Objective To observe effects of rhubarb extract on Ca dissolved quantity in demineralized enmnel. Methods Demineralization was performed after treating tooth facing with rhubarb extract of different concentrations and deionized water. The concentration of Ca was detected by ASCA biochemical analyzer after demineralization. Results Dissolved quantity of demineralized Ca in the rhubarb extract group (2 mg/ml, 4mg/ml) was significantly lower than that of the deionized water group (P<0.01) and the rhubarb extract group (1 mg/ml) (P<0.05), but higher than that of 2% sodium fluoride group (P>0.05). Dissolved quantity of demineralized Ca in the rhubarb extract (1 mg/ml) group was significantly higher thanthat of 2% sodium fluoride group (P<0.01), but lower than that of the deionized water group(P>0.05).Conclusion The rhubarb exwact (2mg/ml, 4mg/ml) can inhibit Ca dissolution in demineralized enamel.The inidal effective concentration of rhubarb extract is 2 mg/ml.

Objective To investigate the effect of salvia miltiorrhiza (SM) on the expression of Bcl 2?Bax and hepatocyte apoptosis in hepatic preservation and reperfusion in a murine model.Methods The model of the preservation reperfusion of isolated rat liver was established. Bcl 2?Bax expression of the liver tissue and hepatocyte apoptosis were studied respectively by flow cytometry and in situ terminal deoxynucleotidyl transferase mediated dUTP FITC nick end labeling (TUNEL) technique. Results In SM treated group levels of Bcl 2 expression was significantly increased ( P

Objective To investigate migration of dendritic cells(DC)in orthotopic rat liver transplantation models.Methods Allogeneic models(Wistar→SD,experimental group)and syngenic models(Wistar→Wistar,control group)of rat liver transplantation were established.Graft livers and host celiac lymph nodes(n=4)of each group were sampled respectively at day 3,5,and 7 after the transplantation.The acute rejection was graded according to liver histopathological changes.The dy- namic state of DC number within graft and lymph nodes was detected by means of immunohistochemi- cal staining and image analysis.T-cell active proliferative response in lymph nodes was also studied. Results The histological examination revealed that mild to severe rejection occurred on the post-opera- tive days 5 and 7.At day 3 after transplantation,the number of S-100~+ DC in allograft was signifi- cantly increased and reached the peak at day 5,then decreased gradually at day 7.A significant num- ber of S-100~+ cells was detected in the allogeneic host lymph nodes from day 3 after transplantation, and displayed a continuous increasing trend for next several days.Active proliferation of T cells in the lymph nodes was triggered as early as day 3 after transplantation.Conclusion Allogeneic liver trans- plantation induces the accelerated migration of DC within the allograft and host lymphoid nodes.DC delivers a strong and sustained stimulation among T cells in the lymphoid nodes where effector cells are predominantly sensitized and rejected the graft eventually.

Objective To observe the survival rate of free flaps and incidence of vascular crisis following the perforator flap transfer in end-to-side microvascular anastomosis compared with the traditional method and investigate the scientificity and practicability of this scheme.Methods From February, 2012 to October, 2013, 152 cases of free flaps (including superficial illiac circumflex perforator flap, lateral arm flap, peroneal perforator flap, and anterolateral thigh flap) were performed.The end-to-side anastomosis were performed in 72 cases while end-to-end anastomosis in 80 cases;All patients were followed up for 6-24 months.The occurrence of vascular crisis in all cases were observed and recorded.The results of the two groups were conducted statistical analyzed.Results The average anastomosis time for end-to-side anastomosis (21.3 ± 3.8 min) was significantly higher than the end-to-end anastomosis (14.4 ± 3.2 min) (P ＜ 0.05).The incidence of anastomotic vasospasm after end-to-side anastomosis (5.6％) was significantly lower than traditional method (16.3％) (P ＜ 0.05).There was no significant difference in the survival rate of the free flaps between the end-to-side anastomosis group (97.2％) and the traditional group postoperatively (96.3％) (P ＞ 0.05).Conclusion Although anastomosis times were increased in the end-to-side group, this technique showed lower spasm rate and similar flap survival rate.Therefore, this technique is a reliable and technically feasible method and should be considered first as a choice when facing vessel discrepancy and for preserving the recipient artery and vein system.

Objective:To investigate the diagnostic value of MRI and CT for the liver space-occupying lesions.Methods:The clinical data of 70 cases of patients with liver space-occupying lesions in our hospital from June 2012 to May 2016 were divided into two groups and retrospectively analyzed.35 cases underwent contrast enhanced ct scans (CT group),and others underwent dynamic contrast-enhanced MR imaging(MRI group).The pathological diagnosis,number of lesions and lesions diameter were ompared between two groups.Results:No significant difference was found in the pathological diagnosis,number of lesions(71 vs 70) and lesions diameter(2.25 ± 2.01 cm vs 2.19± 1.98 cm) between two groups(P＞0.05).As the gold standard by pathological diagnosis results,correct diagnostic rate of MRIgroup were 85.71％,which was 77.14％ CT group and lower than that of the MRI group,but no significant difference was found between two groups (P＞0.05).The incidence of adverse reactions in CT group was significantly higher than that of the MRI group (P＞0.05).Conelusion:Both CT and MRI enhancement scanning have higher diagnostic value for the liver space-occupying lesions,but MRI enhancement scanning has higher safety and tolerability.

BACKGROUND: Bone morphogenetic protein 2 is a most widely studied and osteogenic-inducing bone morphogenetic protein. However, simple bone morphogenetic protein can be diluted by tissue fluid and decomposed by protease after implantation, so it is difficult to maintain sustained drug concentration or play an effective role in bone induction. OBJECTIVE: To observe the effect of repairing osteoporotic fracture through calcium phosphate cement/fibrin glue composite as the carrier of bone morphogenetic protein 2. METHODS: Fifty-four female Sprague-Dawley rats were selected to remove bilateral ovaries for making osteoporosis models. Three months later, the middle femoral fracture models were made, and then randomized into three groups, with 18 rats in each group. Kirschner wire fixation group was injected nothing. The fracture end was injected with 0.5 mL calcium phosphate cement/fibrin glue (composite group) or calcium phosphate cement/fibrin glue loaded with recombinant human bone morphogenetic protein 2 (loaded group) . At 4 and 12 weeks after fracture, X-ray examination, micro-CT examination, biomechanical three-point bending test and pathological observation were performed. RESULTS AND CONCLUSION: (1) The fracture healing score in the loaded group was higher than that in the other two groups (P < 0.05) . (2) The bone volume fraction, trabecular thickness and number of trabeculae at 4 and 12 weeks in the loaded group were higher than those in the other two groups (P < 0.05) , and the trabecular segregation was lower than that in the other two groups (P < 0.05) . (3) The maximum load and stiffness at 4 and 12 weeks in the loaded group were higher than those in the other two groups (P < 0.05) , and the elastic modulus at 4 weeks after fracture was higher than that in the other two groups (P < 0.05) . (4) Fibrocartilage callus was mainly seen at 4 weeks after fracture in the Kirschner wire fixation group, and the callus was reconstructed into lamellar bone at 12 weeks after fracture with less callus content. Obvious fibrocartilage callus appeared at 4 weeks after fracture in the composite and loaded groups, and the callus was reconstituted into a plate-like bone at 12 weeks. (5) These results imply that the calcium phosphate cement/fibrin glue composite loaded with recombinant human bone morphogenetic protein 2 can promote the healing of osteoporotic fracture and improve bone strength.

Objective:To investigate the clinical effect of the anterolateral femoral perforator flap（ALTP）pedicled with the oblique branch of lateral circumflex femoral artery（LCFA）assisted by CT angiography（CTA）examination for repairing soft tissue defects of limbs.Methods:A retrospective case series study was made on 51 patients with soft tissue defects of limbs treated in Ningbo No.6 Hospital from March 2015 to March 2020，including 31 males and 20 females at age of 26-63 years［（42.0±8.9）years］. The defects were located at the forearm in 15 patients，at the hand in 13，at the lower leg in 15 and at the ankle in 8. The size of defects ranged from 9 cm×6 cm to 18 cm×10 cm，with the size of flaps from 10 cm×6 cm to 20 cm×12 cm. A total of 33 patients were examined with CTA scanning and Doppler ultrasound（CTA group）and 18 patients with Doppler ultrasound（Doppler group）. All patients underwent debridement and negative pressure closed drainage（VSD）at stage I and were repaired by ALTP pedicled with the oblique branch of LCFA at stage II. The diameter and length of the vessel pedicle was recorded in CTA group before operationand in both groups during operation. The time of flap harvesting in both groups was recorded during operation. The survival of the flap in both groups was observed one week after operation. Zhang Hao's scoring standard was applied to evaluate the outcome at the last follow-up.Results:All patients were followed up for 6-12 months［（9.1±1.5）months］. In CTA group，the diameter of LCFA vessel pedicle measured before operation had no significant difference from that during operation（ P>0.05），while the length of LCFA vessel pedicle before operation［（12.3±2.1）cm］was longer than（10.9±2.2）cm during operation（ P<0.05）. The two group showed no significant differences in the diameter and length of LCFA vessel pedicle during operation（ P>0.05）. The time of flap harvesting in CTA group was（38.5±6.2）minutes，significantly shorter than（51.4±8.4）minutes in Doppler group（ P<0.05）. One week after operation，all flaps survived. Two patients developed flap arterial congestion in CTA group，among whom one survived after surgical revision and one with partially necrosis was healed after dressing change. One patient was found with flap arterial congestion with partial necrosis in Doppler group，who was healed after dressing change. There was no significant difference in postoperative flap arterial congestion between the two groups（ P>0.05）. The patients' satisfaction score in CTA group was（8.5±1.5）points at the last follow-up，higher than（7.4±2.0）points in Doppler group（ P<0.05）. Conclusion:For repairing soft tissue defects of limbs，free ALTP pedicled with the oblique branch of LCFA assisted by three- dimensional CT angiography can accurately get the information of perforator，shorten the flap harvesting time，and obtain satisfactory clinical results as compared to Doppler ultrasound.

Objective To explore the effect of lncRNA of growth arrest-specific 5 (lncRNA GAS5) on the radiosensitivity of colon cancer cells by targeting miR-223.Methods The expressions of lncRNA GAS5 in a few of colon cancer cell lines were detected by real-time quantitative PCR (qPCR).The cell lines with low expression level of lncRNA GAS5 were selected for subsequent study.The effect of overexpression of lncRNA GAS5 on the radiosensitivity of colon cancer SW480 cells was detected by cell cloning experiments.The target gene miR-223 of lncRNA GAS5 was predicted and validated by the bioinformatics database starBase and dual luciferase reporter assays.qPCR was used to detect the expression of miR-223 in various colon cancer cell lines and the influence of lncRNA GAS5 overexpression on the expression of miR-223 in SW480 cells.Results Compared with normal human colonic epithelial cells (NCM460),the expressions of lncRNA GAS5 in the colon cancer SW480,LOVO,HT-29 and SW620 cell lines were significantly lower(t =15.25,8.69,14.42,11.62,P ＜ 0.05),with the lowest level in SW480 cells.Both overexpression of lncRNA GAS5 and down-regulation of miR-223 significantly increased the radiosensitivity of colon cancer cells by decreasing cell survival fraction (at 8 Gy,lncRNA GAS5,t =13.51,P ＜ 0.05;anti-miR-223,t =14.93,P ＜ 0.05)and promoting apoptosis (lncRNA GAS5,t =8.30,P ＜ 0.05;anti-miR-223,t =7.32,P ＜ 0.05).Bioinformatics analysis showed that the 3'sequence of lncRNA GAS5 contained the binding sites with miR-223.After overexpression or downregulation of lncRNA GAS5,the expression of miR-223 was enhanced or reduced.Conclusions The lncRNA GAS5 promotes the apoptosis of colon cancer cells and inhibits its survival by targeting miR-223 expression,thereby increases the radiosensitivity of colon cancer cells.

Objective:To investigate the clinical effect of free superficial iliac circumflex artery perforator flap with different arterial anastomosis methods for repairing soft tissue defects of limbs.Methods:A retrospective cohort study was used to analyze the data of 60 patients with soft tissue defects of limbs treated in Ningbo No.6 Hospital from March 2017 to March 2020, including 38 males and 22 females, aged 21-57 years［(35.6±3.1)years］. The wounds were located on the upper limb in 33 patients and the lower limb wound in 27 patients. The defect area was 4.5 cm × 2.0 cm-17.0 cm × 8.0 cm. All patients received debridement and vacuum sealing drainage (VSD) in stage I, and free superficial iliac circumflex artery perforator flap transplantation in stage II. The flap area was 4.9 cm × 2.6 cm-17.0 cm × 8.0 cm. According to the different anastomotic vessels, the superficial iliac circumflex artery was anastomosed end-to- side to the radial artery in 25 patients, the ulnar artery in 8, the dorsalis pedis artery in 18 and the anterior tibial artery in 9. Flap healing and healing time were detected. Texture and appearance of the flap were observed at the last follow-up. The appearance of the donor area was evaluated by Vancouver scar scale (VSS), and the satisfaction rate of appearance of the recipient area was measured by numerical score.Results:All patients were followed up for 5-15 months［(7.1±1.3)points］. All flaps survived successfully after operation, including arterial crisis in 4 patients, among which 2 had flap perforator anastomosed with the radial artery, 1 with the ulnar artery and 1 with the dorsalis pedis artery. There was no significant difference in flap crisis after end-to-side arterial anastomosis in different recipient areas ( P>0.05). For patients with flap perforator anastomosed with the radial artery, the ulnar artery, the dorsalis pedis artery and the anterior tibial artery, the flap healing time was 15(14, 16)days, 15(14, 16)days, 14.5(14，16)days and 14(14，15.5)days, respectively (all P>0.05). The flaps showed sufficient elasticity and soft texture at the last follow-up. For patients with flap perforator anastomosed with the radial artery, the ulnar artery, the dorsalis pedis artery and the anterior tibial artery, the VSS of the donor area was (10.2±1.5)points, (10.9±1.6)points, (9.4±1.5)points and (9.8±1.5)points at the last follow-up (all P>0.05), and the satisfaction rate of appearance of the recipient area was 9 (8, 10)points, 9(8, 9)points, 9(8, 9)points and 8(8, 9.5)points at the last follow-up ( P>0.05), showing that all patients were satisfied with the appearance of the wound. Conclusions:The transplantation of free superficial iliac circumflex artery perforator flap with different arterial anastomosis methods is safe and reliable in repairing soft tissue defects of limbs with advantages of low occurrence of postoperative vascular crisis, good flap survival, no influence on flap healing by different anastomotic vessels, hidden donor area with small scar and satisfactory appearance of the recipient area. It can be used as one of the common vascular repair methods for free tissue flap transplantation.

Objective To analyze tumor immune microenvironment and related mechanisms in liver cancer.Methods We included 10 cases of hepatocellular carcinoma,hepatitis B patients and healthy volunteers from January 2015 to December 2017 in Shanxi Grand Hospital.We first detected the peripheral and local GM-CSF level in each group,detected myeloid-derived suppressor cells (MDSCs) GM-CSF and pathway-related protein expression.from liver cancer,tumor margin and normal liver tissue through flow cytometry and immunohistochemistry,Finally,we transfected the CCR4-NOT transcriptional complex subunit 7 (CNOT7) recombinant plasmid in the hepatoma cell line,and then detected the related protein expression.Results There was no significant difference for peripheral blood GM-CSF level between liver cancer group,hepatitis group and control group (P＞0.05).The level of local GM-CSF was (32.2±8.9) ng/L,which was higher than that of hepatocellular carcinoma (9.7±2.7) ng/L and normal liver tissue (11.6±2.9) ng/L.The difference was statistically significant (P＜0.05).The proportion of MDSCs at the edge of the tumor was (9.9 ±3.6) ％,which was higher than that of liver cancer (4.0± 1.5) ％ and normal liver tissue (6.3±2.3) ％,and the difference was statistically significant (P＜0.05).Immunohistochemistrydata was consistent with previous data.Compared with normal liver tissue,CNOT7 and STAT3 were highly expressed in liver cancer tissues,while STAT1 was lowly expressed.HepG2 human hepatoma cells were selected for transfection.Compared with the empty plasmid group,CNOT7 expression was decreased in the knocking out group at the same time STAT1 expression was increased,STAT3 and GM-CSF expression was decreased.Conclusion In hepatocellular carcinoma,the secretion of GM-CSF increased and the number of MDSCs increased.Knocking out CNOT7 reduced GM-CSF secretion and activate the JAK/STAT signaling pathway.