Objective To analyse the efficacy of three-dimensional conformal radiotherapy(3D-CRT)for hepatic carcinomas.Methods 81 patients with advanced hepatic carcinoma were treated at every turn of 200-400 cGy to a total dose of 4800-5500 cGy.Average-dose was 4580 cGy.Following up time was 225 monthes.Recent effect was evaluated according to WHO tumor evaluation standard.Results 81 patients accomplished the treatment.The local response rate was 82.7 ％(67/81)after treatment with 21 cases CR,46 cases PR.The 1,2 year overall survival rates were 43 ％,26 ％.In the multivariate analysis,the recent effect,the total dose,metastasis,clinical staging,the diameter of the tumors and Child grade presages had statistical significance for overall survival(all P ＜ 0.05).Conclusion 3D-CRT is preferable therapeutic action for primary hepatic carcinomas and can prolong the survival term.

Objective To evaluate the effect of methylprednisolone on hepatic ischemia-reperfusion (I/R) injury in the patients undergoing hepatolobectomy.Methods Sixty ASA physical status Ⅱ or Ⅲ patients,aged 30-64 yr,weighing 45-75 kg,scheduled for elective hepatolobectomy,were randomized to control group or methylprednisolone group (n =30 each).After induction of anesthesia,methylprednisolone 500 mg (in 100 ml of normal saline) was infused intravenously at 5 ml/min before skin incision in group M.Anesthesia was induced with propofol,fentanyl and cisatracurium.The patients were endotracheally intubated and mechanically ventilated.PETCO2 was maintained at 35-45 mmHg.Anesthesia was maintained with 1％-3％ sevoflurane inhalation,remifentanil infusion,and intermittent iv boluses of fentanyl and cisatracurium.MAP was maintained at 70-100 mmHg and HR at 50-90 bpm.At 10 min before induction of anesthesia,and on postoperative day 1,3 and 5,venous blood samples were collected for determination of the plasma levels of alanine aminotransferase (ALT),aspartate amminotransferase (AST),total bilirubin (TBIL),tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6).Results Compared with group C,the plasma levels of ALT,AST and TBIL were significantly decreased on postoperative day l and 3,and the plasma concentrations of TNF-α and IL-6 were decreased on postoperative day 1,3 and 5 in group M.Conclusion Methylprednisolone can reduce hepatic I/R injury in the patients undergoing hepatolobectomy and inhibition of systemic inflammatory responses is involved in the mechanism.

Objective To establish a radioresistant esophageal squamous cancer cell line,and to identify the radioresistant genes and mechanisms.Methods The radioresistant KYSE410-res cell line was established by repeated exposure of cell line KYSE410 to radiation.The proliferation and apoptosis of esophageal squamous cancer cells were evaluated before and after radiation.The changes in gene expression of the esophageal squamous cancer cells after radiation were determined by gene microarray and analyzed by group t test.The genes with significant difference in expression after radiation were validated.Results The KYSE410-res cells had significantly enhanced proliferation and anti-apoptosis than the KYSE410 cells (all P＜0.05).The result of gene microarray showed that compared with the KYSE410 cells,the KYSE410-res cells had the expression of 463 and 251 genes upregulated and downregulated by no less than 4 folds,respectively.Those genes with different expression levels after radiation were mainly responsible for cell proliferation,adhesion,signal transduction,angiogenesis,reactive oxygen metabolism,cell damage repair,and the MAPK/ERK signaling pathway.OAS2 and UBD were key proteins in the network.In the KYSE410-res cells,the expression of HLA-DQBI,MMP1,NCAM1,ZNF521,GPC6,SELENBP1,LCN15,and TFPI-2 genes measured by real-time PCR was consistent with that measured by gene microarray.Conclusions Abnormal activation of the MAPK/ERK signaling pathway,upregulated expression of OAS2 and UBD,downregulated expression of TFPI-2,and upregulated expression of MMPs may play a role in radioresistance of esophageal cancer cells.

Objective To investigate the role of cancer stem cells in radiation resistance of esophageal cancer and its molecular mechanism, and to provide a theoretical basis for radiotherapy for esophageal cancer.Methods Esophageal cancer cell line TE1 was treated with 8 Gy of radiation. Esophageal cancer cell line with resistance to radiation, TE1-res, was established and screened.Cell counting was used to evaluate cell proliferation.Flow cytometry was used to determine the expression of CD44 (high) CD24(-) CD133(+) and apoptosis in cells.The colony formation assay was used to determine the colony-forming rate and cell survival curve.Bisulfite sequencing PCR was used to determine the methylation status of cancer suppressor genes.Comparison of the data was made by group t test or analysis of variance. Results Compared with TE1 cells, TE1-res cells had significantly enhanced proliferation, a significantly higher proportion of CD44( high) CD24(-) CD133(+) cells, and significantly enhanced resistance to apoptosis (mean value 20.84×105 vs.4.46×105/day, P=0.008;(38.0±2.9)%vs.(10.1±1.3)%, P=0.001;mean value 33.23% vs.10.50%, P=0.003 ) .After treatment with 8 Gy of radiation, TE1-res cells had significantly higher colony-forming rate and D0 value than TE1 cells ((14.3±2.6)%vs.(0.9±0.3)%, P=0.011;3.28 vs.2.19 Gy, P=0.125 ) .Moreover, the promoter methylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B was significantly enhanced in TE1-res cells than in TE1 cells ((89.7±4.9)%vs.(5.0±0.5)%, P=0.001;(92.3±4.7)%vs.(10.4±0.7)%, P=0.001;(90.7±3.7)%vs.(7.9±0.4)%, P=0.001;(83.4±5.7)%vs.(17.2±1.2)%, P=0.002;(90.2±
6.7)%vs.(4.4±1.2)%, P=0.002).Conclusions Cancer stem cells play an important role in radiation resistance of esophageal cancer. The resistance to radiation is closely associated with promoter hypermethylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B.

Objective Pemetrexed and β-elemene can inhibit the growth of tumor cells .This study was to investigate the effect of pemetrexed combined with β-elemene on the proliferation and apoptosis of cervical cancer HeLa cells. Methods Cervical cancer HeLa cells were treated with pemetrexed at the concentrations of 38, 76, 152, 228, and 304μg/mL, and at 24 and 48 hours of treatment subjected to MTT for detection of their proliferation .The experiment included four groups , with the cells treated with β-elemene ( 125μg/mL) , pemetrexed ( 76 μg/mL ) , β-elemene ( 125 μg/mL ) +pemetrexed (76μg/mL), and nothing (blank control) for 24 hours, followed by determination of their proliferation and apoptosis by MTT and flow cytometry, respectively. Results Pemetrexed at 38, 76, 152 and 228μg/mL inhibited the proliferation of the HeLa cells in a concentration-dependent manner, with the inhibition rates of (7.24 ±3.78), (7.94 ±4.37), (11.10 ±2.86) and (15.88 ± 3.38)%at 24 hours, and (16.69 ±0.95), (22.54 ±1.53), (24.48 ±0.92) and (25.54 ±3.61)%at 48 hours, both with statis-tically significant differences between any two groups (P<0.05).Significant differences were also found in the proliferation rate of the same concentration of pemetrexed at the two time points (P<0.05).The combination of pemetrexed and β-elemene showed an inhibi-tion rate of (49.95 ±5.76)%at 24 hours, remarkably higher than (24.36 ±5.59)%in theβ-elemene group and (10.69 ±1.37)%in the pemetrexed group (P<0.01). Conclusion Pemetrexed combined with β-elemene can significantly inhibit the proliferation and synergistically accelerate the apoptosis of HeLa cells .

Objective To establish a method based on 3D printing radiology equivalent phantom for individual radiotherapy dose verification,and to offer an assurance for the safety of 3D conformal radiotherapy.Methods Two patients' CT data was collected,reconstructing the first patient's skull and brain tissue to generate a skull-brain phantom for the purpose of testing the equivalent material.The second patient's data was used for whole head tissue reconstruction to produce a head phantom with equivalent material.By inserting ionization chamber dosimeters to target region for radiotherapy program,equivalent phantom dose distribution of lesions location was obtained in order to verify and calibrate the actual radiation treatment planning for patients.Results DR,CT images of the phantoms revealed that the difference of X-ray gray value between brain skull phantom and patient's skull was 13 721,CT value difference between equivalent tissue of brain skull phantom and that part of the patient was 35-40 HU,and CT difference between head phantom temporalis and that of the patient tissue was 18-28 HU.The imaging data indicated that the radiation equivalence of 3D printing phantom was similar to that of human body tissue,and the equivalent dose distribution accorded well with the normal range of treatment.The dose verification of phantom model can effectively improve the accuracy of the radiotherapy system.Conclusions The personalized radiotherapy phantom which based on the 3D printing and tissue equivalent technology is suitable for personalized radiation therapy validation.With advantages of easy accessibility,highly-personalized degree and high precision,this technology provides a reliable and safe way for radiation therapy.

Objective To research the changes in hippocampal voltage-gated sodium channel of Lithium chloride-Pilocarpine epileptic rat models,including Ⅰ sodium channel α subunit protein (Nav1.1),mRNA of Ⅰ sodium channel alpha subunit protein gene and function of sodium channel.Methods Epileptic rat models of Lithium chloride-Pilocarpine were established.Nav1.1 expression in the hippocampus of experimental rats was detected by immunohistochemical staining method,and the changes in voltage-gated sodium channel function (the current-voltage curves,activation and inactivation curves and the recovery curve) of hippocampus nerve cells were detected by whole cell patch-clamp technique.Results (1) The Lithium chloride-Pilocarpine rat models were successfully reproduced.Three stages of behavior (acute,latent and chronic) of rat models were observed.The blank control group was free of seizure.(2) Immunohistochemistry results:neurons in CA1 and DG regions of hippocampal of epileptic rats were normal,and there was no obvious change in the expression of Nav1.1.In CA3 area,the degeneration and necrosis of neurons were obvious.Staining of Nav1.1 became superficial and even disappeared in these areas,but the normal tissues were enhanced around degenerative and necrotic neurons.Compared with the blank control group,the expression of Nav1.1 in the model group was higher(0.235 ±0.008 vs.O.210 ±0.002),and there was statistically significant difference (t'=-7.426,P ＜ 0.05).(3) The whole-cell patch-clamp technique showed that the sodium current density of the model group increased significantly compared with that of the blank group [(-319.70 ± 28.24) pA/pF vs.(-229.06 ± 26.01) pA/pF,t =8.178,P ＜ 0.05],the threshold value of activation curve decreased (4.15 ± 0.80 vs.4.50 ±0.85,t =11.020,P ＜ 0.05),the threshold value of inactivation curve increased (7.47 ± 0.53 vs.6.24 ±0.31,t =6.940,P ＜ 0.05),and the recovery time after inactivation shortened [(1.36 ± 0.15) ms vs.(1.86 ± 0.21)ms,t =6.712,P ＜ 0.05],and there were all statistically significant differences.Conclusion Repeated seizures can lead to increase Nav1.1 compensatory expression of,and significantly increase sodium channel current density,while the threshold value of activation curve decreases,the threshold value of inactivation curve rises,and the recovery time after inactivation is shortened,which eventually leads to increased neuron excitability and is more likely to cause seizures.

Objective: To explore the relationship between genotype and phenotype of ABCB11 deficiency. Methods: Clinical data of two siblings with ABCB11 deficiency were retrospectively analyzed. Related literature from PubMed, CNKI and Wangfang databases was reviewed to date (up to August 2017) with 'ABCB11 gene’ or 'bile salt export pump’, 'cholestasis’ and 'child’ as key words. Results: The patients were siblings. Both of them presented as jaundice, pruritus and hepatosplenomegaly since 3 days after birth. Significant laboratory findings on admission of the older sister included high total bilirubin, 170 µmol/L;conjugated bilirubin, 115.8 µmol/L;alanine aminotransferase, 168 U/L;total bile acid 186.3 µmol/L and normal gamma-glutamyl transpeptidase. While routine laboratory data of the younger brother were as follows: total bilirubin, 148.8 µmol/L;conjugated bilirubin, 96.3 µmol/L;alanine aminotransferase, 232.8 U/L;total bile acid 226 µmol/L, and normal gamma-glutamyl transpeptidase.Both received ursodeoxycholic acid and fat-soluble vitamins. Liver pathology of the younger brother showed giant hepatocytes with ballooning degeneration, focal necrosis and intrahepatic cholestasis. Both the patients harbor the same compound heterozygous mutations in ABCB11 gene, c.145C>T (p.Q49X) and c.1510G>A (p.E504K). The sister is 9 years old now, with normal liver function. Jaundice faded around 3 months after birth, pruritus relieved at age 5, and medications was stopped since then. The brother progressed to liver failure after an operation on perianal abscess when he was 8-month-old, and received living-related liver transplantation when he was 9 month and 20 days old (from his mother). Now he is 1 year and 5 months old, with normal liver function. Both are under our follow-up. Literature review revealed 18 ABCB11 deficiency patients from 7 families who had apparent different prognoses, within each family the siblings had the same ABCB11 gene mutation. Seven cases relieved after ursodeoxycholic acid therapy and/or partial external biliary diversion, 5 received orthotopic liver transplantation, 2 developed hepatocellular carcinoma and 4 cases died in childhood. Conclusions The clinical manifestations of ABCB11 deficiency may vary greatly in patients carrying the same genotype, even in siblings. Patients should be managed in individualized maner.

Objective:To explore the efficacy of digital and 3D printing technologies on design of superficial iliac circumflex artery flap for coverage the donor site of anterolateral thigh flap(ALTF).Methods:Clinical data of 8 patients were studied retrospectively for treatment of soft tissue defects of hand in the Department of Hand Surgery, Wuxi NO.9 People's Hospital Affiliated to Soochow University, from April 2017 to October 2021. The patients were 6 males and 2 females, aged from 29 to 59 years(mean, 45.8 years). Cause of injury: 3 patients were crushed, 2 by hot pressing, and 3 by machine strangulation. Site of injury included: 5 cases were dorsal hand defects and 3 cases were palm defects. All the wounds were contaminated to varying degrees with soft tissue defects. The areas of soft tissue defect ranged from 11 cm×10 cm to 22 cm×14 cm. Four patients had combined injuries of open fracture of metacarpals and phalanges and 3 with tendon defects. All wounds were repaired by free ALTF transplantation. And the donor sites in the thigh were repaired by superficial iliac circumflex artery flaps. The secondary wounds caused by flap harvesting on abdominal wall were closed directly. The targeted perforator vessels were detected preoperatively by CTA combined with CDU. 3D printed models of the affected hand were obtained before operation for individualised repairs according to the shape and area of the wounds. After the operation, all patients entered scheduled follow-ups at the outpatient clinic and via internet by observing the flap shape and testing the recovery of sensory and movement of adjacent joint.Results:The shapes and sizes of the wounds and the flaps were found basically in accordance with those in the preoperative simulative designs. All flaps in 8 patients survived and the wounds healed completely. All patients entered follow-ups for 8 to 24(average, 17.5) months. The donor thighs presented good appearance and colour, pliability without bloating. The range of motion of the hips and knees was not affected. Only linear scars remained in the abdominal donor sites, with natural colour and appearance.Conclusion:Digital and 3D printing technologies in preoperative design of flaps can help to locate the perforator vessels intraoperatively and guide the individualised design of the flaps with improved operation efficiency and satisfactory appearance of the flaps.

Objective:To evaluate the effect of upregulation of HuR gene on the radiosensitivity of esophageal squamous cell carcinoma cell Kyse450. Methods:The HuR gene of Kyse450 cells was upregulated by lentivirus. At the same time, X-ray irradiation at a dose of 6 Gy was selected as the intervention condition. Western blot and qPCR were used to detect the expression levels of protein and RNA after Kyse450 transfection, respectively. CCK8 kit was employed to determine the cell proliferation rate. Clone formation assay was adopted to evaluate the ability of cell clone formation. Wound healing experiment and the Transwell test were performed to detect changes in cell migration. Results:CCK8 assay showed that the proliferation ability of cells was enhanced after upregulation of HuR gene, and this enhancement trend was more obvious after radiation. The plate cloning experiment showed that with the increase of radiation dose, the clone formation rates were decreased in both groups, but the clone formation rate in the overexpression group was higher than that in the control group. Wound healing experiment and Transwell test demonstrated that the wound healing rate and migration ability in the overexpression group were higher than those in the control group, and the difference was more significant after radiotherapy. Western blot showed that the levels of MMP9 and MMP2 at 24 h after radiotherapy in the overexpression group were higher than those in the control group. Conclusion:The upregulation of HuR can enhance the proliferation, cloning, migration capabilities and decrease the radiosensitivity of esophageal squamous cell carcinoma cells.