Objective To explore the predictive value of microvessel density(MVD)and blood vessel invasion(BVI)in hepatic metastasis from early-stage rectal cancer.Methods MVD and BVI in the tumor tissue from 380 patients with stage I and II rectal cancer was determined by immunohistochemical S-P method with anti-CDIOS antibody and anti-CD34 antibody,respectively.Multinomial logistic regression was performed to analyze the predictive value of MVD and BVI in hepatic metastasis from early-stage rectal cancer.Results CD105 was expressed in newborn blood vessels,not in normal blood veseels.in the rectal cancer tissue.MVD was correlated with histological type and infiltration depth(P＜0.05).Besides histological type and infiltration depth,BVI was also correlated with histological grade.Multivariate analysis revealed that histological type,tumor infiltration depth,BVI,adjuvant therapy,and MDV were independent predictors of hepatic metastasis from rectal cancer.The risk of hepatic metastasis in patients with postive expression of either MVD or BVI or both were significant higher than that in patients with low expression of MVD and those without BVI expression[hazard ratio(95%CI),4.210(2.182-11.214)].Conclusion BVI and MVD are independent predictors of hepatic metastasis from stage I and II rectal cancer.Combined detection of MVD and BVI may help to predict the clinical outcome of patients with early-stage rectal cancer.

BACKGROUND:Reducing the ischemia-reperfusion injury in the system of simulated in vivo physiological environment for amputated limbs, can extend the preservation time and improve the replantation success rate of amputated limb. OBJECTIVE:To investigate the effect of Mailuoning on the ischemia-reperfusion injury in the system of simulated in vivo physiological environment for amputated limbs. METHODS:The left hind legs harvested from 18 healthy adult male Bama mini-pigs were randomly divided into three groups:cold storage group, blood perfusion group, Mailuoning combined blood perfusion group, with six pigs in each group. After the left hind legs of each pig in al groups were amputated and stored at room temperature for 3 hours, the amputated legs were placed at 4 ℃, perfused with blood, or perfused with Mailuoning combined blood. After 6 hours of perfusion, the morphology of amputated limbs was observed under transmission electron microscope. The mRNA levels of apoptosis proteins Caspase3 and inflammatory factor interleukin-1βwere detected by real time fluorescent quantitative RT-PCR.RESULTS AND CONCLUSION:Transmission electron microscopy results showed that, the muscle fibers in blood perfusion group and Mailuoning combined blood perfusion group arranged more orderly and were more complete than cold storage group, the swel ing of mitochondria was lighter. In addition, the condition in Mailuoning combined blood perfusion group was better than in blood perfusion group. RT-PCR results showed that the expression of Caspase3 and interleukin-1βwas increased in al groups;at the same time, the expression level in blood perfusion group and Mailuoning combined blood perfusion group was significantly lower than those in cold storage group. Mailuoning combined blood perfusion group had s lower expression than blood perfusion group. In the system of simulated in vivo physiological environment for amputated limbs, Mailuoning can significantly reduce the ischemia/reperfusion injury of skeletal muscle cells and prolong the preservation time of amputated limbs.

Objective: To investigate the effectiveness of local injection of autologous platelet-rich plasma (PRP) in treatment of diabetic foot ulcer. Methods: Between October 2017 and October 2018, 90 diabetic foot ulcer patients who met the selection criteria were randomly divided into 3 groups: PRP injection group (group A, PRP was injected and hydrogel dressing covered the wounds), PRP covered group (group B, PRP gel and hydrogel dressing covered the wounds), and the control group (group C, hydrogel dressing covered the wounds), 30 cases in each group. There was no significant difference in gender, age, injured side, disease duration, preoperative glycosylated hemoglobin, wound size, and Wagner grading between groups (P>0.05). The frequency of treatments and hospitalization day in all groups and the total amount of PRP application in groups A and B were recorded. The wound healing condition was recorded during the treatment, and the wound healing rate was calculated at 3 months after the first debridement. Results: The frequency of treatments in groups A, B, and C were (10.2±0.8), (11.4±0.6), (12.5±0.5) times, respectively. The total amount of PRP application of groups A and B were (306±24) and (342±18) mL, respectively. There was no significant difference in the frequency of treatments and the total amount of PRP application between groups (P>0.05). The hospitalization days of groups A, B, and C were (40.5±1.8), (62.1±2.3), and (88.6±1.4) days, respectively, showing significant differences between groups (P<0.05). In the course of treatment, the necrosis and exudation of the wounds gradually reduced, the areas of wounds gradually reduced; and the above conditions of group A were significantly better than groups B and C, and group B was better than group C. At 3 months after the first debridement, the wound healing rates of groups A, B, and C were 93.2%±0.8%, 52.1%±1.1%, and 21.3%±1.3%, respectively, with significant differences between groups (P<0.05). Conclusion: PRP can effectively promote the repair of diabetic foot ulcer. The effectiveness of local injection of PRP is superior to the local coverage.

Carbocisteine is a commonly used expectorant drug.Currently,the reported detection methods for carbocisteine mainly include the electrochemical method,potassium bromated titration,kineticmethod,spectrometry,chromatography,HPLC-FLD precolumn derivation determination,HPLC-MS,etc.The advantages and disadvantages of each method were compared and analyzed in this paper,and it is found that HPLC was more suitable for the quality analysis of carbocisteine,which could serve as a reference for upgrading standard of carbocisteine content determination.

Objective To investigate the role of cell adhesion molecule L1 like (CALL) in the genesis and development of esophageal squamous cell carcinoma (ESCC).Methods From July 2007 to December 2010,a total of 100 patients with ESCC who received radical resection of esophageal cancer were enrolled.The ESCC tissues and corresponding tumor-adjacent normal tissues were obtained.The expression of CALl was determined by tissue microarray technology and immunohistochemical staining.The CALL over-expressed esophageal cancer cell line was established.The effects of CALL on cell migration and invasion were detected by wound-healing assay and Transwell assay,respectively.The effects of CALL on actin microfilament was analyzed by filamentous actin (F-actin) staining.Chi square test,Fisher's exact test,multivariate analysis and t test were performed for statistical analysis.Results The positive expression rate of CALL in ESCC tissues was 56 ％ (56/100),which was lower than that of tumor-adjacent normal tissues (95％,95/100),and the difference was statistically significant (x2=41.114,P＜0.01).There were statistically significant differences in CALL expression at protein level among patients with ESCC of different differentiation degree,different pathological T stage,lymph node metastasis and different TNM stage (x2=13.702,5.317,21.453,Fisher's exact test;all P＜ 0.05).The five year disease related survival rate of ESCC patients with down-regulated expression of CALL was 0(0/49),which was lower than those with normal CALL expression (25.5％,13/51),and the difference was statistically significant (x2 =43.338,P＜0.01).The median survival time of CALL expression down-regulated group was 17 months,and that of normal expressed group was 38 months.CALL expression was an independent risk factor of disease special survival rate (hazard ratio (HR) 0.353,95％ confidence interval (CI) 0.188 to 0.666,P=0.001).The results of wound-healing assay showed that the migration ability of CALL overexpressed CALL-k30 cells was lower than that of Vec-k30 cells in control group on 24 hours after wound.The results of Transwell invasion test showed the number of migrating cells penetrating CALL k30 cells attached to the inferior surface of the membrane was 44.000±13.748,which was less than that of the Vec k30 cells (154.333±25.007),and the difference was statistically significant (t=5.136,P=0.036).The results of F-actin staining demonstrated that actin filaments of CALL-k30 cells was 234.667 ± 65.118,which was lower than that of Vec-k30 cells (597.000± 119.929),and the difference was statistically significant (t=4.707,P=0.042).Conclusions CALL lowers the migration and invasion abilities of esophageal cancer cells by inhibiting F-actin microfilaments.Its abnormal expression may play an important role in the genesis,development and prognosis of ESCC.

Objective: To investigate the expression of ALC1 protein during esophageal squamous cell carcinoma (ESCC) development and progression, so as to explore its association with clinicopathological characteristics and overall survival of ESCC patients, and the effect of ALC1 overexpression on malignant biological behavior of esophageal squamous cells. Methods: Immunohistochemistry (IHC) was used to detect ALC1 protein expression in 245 primary ESCC tissues and their paired normal esophageal mucous membranes, and to determine its correlation to gender, age, tumor cell differentiation, invasion, TNM stage, lymph nodes metastasis, and overall surviv-al rate of ESCC patients. MTT assay, colony formation assay, transwell invasion, and wound healing assay were used to observe the ef-fect of ALC1 on ESCC cell proliferation, invasion, and migration. Results: The expression ratio of ALC1 in esophageal squamous cell car-cinoma was higher compared with that in their paired normal esophageal mucous membranes (41.6% vs . 21.2% , P<0.05). Upregula-tion of ALC1 was associated with ESCC invasion, TNM stage, and lymph node metastasis (P<0.05). The overall survival of ESCC patients with ALC1 overexpression was significantly lower than that in patients with downregulated ALC1 expression (P=0.002). Therefore, ALC1 may promote the proliferation, invasion, and migration of ESCC cells. Conclusions: ALC1 upregulation may play an important role in the progression and development of ESCC. Upregulation of ALC1 leads to poorer disease prognosis, and could promote the prolifera-tion, invasion, and migration of the KYSE30 ESCC cells. Therefore, ALC1 may have potential prognostic value for ESCC patients.

Objective:To explore the role and molecular mechanism of hepatocyte nuclear factor 4γ (HNF4γ) in proliferation and stemness of gastric cancer.Methods:A total of 102 cases of paraffin-embedded gastric cancer tissues and matched adjacent gastric tissues and 42 cases of fresh-frozen tissues derived from gastric patients who received radical gastrectomy were collected from the First Affiliated Hospital of Zhengzhou University between 2012 to 2015. The expression of HNF4γ was tested by immunohistochemical staining, quantitative real-time polymerase chain reaction (qRT-PCR). HNF4γ overexpressed (AGS-HNF4γ) and shRNA silenced (HGC27-shHNF4γ) gastric cell lines were established. The effects of HNF4γ on cell proliferation and stemness were verified by XTT, clone formation and sphere formation assay. The expression of CD44 was detected by western blot.Results:The mRNA expression level of HNF4γ in fresh-frozen gastric cancer tissue was (12.43±2.702), which was significantly higher than (3.639±1.109) in normal tissue ( P<0.001). The high protein expression rate of HNF4γ in paraffin-embedded gastric cancer tissues was 41.2% (42/102), which was significantly higher than 8.8% (9/102) in normal gastric mucosa tissue ( P< 0.001). The protein expression of HNF4γ was closely related to the tumor differentiation, infiltration depth, lymph node metastasis and tumor stage ( P<0.05). The median survival interval of patients with HNF4γ high expression was 25 months, the 3-year survival rate was 4.8% (2/42), significantly lower than 38 months and 51.7% (31/60) of patients with normal HNF4γ expression ( P<0.001). The proliferation and CD44 protein expression of AGS-HNF4γ cells were significantly higher than those of the AGS-Vector cells. The number of clone formation, sphere formation rate of AGS-HNF4γ cells were 243.5±24.5 and (83.5±3.9)%, significantly higher than 81.0±16.0 and (21.8±5.6)% of AGS-Vector cells ( P=0.030 and P=0.010, respectively). The proliferation and CD44 protein expression of HGC27-shHNF4 cells were significantly lower than those of the HGC27-vector cells. The number of clone formation, sphere formation rate of HGC27-shHNF4 cells were 26.0±1.0 and (20.8±8.4)%, significantly higher than 83.5±4.5 and (72.5±4.8)% of HGC27-vector cells ( P=0.006 and P=0.030, respectively). Conclusions:HNF4γ is upregulated in the gastric cancer tissues and related with the poor prognosis of patients with gastric cancer. Overexpression of HNF4γ promotes the proliferation and remains the stemness of gastric cancer cells by upregulating the expression of CD44.

Objective:To explore the role and molecular mechanism of hepatocyte nuclear factor 4γ (HNF4γ) in proliferation and stemness of gastric cancer.Methods:A total of 102 cases of paraffin-embedded gastric cancer tissues and matched adjacent gastric tissues and 42 cases of fresh-frozen tissues derived from gastric patients who received radical gastrectomy were collected from the First Affiliated Hospital of Zhengzhou University between 2012 to 2015. The expression of HNF4γ was tested by immunohistochemical staining, quantitative real-time polymerase chain reaction (qRT-PCR). HNF4γ overexpressed (AGS-HNF4γ) and shRNA silenced (HGC27-shHNF4γ) gastric cell lines were established. The effects of HNF4γ on cell proliferation and stemness were verified by XTT, clone formation and sphere formation assay. The expression of CD44 was detected by western blot.Results:The mRNA expression level of HNF4γ in fresh-frozen gastric cancer tissue was (12.43±2.702), which was significantly higher than (3.639±1.109) in normal tissue ( P<0.001). The high protein expression rate of HNF4γ in paraffin-embedded gastric cancer tissues was 41.2% (42/102), which was significantly higher than 8.8% (9/102) in normal gastric mucosa tissue ( P< 0.001). The protein expression of HNF4γ was closely related to the tumor differentiation, infiltration depth, lymph node metastasis and tumor stage ( P<0.05). The median survival interval of patients with HNF4γ high expression was 25 months, the 3-year survival rate was 4.8% (2/42), significantly lower than 38 months and 51.7% (31/60) of patients with normal HNF4γ expression ( P<0.001). The proliferation and CD44 protein expression of AGS-HNF4γ cells were significantly higher than those of the AGS-Vector cells. The number of clone formation, sphere formation rate of AGS-HNF4γ cells were 243.5±24.5 and (83.5±3.9)%, significantly higher than 81.0±16.0 and (21.8±5.6)% of AGS-Vector cells ( P=0.030 and P=0.010, respectively). The proliferation and CD44 protein expression of HGC27-shHNF4 cells were significantly lower than those of the HGC27-vector cells. The number of clone formation, sphere formation rate of HGC27-shHNF4 cells were 26.0±1.0 and (20.8±8.4)%, significantly higher than 83.5±4.5 and (72.5±4.8)% of HGC27-vector cells ( P=0.006 and P=0.030, respectively). Conclusions:HNF4γ is upregulated in the gastric cancer tissues and related with the poor prognosis of patients with gastric cancer. Overexpression of HNF4γ promotes the proliferation and remains the stemness of gastric cancer cells by upregulating the expression of CD44.

Objective To investigate the clinical application and nursing of maggot debridement therapy in the diabetic foot , pressure ulcer and other refractory wounds .Methods Eleven patients with refractory wounds were selected .We used maggot debridement therapy for wound debridement , and implemented considerate clinical care and psychological care for their respective basic diseases .Results The mean healing time of the refractory wounds was (25.0 ±3.8) days.The patients received scientific and considerate nursing and accepted the biological debridement therapy psychologically .Conclusions For the treatment of chronic refractory wounds , maggot debridement therapy is an effective , economical and safe method .

Objective To evaluate the effects of the multiple factors especially image geometric accuracy of the imaging system on the segmentations of target areas and organs-at-risk.Methods The study used phantoms to test the imaging performance of the 1.5T magnetic resonance(MR)linear accelerator system,including the assessments of MR image geometric distortion and the segmentation errors caused by factors such as image geometric distortion.Model 604-GS large field MR image distortion phantom was used to explore the geometric distortion of the MR images for MR-guided radiotherapy;and CIRS Model 008z upper abdominal phantom was used to analyze the segmentation errors of target areas and organs-at-risk.Results The average geometric distortion and maximum distortion of 3D T1WI-FFE images vs 3D T2WI-TSE images were 0.54 mm vs 0.53 mm and 1.96 mm vs 1.68 mm,respectively;and the control points of the large distortions were distributed at the edges of the phantom,which was consistent with the MR imaging characteristics previously reported.Compared with CT-based segmentation contour,the MDA was 1.17 mm and DSC was 0.91 for 3D T1WI-FFE,while MDA was 0.86 mm and DSC was 0.94 for 3D T2WI-TSE.Conclusion The study quantitatively assesses the geometric accuracy of the imaging system for MR-guided radiotherapy.The phantom-based contour analysis reveals that with CT image as gold standard,the segmentation error in MRI images meets the clinical requirements,and that 3D T2WI-TSE image is advantageous over 3D T1WI-FFE image in segmentation accuracy.