cancer cells. Hoechst33258 staining was used to observe the effects of 5 mmol/L aspirin on morphology of gastric cancer cell nucleus； Transwell chamber test was adopted to detect the effects of 5 mmol/L aspirin on the migration of gastric cancer cell. Laser confocal scanning microscopy was used to observe the effects of 5 mmol/L aspirin on autophagy formation in gastric cancer cells. Western blot method was used to detect the effects of 2 and 5 mmol/L aspirin on the protein expression of autophagy markers LC3-Ⅱin gastric cancer cells. RESULTS： Compared with negative control group， aspirin could inhibit the survival rates of SGC-7901 and BGC-823 cells in dose-dependent manner， but had no significant effects on apoptosis rate of SGC-7901 and BGC-823 cells； SGC-7901 and BGC-823 cells were blocked in G1 phase. Compared with aspirin alone group， the survival rates of SGC-7901 and BGC-823 were increased significantly after treated with aspirin+chloroquine and aspirin+3-MA， while the distribution rate of SGC-7901 and BGC-823 cells at G1 phase were decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. Compared with negative control group， there were no obvious DNA fragmentation fragments， apoptotic bodies and fragments of dense bright blue， while the number of migration cells were decreased significantly in SGC-7901 and BGC-823 cells after treated with aspirin （P＜0.001）； the number of autophagosome was increased significantly and the protein expression of LC3-Ⅱ was enhanced significantly （P＜0.05）. CONCLUSIONS： Aspirin can significantly inhibit the growth of SGC-7901 and BGC-823 cells， and arrest cell cycle in G1 phase， the mechanism of which may be associated with the activation of autophagy.