ObjectiveTo study the effect of TGF-? 1 signal transduction blocked by antisense-Smad 4 gene on progression of experimental liver fibrosis of Kunming mice.MethodsWe transfered retrovirus-mediated antisense Smad 4cDNA into the CCl4/ethanol induced cirrhotic liver of Kunming mouse model to investigate the antisense-Smad 4 gene integration by Southern blot. The expression of Smad 4 in cirrhotic liver was observed by Northern blot, RT-PCR, and Western blot. The fibrotic degree of the livers among the three groups were compared.ResultsThe antisense-Smad 4 gene was confirmed to integrate into the liver of the retrovirus-treated mouse .In the non-treated cirrhotic liver, the expression of Smad 4 mRNA was significantly increased than normal liver, the antisense Smad 4 gene could decrease the expression of Smad 4. Compared with the non-treated liver, the fibrotic septa of the liver in the retrovirus-treated mouse were fewer,narrow, and incomplete,and their fibrotic degree of the treated-liver was reduced.ConclusionThese results indicate that antisense-Smad 4 gene can reduce the expression of Smad 4,inhibit the progression of liver fibrosis.

BACKGROUND: Spinal somatosensory evoked potential(SEP) monitoring is widely used intraoperatively due to the easiness to operate and its reliability.OBJECTIVE: This study was designed to assess the significance of improving SEP signals in intraoperative spinal monitoring on predicting the post-operative spinal function.DESIGN: A non-randomized concurrent controlled study was conducted on selected patients.SETTING: This study was conducted at the Orthopedic Department of Peking Union Medical College Hospital.PARTICIPANTS: Totally 34 patients with cervical spondylosis underwent surgical treatment at the Orthopaedic Department of Peking Union Medical College Hospital were selected from January to October in 2001. Of all the patients, 24 underwent anterior decompression and fusion, 3 single door operation and 7 double door operation. According to the variance of intraoperative SEP, the patients were divided into the improvement group(12 cases)and the non-improvement group(22 cases).METHODS: All the patients' neurologic deficits were assessed according to the Japanese Orthopaedic Association scoring system(JOA score), prior to operation and postoperative week 1, 2, and 4 and month 3, 6. Each patient received the intraoperative spinal SEP monitoring. The variance of SEP signals in amplitude and latency were classified as improvement(an increase in amplitude of 50% or more, or a decrease in latency of 10% or more), decrease(a decrease in amplitude of 50% or more, or an increase in latency of 10% or more), and no improvement.MAIN OUTCOME MEASURES: JOA scores were calculated in two groups in the study at all time points.RESULTS: All the 34 patients entered the statistical analysis procedure. In postoperative week 1 and week 2, the improvement group showed a larger increase in JOA score than the non-improvement group did[improvement group:(14.08±1.44), (14.17±1.11) points; no improvement group:( 12.73 ± 1.42), ( 12.86 ± 1.28)points, P ＜ 0.05]. In postoperative week 4, month 3 and month 6, both groups showed an increase in JOA scores [improvement group: (14.00±1.04), (13.58±1.08), (13.68±1.61)points; no improvement group: (13.82 ± 1.01), (13.41 ± 1.22), (13.41± 1.47)points], but there was no significant difference( P ＞ 0.05).CONCLUSION: Improvement of intraoperative SEP can be used to predict the good early clinical outcomes in surgery for cervical spondylosis.

Based on the body height and weight of the Chinese young men, 975 from the north and 921 from the south of China, actually measured, a calculating formula for the ideal body weight of young men both of north and south listed blow was established. The Northener's ideal body weight (kg) = [height (cm) - 150] ? 0.6 +50; the Southener's ideal body weight(kg) = [height (cm.) - 150] ? 0.6 +48. The formula later checked by another data of body height and weight of 783 young men from north and 883 from south has been proved to be reliable. It also showed that the ideal body weight of 1464 young men from north and 2264 from south calculated according to the formula seemed to be more reasonable than the another's formula used previously.

Objective To investigate the effect of a new type of retrograde reperfusion on the intraoperative internal environment and hemodynamics during classic orthotopic liver transplantation (OLT).Methods 20 patients underwent classic OLT using a new type of retrograde reperfusion in our center.Blood sampling was done at different parts or time points including:before blood venting via the portal vein (PV) of the donor liver,5 ml of blood was collected from the superior vena cava (PSVC),inferior vena cava (PIVC) and portal vein (PPV).During retrograde reperfusion through the inferior vena cava (IVC),5 mL of blood was collected when the volume of blood venting reached 5 ml (T1),100 ml (T2),and 200 ml (T3),respectively.The differences of data were compared after the blood samples were analyzed.In addition,the number instances of postreperfusion syndrome (PRS) were recorded.Results All operations were completed successfully,in which PRS occurred in 4 patients (20％).The most notable findings are the significant changes of nearly all data at T1,T2 and T3,including pH value,PvO2,SvO2,BEecf,HCO3-,Lac,K +,and Ca2 +,compared with PIVC (P ＜ 0.05) and a trend toward recovery in all the data.Yet their levels at T3 did not come back to the levels at PIVC (P ＜ 0.05).Besides,for pH value,Lac,K +,HCO3-and BEecf,there were no significant differences between PSVC,PIVC and PPV (P ＞ 0.05).Conclusions During classic OLT,the main factors leading to a disordered internal environment after recirculation stem from venous retum within the donor liver.This new type of retrograde perfusion can eliminate some of the harmful metabolites inside the donor liver in time and to some extent reduce internal environment disorders as well as drastic hemodynamic fluctuations after recirculation.

Objective To compare health related quality of life (HRQOL) between modified and traditional cutaneous ureterostomy, and explore the reasons for these differences, in order to provide the basis of HRQOL for the choice of cutaneous ureterostomy. Methods A total of 53 patients underwent cutaneous ureterostomy were selected, and the patients were divided into traditional cutaneous ureterostomy group (traditional group, 21 cases) and modified cutaneous ureterostomy group (modified group, 32 cases) according to the surgery method. The patients were evaluated by functional assessment of cancer therapy-bladder (FACT-BL), and the HRQOL was compared between 2 groups. Results There were no statistical differences in HRQOL score at 1, 3, 6 and 9 months after surgery between 2 groups (P>0.05). The HRQOL score at 12 months after surgery was significantly higher in modified group than that in traditional group:(141.5 ± 10.4) scores vs. (123.1 ± 5.2) scores, and there was statistical difference (P<0.01). There were no statistical differences in the scores of physiology status, society/family status, emotional state and functional assessment of cancer therapy-general (FACT-G) at 12 months after surgery between 2 group (P>0.05). But the scores of functional status and bladder cancer special scale (BSS), total score of FACT-BL in modified group were significantly higher than those in traditional group:(26.0 ± 2.5) scores vs. (23.8 ± 3.5) scores, (46.7 ± 6.2) scores vs. (34.8 ± 5.5) scores, (143.9 ± 15.7) scores vs. (117.5 ± 8.1) scores, and there were statistical differences (P<0.01). Conclusions The HRQOL at 12 months after surgery in modified cutaneous ureterostomy is better than that in traditional cutaneous ureterostomy. Therefore, if the patient's physical condition permits, priority should be given to modified cutaneous ureterostomy to reduce the complications and improve the quality of life.

Objective To compare the clinical therapeutic effect of percutaneous nephrolithotomy (PCNL) and flexible ureteroscope lithotripsy (FUL) for unilateral lower-calyceal calculi with the diameter of 10-20 mm. Methods The clinical data of 65 patients with unilateral lower-calyceal calculi with the diameter of 10-20 mm were retrospectively analyzed. Thirty cases were treated with PCNL (PCNL group), and 35 cases were treated with FUL (FUL group). The operative time, success rate of lithotomy, haemoglobin decrease after operation, postoperative hospital stay, hospitalization expenses and complication were compared between 2 groups. Results Treatment was completed successfully in the patients of 2 groups, without ureteral perforation, avulsion and other serious complications intraoperatively and postoperatively. There were no statistical differences in success rate of lithotripsy, incidence of high fever after operation and postoperative analgesia rate between 2 groups (P>0.05). The operative time and hospitalization expenses in FUL group were significantly higher than those in PCNL group:(95.27 ± 22.69) min vs. (62.25 ± 20.73) min and (17 242 ± 2 679) yuan vs. (14 205 ± 1 654) yuan, and the haemoglobin decrease after operation and postoperative hospital stay time were significantly lower than those in PCNL group:(0.67 ± 0.33) g/L vs. (7.98 ± 4.33) g/L and (3.75 ± 0.78) d vs. (6.54 ± 1.68) d, and there were statistical differences (P<0.05). Conclusions For the treatment of lower-calyceal calculi with the diameter of 10-20 mm, the success rates of lithotripsy of PCNL and FUL are similar. FUL has less trauma, with shorter postoperative hospital stay time, but the cost is relatively high.

Sepsis is divided into sepsis, severe sepsis and septic shock according to the severity. The severe sepsis and septic shock can lead to multiple organ dysfunction syndrome, which is life-threatening. In the early stage of sepsis, catabolism is obviously. Early enteral and parenteral nutrition should actively correct nutrients and vitamin deficiency, and provide the best low protein and moderate amount of non-protein calories. In the recovery period, the body needs enough protein and calories. In different periods of sepsis, patients have different nutritional needs. Therefore, mastering the nutritional support treatment for the sepsis is extremely important for the prognosis of patients.

Objective:To identify pathogenicity of the potential splicing variants in two Chinese Han patients with osteogenesis imperfecta.Methods:Genomic DNA was extracted using the conventional phenol-chloroform method; whole exome sequencing (WES) was used to analysis the disease-related variants in the two probands; Minigene assay was used to identify pathogenicity of the variants found in the patients' genome that possibly affect RNA splicing.Results:Two potential splicing variants, c.858+1_858+5delGTAAG in intron 12 of COL1A1 and c.1405-7C>T in intron 24 of COL1A2, were found in proband 1 and proband 2, respectively. In addition, a missense mutation, c.2972G>T (p.G991V) in exon 45 of COL1A2, was detected in proband 2. Minigene assay revealed that the variant in proband 1 caused the skipping of exon 12, while the variant in proband 2 did not lead to aberrant splicing. G199 of the COL1A2 in proband 2 was a highly conserved amino acid site, and the results suggested that c.2972G>T (p.G991V) may be the real pathogenic variant by the means of bioinformatics analysis.Conclusion:The variant c.858+1_858+5delGTAAG in COL1A1 was a causative variant that led to OI in proband 1, while the missense variant c.2972G>T (p.G991V) in COL1A2 was the cause of OI in proband 2, instead of the variant c.1405-7C>T. Minigene assay for potential splicing variants detected by WES could not only validate the pathogenicity of the candidate variants and enrich the mutation spectrum of OI, but also lay the foundation for patients' prenatal diagnosis and subsequent mechanism research.

Objective To investigate the effect of FAM83A on the stem cell-like phenotype, chemosensitivity and radiosensitivity of PANC-1 cells, aiming to provide new ideas for clinical combination therapy of pancreatic cancer. Methods The PANC-1 cells with stable silencing FAM83A were constructed by using lentivirus and validated by qPCR and Western blot. Flow cytometry was used to detect the number of CD133 positive cells and cellular apoptosis; the sphere formation assay was used to test the ability of sphere formation of PANC-1 cells;the effect of gecitabine on the cell viability was detected by MTT assay;the effect of radiation on the proliferation of PANC-1 cells was detected by colony formation assay; the effect of FAM83A on Wnt/β-catenin pathway was examined by Western blot. Results The expressions of FAM83A protein ( 0.83 ± 0.08 ) and mRNA ( 0.29 ± 0.03 ) in PANC-1 cells with stable silencing FAM83A were significantly lower than those in the scrambled control group, respectively (1.95 ± 0.19, 0.98 ± 0.09;t=9.410, 12.600, P<0.05). After silencing FAM83A, the expression of stem cell marker CD133 (8.97 ± 0.62) and the sphere formation ability (8 ± 1) also decreased significantly compared with the scrambled group, respectively (21.60 ± 2.60, 25 ± 3; t=8.184, 9.311, P<0.05), and the stem cell-like phenotype of PANC-1 cells was also significantly inhibited. When PANC-1 cells were silenced by FAM83A and further treated with 50 μmol/L gecitabine at 72 h, the activity of FAM83A-silenced PANC-1 cells (32.33 ± 3.05)％ was significantly lower than that of the gecitabine alone treated group (63.06 ± 5.98)％ (t=6.378, P<0.05), and the apoptosis rate of FAM83A-silenced PANC-1 cells (76.52 ± 8.34) ％ was significantly higher than that of gemcitabine alone group (40.88 ± 4.91)％(t=7.929, P<0.05). After silencing FAM83A combined with IR irradiation, the activity of PANC-1 cells (43.25 ±4.21)％ was significantly lower than that of IR alone (78.13 ± 7.98)％ (t=6.694, P<0.05), and the apoptosis rate (44.56 ± 5.32)％ was significantly increased compared with IR alone (15.15 ±1.95)％ (t = 8.990, P < 0.05). After silencing FAM83A, the expression of Active-β-catenin was significantly decreased while the expression of p-β-catenin was significantly increased, the expression of β-catenin in the nucleus was significantly reduced, although total β-catenin had no significant change, and the activity of Wnt/β-catenin signaling pathway was significantly inhibited. Conclusions Silencing FAM83A could significantly reduce the stem cell-like traits and enhance the chemosensitivity and radiosensitivity of pancreatic cancer cells to gemcitabine and radiation via Wnt/β-catenin signaling pathway, which may provide a new target for targeted and combination therapy of pancreatic cancer.

Objective To study the effect of inhibiting the expression of FOXD1 gene on the radiosensitivity of colorectal cancer cells. Methods The expressions of FOXD1 mRNA and protein in human colorectal cancer tissues and cells were detected by Real-time PCR ( qRT-PCR) and Western blot. The colorectal cancer cell line HCT116 was irradiated with 0, 2, 4 and 6 Gy of X-rays. The expression of FOXD1 in each groups were detected by qRT-PCR and Western blot. HCT116 cells were transfected with FOXD1 siRNA and its negative control and termed as si-FOXD1 group and si-NC group. When these cells were irradiated with 4 Gy X-rays, they were termed as si-FOXD1+4 Gy group and si-NC+4 Gy group. Cell proliferation was detected with MTT method, cell survival fraction was measured with colony formation assay, and DNA-PK activity was detected by TECT DNA-PK kit. The siRNA-transfected colorectal cancer cells were inoculated into BALB/c nude mice to establish the xenograft model. After irradiation, the volume and quality of the subcutaneous transplanted tumors were measured every 5 days. Results The expression of FOXD1 mRNA and protein in colorectal cancer tissues was higher than that in adjacent normal tissues (t=5. 579, 4. 816, P<0. 05). The mRNA(t=5. 85-17. 62, P<0. 05)and protein(t=9. 04-11. 42, P<0. 05) expression of FOXD1 in different colorectal cancer cell lines was higher than that in colonic mucosa epithelial cell line NCM460. The expression of FOXD1 in colorectal cancer cells HCT116 was increased after radiation in a dose dependent manner(t=9. 13-44. 15, P<0. 05). Transfection of si-FOXD1 effectively inhibited the expression of FOXD1 (t=10. 51, P<0. 05), decreased proliferation (t=10. 41, P <0. 05), increased radiosensitivity with a radiosensitization ratio of 1. 797, and reduced the radiation-induced DNA-PK activity ( t = 6. 20, P < 0. 05 ) in colorectal cancer cells. After localized irradiation, the tumor volume and weight in nude mice transplanted with si-FOXD1 HCT116 cells were significantly smaller than those in HCT116 (t=11. 29, 3. 69, P<0. 05). Conclusions Knock-down of FOXD1 gene increases the radiosensitivity of colorectal cancer cells and inhibits the growth of colorectal cancer xenograft in nude mice, which provides a potential target gene in improving the effect of radiotherapy on colorectal cancer.