BACKGROUND:The mechanism of differentiation and proliferation of bone marrow-derived mesenchymal stem cells remains unclear. In addition, issues such as how signal pathways such as Ca2+and bone marrow-derived mesenchymal stem cellproliferation and differentiation signals form complex signal network remain poorly understood.
OBJECTIVE:To investigate the effect of Ca2+in the induced differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes.
METHODS:Bone marrow-derived mesenchymal stem cells were isolated from rat bone marrow using whone bone marrow adherence method, purified, amplified, and induced with hepatocyte growth factor. [Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells and control bone marrow-derived mesenchymal stem cells were detected with flow cytometry. Bone marrow-derived mesenchymal stem cells induced with hepatocyte growth factor were mixed with nimodipine of different concentration, and cells were divided into three groups:hepatocyte growth factor+nimodipine 10 mg/L, 50 or 100 mg/L groups. cellgrowth was observed with inverted phase contrast microscope and alpha 1-antitrypsin expression of the cells was confirmed by immunocytochemistry. The calcineurin M and the activation of extracellular signal regulated kinase pathway was detected by reverse transcription-PCR and western blotting, respectively.
RESULTS AND CONCLUSION:[Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells was higher than in the control group (P<0.05). After addition of a larger dose of nimodipine, no differentiation of cells was obeserved and growth of bone marrow-derived mesenchymal stem cells was getting worse. There were few alpha 1-antitrypsin positive cells in the nimodipine groups. Calcineurin Mexpression was significantly increased in directional differentiated bone marrow-derived mesenchymal stem cells and smal dose of nimodipine than the controls (P<0.05). However, no significant difference was found among middle, high dose nimodipine and control groups (P>0.05). These findings indicate that Ca2+could participate in the differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes incuded with cytokines, and also maintain the survival and proliferation of bone marrow-derived mesenchymal stem cells.

Objective To determine the difference in blood microcirculation recovery between normal frostbite and high-altitude frostbite during the wound healing.Methods Twenty four male rats were randomly divided into control group (n=8), normal frostbite group (n=8), and high-altitude group (n=8). The normal frostbite group rats were frozen to produce mid-degree frostbite models by controlling the freezing time with liquid nitrogen penetration equipment. The high-altitude frostbite group rats were acclimated to a hypoxic and low-pressure environment for 1 week, and then the high-altitude frostbite models were constructed by the same way with liquid nitrogen penetration apparatus. On days 3, 7, 11, 15, 19, and 23 after modeling, the recovery situation of blood circulation of each group was observed with contrast ultrasonography by injecting SonoVue micro-bubble into rats' tail. Finally, the micro-bubble concentration (MC) was calculated to confirm the blood circulation recovery with software Image Pro. ResultsAt different time points, the wound area of the high-altitude frostbite group was bigger than that of the normal frostbite group, and the MC of control group was always about (27±0.2)×109/ml. On day 3, 7, 11, 15, 19, and 23, the MC was significantly lower in the high-altitude frostbite group than in the control group and normal frostbite group (P<0.05). The MC of normal frostbite group was significantly lower than that of the control group on day 3, 7, 11, 15 and 19 (P<0.05). In addition, no obvious difference in MC was found between normal group and control group on the 23th day (P<0.05).Conclusion The blood microcirculation recovery after high-altitude frostbite is significantly slower than the normal frostbite.

Aim To investigate the effect of sphingo-sine kinase 1 (SphK1 )on unilateral ureteral obstruc-tion(UUO)-induced tubulointerstitial fibrosis and ex-plore the possible mechanism.Methods The CD-1 mice were randomly divided into four groups:sham-op-eration group(Sham),PF-543 treatment control group (Sham +PF-543),model group(UUO)and PF-543 treatment group(UUO +PF-543).On 1 ,3,7 and 1 4 d after operation,eight mice were selected randomly from each group and sacrificed.The protein expressions of SphK1 ,mature TGF-β1 ,FN,ColⅠ,LC3,Beclin1 ,Atg5 and Atg1 2 were observed by Western blot.The histo-logical changes were examined by Masson′s trichrome stain.Immunhistochemistry was performed to measure the levels of expression of SphK1 ,FN and Col Ⅰ. Transmission electron microscope was used to observe the autophagic body.Results SphK1 expression and autophagy were both upregulated in a mouse model of kidney fibrosis induced by UUO. Meanwhile, in-creased mature TGF-β1 and deposition of extracellular matrix(ECM)were observed in tubulointerstitial areas compared with sham-operated mice.After intraperito-neal injection with the SphK1 specific inhibitor PF-543 in UUO mice,enhanced expression of SphK1 and acti-vated autophagy were significantly abrogated.Howev-er,aggravation of renal fibrosis was detected when SphK1 inhibitor PF-543 was applied to suppress SphK1 expression in UUO mice.Conclusion SphK1 activa-tion is renoprotective through the induction of autoph-agy in the pathogenesis of kidney fibrosis.

To determine the distribution and drug resistance tendency of Gram-negative bacillus so as to provide a reasonable selection for antibiotic in clinical treatment.The clinical isolates bacillus were mainly Gram-Negative bacillus during three years.The detection out ratio for Gram-Negative bacillus had decreased gradually but the ratio for non-fermental gram-negative bacillus had increased.The combinations of antibiotic and enzyme inhibitor has more antibacterial activity to non-fermental gram-negative bacillus than antibiotic.The detection out ratio for ESBLs had gradually increased.

Objective To investigate the expressions of FAT10 and p53 mutant in gastric cancer tissues and their relations. Methods Immunohistochemistry and RT-PCR were used to detect the expressions of FAT10 and p53 in gastric cancer tissues (n=62), para-cancerous tissues (2-5 cm apart from cancer, n=62), and normal gastric tissues (7>5 cm apart from cancer, n=62). The association of FAT10 with p53 and clinical outcomes were analyzed by Spearman and Pearson correlation. Results The immunohistochemistry examination showed that expressions of FAT10 [51.61%(32/62)] and p53 [45.16% (28/62)] were significantly higher in cancerous tissues than in para-cancerous tissues [12.90%(8/62) and 14.51% (9/62), χ2=21.26 and 20.69, P<0.01] and normal tissues [6.45% (4/62) and 9.68% (6/62), χ2=13.91 and 19.61, P<0.01]. Overexpressions of FAT10 protein and mRNA in cancerous tissues were closely related to lymph node metastasis and TNM staging (both P value<0.05). There was a positive correlation between FAT10 and p53 in protein and mRNA expressions (protein r=0. 865, P<0.05; mRNA r=0.761, P< 0.01). Those with positive expression of FAT10 had lower survival rate compared to those with negative expression (P<0.05). Conclusions The positive relation between over-expression of FAT10 and p53 implicates that both are involved in the gastric carcinogenesis, and FAT10 is a novel gastric cancer marker with prognostic significance.

Objective To evaluate the clinical value of endoscopically nasojejunal feeding tube placement (ENFTP) for nutritional support in patients with severe acute pancreatitis (SAP). Methods Those SAP patients who treated with ENFTP (n= 47) or with total parenteral nutrition (TPN) (n=50) were retrospectively analyzed for laboratory parameters before and 1,2 and 4 weeks after nutrition support. Outcomes in the two groups were compared with respect to complications,mortality, duration of feeding, feeding costs,mechanical ventilation time and length of ICU or hospital stay. Results Four weeks after nutrition sypport, the hemoglobin and albumin were increased in ENFTP group as compared to TPN groups (P < 0. 05), while the blood sugar was decreased significant in ENFTP group than in TPN group (P<0.05). The incidence of peripancreatic or biliary infection and catheter-related infection were lower in ENFTP group than in TPN group. Duration of feeding and hospital stay were shorter, and nutrition cost was lower in ENFTP group (P<0.05). In addition, the APACHE Ⅱ score was significantly improved in ENFTP group(P<0.05). Conclusion ENFTP seems to be safe and less expensive in treatment of patients with acute pancreatitis.

OBJECTIVETo investigate the hemodynamics evidence of the descending branch of lateral circumflex femoral artery in a reversed way. To explore the clinical result of using the reversed descending branch of the lateral circumflex femoral artery as the receipt artery for free flaps for reconstruction of the leg soft-tissue defect.

METHODSFrom October 2005 to February 2012, 38 patients with severe leg soft-tissue defects were treated. The proximal antegrade and retrograde mean artery pressure of the descending branch of the lateral circumflex femoral artery in 16 of 38 patients were recorded during operation. All wounds had osteomyelitis, bone and tendon exposure requiring coverage reconstruction. And there was no recipient artery in the injured lower leg for free flaps in all 38 patients. Reversed descending branches of lateral femoral circumflex arteries were used as recipient arteries for free flaps (free latissimus dorsi flap, free thoracoumbilical flap, and free anterolateral thigh flap) in all patients. The flap donor site was closed directly or with the skin graft.

RESULTSThe proximal antegrade mean artery pressure of the descending branch of lateral circumflex femoral artery was(81.6 +/- 12.4) mmHg. The proximal retrograde pressure was(48.2 +/- 10.7) mmHg. The proximal retrograde mean artery pressure was 59.07 percent of the proximal antegrade pressure. The donor skin graft survived and wound healed primarily. After operation, 2 flaps had distal partial necrosis and healing was achieved after dressing change. All the other flaps survived completely without vascular problems. All the patients were followed up for 11 months to 2.5 years (mean, 1.6 years). The flap appearance was satisfactory. The texture and color of flaps in all cases were good.

CONCLUSIONSThe reverse descending branch of lateral circumflex femoral artery is a reliable recipient artery for the free flaps. It is an easy and simple technique that can be used for reconstruction of the defects in the lower leg, with the reversed descending branch of lateral circumflex femoral artery as recipient artery.

Adolescent ; Adult ; Blood Pressure ; Female ; Femoral Artery ; physiopathology ; surgery ; Free Tissue Flaps ; blood supply ; Hemodynamics ; Humans ; Lower Extremity ; injuries ; Male ; Middle Aged ; Soft Tissue Injuries ; surgery ; Young Adult

Objective Implement etimicin polymethylmethacrylat (PMMA) bone cement implantation treatment for the postoperative patients suitable with intramedullary infection after open fracture surgery,and observe the efficacy,provide new ideas for clinical treatment.Methods Selected 60 patients in Second People's Hospital of Kunshan from Jan.2009 to Dec.2016 whose infection was confined to the fracture caused by trauma after the internal fixation.Thirty cases of etimicin bone cement implantation were treated group,30 cases of CEMEX GEN antibiotic bone cement (containing gentamicin) implantation were control group.The internal fixation was taken out of 9 cases of autogenous PMMA bone graft with bone cement cover in the treatment group,debrided and taken first phase of bone grafting internal fixation combined with etimicin PMMA bone cement,21 patients with direct bone cement filling underwent surgery to remove the internal fixation devices,debridement cavity filling etimicin bone cement.Six months later when the infection was controlled,the second phase of autogenous bone graft fixation was done.Ten cases of autogenous bone graft with bone cement cover in the control group,debrided and taken first phaes of bone grafting internal fixation combined with CEMEX GEN antibiotic bone cement (containing gentamicin).Twenty patients with direct bone cement filling underwent surgery to remove the internal fixation devices,debridement cavity filling CEMEX GEN antibiotic bone cement (containing gcntamicin).Six months later when the infection was controlled,the second phase of autogenous bone graft fixation was done.SPSS 19.0 software was used for statistical analysis.The t test was used for measurement data,and the counting data were checked by chi-square.Results The pathogens were mainly staphylococcus aureus and coagulase-negative staphylococci.Thirty-seven strains of pathogenic bacteria were detected with MIC ≤ 2 mg/L in the treatment group,which was in the safe and effective drug concentration range of etimicin(MIC ≤ 12 mg/L).Thirty strains of pathogenic bacteria were detected with MIC ≤ 8 mg/L in the control group,which was in the safe and effective drug concentration range of gentamicin(MIC ≤ 12 mg/L).The average follow-up period of the treatment group was 11.2 months (range 8 to 24 months),29 cases were controlled.The average follow-up period of the control group was 15.6 months (range 6-35 months),24 cases were controlled.The difference of infection control rate between treatment group and control group was statistically significant (x2 =4.043,P =0.044).Conclusion The complete debridement,a lot of washing,and etimicin bone cement implantation can effectively control the infection after the internal fixation of fracture and reduce the recurrence,which can be used in clinical practice.

In order to establish the stable andreliable ISSR-PCR System of Lysimachia christinae, L16 (4(5)) orthogonal design, which based on 7 levels of single factor experiment, were used in this study. The variance analysis was carried out by SPSS 19.0, and 5 main factors affecting the reaction system were optimized in 4 levels. The best annealing temperature was selected by the optimized reaction system. And the stability and reliability of this system was tested by 23 samples from different origins. The results showed that the five factors (DNA template, primer, dNTP, Mg2+ and Taq enzyme) were the most impacts on the amplified results of ISSR-PCR of L. christinae. The order of the influence was: primer > Taq enzyme > DNA template > Mg2+ > dNTP. The optimal system, which was determined by multiple comparison on different levels of each factor, was total volume of 25 microL, including DNA template 60 ng, primer 0.3 micromol x L(-1), dNTP 0.2 mmol x L(-1), Mg2+ 1.8 mmol x L(-1), Taq enzyme 1.25 U. The optimal system was stable and reliable tested by 23 samples from different origins. This study lays the foundation for genetic diversity analysis, fine varieties selection and molecular identification of L. christinae, and provides reference for optimization on ISSR-PCR system of other speciesin future.
DNA Primers ; genetics ; DNA, Plant ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Microsatellite Repeats ; Polymerase Chain Reaction ; methods ; Primulaceae ; classification ; genetics ; Quality Control

OBJECTIVETo explore the association between chromosomal disequilibrium and chemoresistance/chemosensitivity in non-small cell lung cancer (NSCLC) using comparative genomic hybridization (CGH).

METHODSGenomic DNA samples were prepared from the tumor tissues in paraffin-embedded sections derived from 88 patients with advanced NSCLC (18 with chemosensitivity and 16 with chemoresistance). The DNAs were first amplified by a degenerate oligonucleotide prime-polymerase chain reaction protocol and then labeled with fluorescence as probes for CGH analyses. The correlations of the resulting chromosomal imbalances with the chemo-sensitivity and other pathological features of the patients were analyzed.

RESULTSA total of 640 abnormal chromosome regions including 96.12% gains and 3.88% losses were detected in 88 specimens. The results indicated that the most frequently gained chromosome regions were 19p13.1-13.3 (39/88, 44.12%), followed by 9q12-q22 (26/88, 29.41%), 22q12-q13 (26/88, 29.41%), and Xq (29/88, 32.35%). The total number of abnormal regions related with chemo-sensitivity was 188( 182 gains and 6 losses), while the number of the abnormal regions linked to the chemoresistance was 452 (431 gains and 21 losses) (P=0.005). Gains of 14p12-p13 and 19p were significantly correlated with the chemosensitivity of the NSCLC (P=0.006). Gains of 1q12-q22, 10q25-q26, 5p15.1-p15.3, 19q13.2-13.4, 20p11.2-p12, 21q22, and Xp 21-p22.1 were also significantly correlated with the chemoresistance (P]0.005, 0.029, 0.039, 0.029, 0.039, 0.016, and 0.006, respectively). No correlation between the chromosome abnormalities and other clinical features was observed.

CONCLUSIONSThe specific gains and losses of chromosome region is correlated with platinum-based first-line chemotherapy in NSCLC patients,as confirmed by CGH detection. This finding is useful for further identifying the chemosensitivity-related functional genes, predicting clinical effectiveness, and achieve individualized treatment in the future.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; Chromosome Aberrations ; Comparative Genomic Hybridization ; Drug Resistance, Neoplasm ; genetics ; Female ; Humans ; Karyotyping ; Lung Neoplasms ; drug therapy ; genetics ; Male ; Middle Aged ; Treatment Outcome