OBJECTIVE To construct siRNA eukaryotic expression vector targeting protein kinase CK2?and to investigate its inhibitory effect on invasion of the HEp-2 cell line in human laryngeal carcinoma. METHODS siRNA expression vector psiRNA-hH1neo-CK2 targeting protein kinase CK2?was constructed by gene recombination,and then was transfected into the HEp-2 cells by lipofectamine methods. Protein kinase CK2?mRNA and protein of the transfected cells were detected by reverse transcription polymerase chain reaction(RT-PCR) and Western blot,respectively. The invasion of the transfected cells was measured by Boyden chamber.SP method was used to examine the expressions of MMP2 and TIMP2 protein of the transfected HEp-2 cells. RESULTS Protein kinase CK2?siRNA expression vector was successfully constructed by gene recombination. Compared with non-specific interfering groups and blank groups, protein kinase CK2?mRNA and protein were significantly decreased respectively in the psiRNA-hHIneo-CK2 groups(P

Objective To evaluate the diagnostic value of three nuclide imaging methods in patients with hyperparathyroidism. Methods Thirty five patients with hyperparathyroidism underwent 201 Tl/ 99m TcO-4 (8 cases), double phase 99m Tc MIBI imaging methods (27 cases) and 99m Tc MIBI/ 99m TcO-4 substraction imaging (20 cases). Abnormal increase of radioactivity in substraction imaging or delay imaging denoted positive result. All data of nuclide imaging were evaluated according to final clinical results and were compared with ultrasound or CT. Results 35 cases of hyperparathyroidism were proved, including 31 adenomas (ectopic 1), 3 hyperplasia and 1 carcinoma. The sensitivity of 201 Tl/ 99m TcO-4 , double phase 99m Tc MIBI imaging and 99m Tc MIBI/ 99m TcO-4 substraction imaging was 62.5%, 88.9%,90.0%, respectively. The specificity of 3 nuclide imaging methods was 100%. The sensitivity and specificity of ultrasound were 74.3%, 85.7%, respectively. The sensitivity of CT was 78.6%. The results showed that 99m Tc MIBI/ 99m TcO-4 was superior to other imaging. Conclusion An accurate diagnosis of hyperparathyroidism and preoperative anatomic localization can be determined by means of nuclide imaging.

In order to study the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in human laryngeal squamous cell carcinoma (LSCC) and its significance, the expression of VEGF mRNA and COX-2 mRNA in 62 cases of LSCC and 54 adjacent noncancerous laryngeal tissues and 9 normal human laryngeal mucous tissues was detected by using techniques of semi-quantitative RT-PCR. It was found that the expression level of VEGF and COX-2 mRNA was significantly increased in LSCC as compared with that in the normal human laryngeal mucous tissues (both P < 0.01), and the expression level of VEGF and COX-2 mRNA were significantly increased in stage Ill + IV tissues of LSCC as compared with the stage I + II tissues of LSCC (P < 0.01). There was a high positive correlation between VEGF and COX-2 expression in LSCC (r = 0.756, P < 0.01). These data raise the possibility that VEGF and COX-2 may play key roles in the growth, invasion and metastasis of LSCC.
Carcinoma, Squamous Cell/*metabolism ; Cyclooxygenase 2/*biosynthesis ; Cyclooxygenase 2/genetics ; Laryngeal Neoplasms/*metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Tumor Markers, Biological ; Vascular Endothelial Growth Factor A/*biosynthesis ; Vascular Endothelial Growth Factor A/genetics

Objective To investigate the significance of serum neuron specific enolase (NSE) in the severity and prognosis assessments of diffuse axonal injury (DAI).Methods The levels of serum NSE were measured by radioimmunoassay (RIA) at 12 hours and 1st,2nd,3rd,7th and 14th days after injury in 79patients with DAI.The relationship of serum NSE level with the severity and the prognosis of DAI were analyzed in the patients with DAI.Another 15 patients with only limb fracture and without hemorrhagic shock treated in the hospital during the same period served as the control group.Results The serum NSE levels of the mild injury group were (10.47 ± 2.75) ng/L,(13.41 ± 3.45) ng/L,(16.41 ±4.14) ng/L,(15.57 ±4.28) ng/L,(7.95 ±2.79) ng/L,and (6.39 ± 1.55)ng/L at 12 hours and 1st,2nd,3rd,7th and 14th days after injury.The serum NSE levels of the moderate injury group were (14.98 ± 3.78) ng/L,(19.88 ± 4.78)ng/L,(22.41 ±5.50) ng/L,(20.11 ±6.60) ng/L,(14.59 ±6.64) ng/L,and (8.31 ±3.83) ng/L respectively at 12 hours and 1st,2nd,3rd,7th and 14th days after injury.While the serum NSE levels of the severe injury group were (27.22 ± 4.54) ng/L,(36.43 ± 10.38) ng/L,(41.32 ± 12.44) ng/L,(43.98 ±9.51) ng/L,(42.22 ± 13.05) ng/L,and (37.59 ± 12.96) ng/L at 12 hours and 1st,2nd,3rd,7th and 14th days after injury respectively.The NSE levels in each time point were significantly higher in the severe injury group than in the mild and moderate injury groups (F within =28.11,P ＜ 0.001 ; F between =57.34,P ＜0.001 ;F interaction =8.21,P ＜ 0.001 ;P ＜ 0.01).Compared with the control group ((6.26 ± 1.35) ng/L),the serum NSE levels of the DAI group were significantly different at 12 hours after injury ((18.16 ± 3.76)ng/L,t =2.938,P ＜ 0.01).At three months after injury,patients were divided into the decreased group (n =9),poor prognoses group (in vegetative state or severely disabled,n =29) and good prognoses group (moderately disabled or completely recovered,n =41) according to the GOS score.The serum NSE levels of the decreased group were (32.07 ± 5.73) ng/L,(43.12 ± 15.04) ng/L,(48.26 ± 14.89) ng/L,(50.47 ±11.05) ng/L,(52.90 ±3.82) ng/L,and (56.17 ± 14.62) ng/L respectively at 12 hours and 1st,2nd,3rd,7th and 14th days after injury.The serum NSE levels of the poor prognoses group were (21.90 ± 4.95) ng/L,(24.13 ± 9.94) ng/L,(26.43 ± 6.99) ng/L,(21.62 ± 9.77) ng/L,(15.80 ± 7.15) ng/L,and (10.16 ± 2.33) ng/L respectively at 12 hours and 1st,2nd,3rd,7th and 14th days after injury.The serum NSE levels of the good prognoses group were (13.61 ±4.56) ng/L,(13.75 ±5.10) ng/L,(14.77 ±5.41) ng/L,(13.47 ±4.49) ng/L,(8.92 ± 5.61) ng/L,and (6.60 ± 2.30) ng/L at 12 hours and 1 st,2nd,3rd,7th and 14th days after injury respectively.At each time point,the serum NSE levels were significantly different in the decreased group than in the good prognoses and the poor prognoses groups (F within =18.70,P ＜ 0.001 ; F between =62.97,P ＜0.001 ;F interaction =11.83,P ＜0.001).Conclusion The serum NSE levels can be regard as an index for judging the injury severity and prognosis of DAI,and can be used to guide the option and adjustment of therapeutic approaches for patients with DAI.

BACKGROUND:At present, liver transplantation is the best method to treat end-stage liver disease. UW solution is recognized as the best liver preservation solution, but it is expensive. Moreover, the extracel ular fluid of high K+condition is inconsistent with human physiology. Because transient hyperkalemia of UW solution often causes cardiac arrest, research and development of the new liver preservation solution already brook no delay. OBJECTIVE:To study the protective effect of self-designed KYL solution on ischemia reperfusion injury in macaque donor liver. METHODS:A total of eight recipient macaques and eight donor macaques were selected in this study. Each group contained KYL solution group (n=4) and UW solution group (n=4). Donor liver was perfused and cryopreserved for 4 hours and subjected to al ogenic orthotopic liver transplantation. At 30 minutes and 6 hours after transplantation, bile production was recorded. Blood was obtained and used to detect concentrations of aspartate aminotransferase, alanine aminotransferase, nitric oxide, endothelin-1 and tumor necrosis factor-α. Liver tissue was col ected and detected under the light microscope. RESULTS AND CONCLUSION:Bile secretion was found in both groups. Bile secretion production increased as time went on (P<0.05). At 30 minutes and 6 hours after donor liver reperfusion, serum aspartate aminotransferase and alanine aminotransferase concentrations were lower in the KYL solution group than in the UW solution group (P<0.05). No significant difference was found in levels of serum nitric oxide, endothelin 1 and tumor necrosis factor alpha between the two groups (P>0.05). Under light microscope, morphological observation of liver tissue revealed that cel ular edema was evident in the UW solution group than in the KYL solution group. Results suggest that the effect of KYL solution on preventing ischemia/reperfusion injury was identical to the UW solution, and partial effect was better than UW solution.

A new caffeate compound, (E)-erythro-syringylglyceryl caffeate (1), was isolated from the roots and rhizomes of Nardostachys chinensis Batal., together with nine known phenolic compounds, including (+)-licarin A (2), naringenin 4', 7-dimethyl ether (3), pinoresinol-4-O-β-D-glucoside (4), caraphenol A (5), Z-miyabenol C (6), protocatechuic acid (7), caffeic acid (8), gallic acid (9) and vanillic acid (10). Their chemical structures were elucidated on the basis of spectroscopic data and physicochemical properties. Furthermore, this is the first report of compounds 2, 5 and 6 from Nardostachys genus.

BACKGROUND:Acelular dermal matrix is a cel-free natural tissue scaffold similar to human soft tissue, which is easy to shape and has non-toxic side effects. It has been used to repair the urethra and ureter. OBJECTIVE:To investigate the effect of acelular dermal matrix on the repair of bile duct injury. METHODS:Thirty Diannan miniature pigs were randomly divided into three groups: in blank group, the bile duct was resected folowed by end to end anastomosis; in experimental group, bile duct defect model was made folowed by repair with acelular dermal matrix; in control group, bile duct defect model was made folowed by repair with expanded polytetrafluoroethylene. At 6 and 24 weeks after repair, bile duct patches and surrounding tissues were taken for immunohistochemical observation and RT-PCR detection. RESULTS AND CONCLUSION: Compared with the control and blank group, the expression of cytokeratin was higher, but the expression of transforming growth factor β1 was lower in the experimental group. Within 24 weeks after repair, the total mRNA level of transforming growth factor β1 was lower in the experimental group than the other two groups (P < 0.05), but the total mRNA levels of insulin-like growth factor 2 and vascular endothelial growth factor were higher in the experimental group (P < 0.05). These findings indicate that the acelular dermal matrix for repair of bile duct injury can promote angiogenesis and bile duct epithelial regeneration, but not increase the formation of scars.

Objective To explore the clinical efficacy of internal fixation with dynamic anterior plate-screw system for quadrilateral area(DAPSQ) in the treatment of complex acetabular fractures involving the quadrilateral surface.Methods Between January 2005 and January 2017,a series of 143 patients with complex quadrilateral surface fracture of the acetabulum were treated at Department of Orthopaedics,General Hospital of Central Theater Command.They were 86 males 57 females,with a mean age 46.6 years (range,from 19 to 77 years).By the Letournel-Judet classification,there were 38 anterior column plus posterior hemitransverse fractures,26 T-shaped fractures and 79 double column fractures.They were all closed.The delay from injury to surgery averaged 9.6 days(range,from 3 to 21 days).All the cases were treated with DAPSQ internal fixation via the ilioinguinal approach.Their operation time,intraoperative bleeding volume,quality of fracture reduction,hip function and complications were recorded.Results The operation time for this series ranged from 2.5 to 5.0 hours,averaging 3.8 hours.The intraoperative blood bleeding ranged from 400 to 2,500 mL,with an average of 680 mL.By the Matta criteria for fracture reduction,76 cases were rated as excellent,51 as good and 16 as poor,giving an excellent and good rate of 88.8％.The 143 patients were followed up from 4 months to 10 years (mean,4.1 years).The clinical fracture union was achieved after 2 to 4 months (mean,3 months).By the modified Merle d'Aubigné & Postel criteria,the affected hips scored 9 to 18 points (mean,16.3 points) at the final follow-ups,with 64 excellent,52 good,19 fair and 8 poor cases,giving an excellent and good rate of 81.1％.The postoperative complications included lesion of lateral femoral cutaneous nerve in 7 cases,urinary tract infection in 15 ones,traumatic osteoarthritis in 20 ones 5 of whom had to receive total hip arthroplasty,and heterotopic ossification (Brooker Grade Ⅰ) in 5 ones.Conclusion Internal fixation with DAPSQ is safe and effective for the treatment of complex acetabular fractures involving the quadrilateral surface.

Objective To study the potency of transdifferentiated renal tubular epithelial cells for acquiring the tissue stem cells during renal fibrosis.Methods The in vitro cellular model of renal tubular epithelial cells(NRK-52E)transdifferentiation under the inflammatory environment of the local renin-angiotensin (AngⅡ)system was established.The expression and change situation of the embryonic kidney developmental gene Pax2 and the tissue stem cell surface marker CD133 were observed.Results Local high concentration of AngⅡcould stimulate the NRK-52E cells to express Pax2 and CD133 molecule,its effect demonstrated the dose-and time-dependent relation.Conclusion The inflammatory damage leads to the transdifferentiated renal tubular epithelial cells po-tency to acquire the tissue stem cell.

In order to study the effect of 5, 6-Dichloro-1-beta-D-ribofuranosyl-benzimidazole (DRB) on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry (FCM) and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose-and time-dependent manner. After being treated with 0, 10, 20, 40, 80 microm mol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was (0.68+/-0.19)%, (1.95+/-0.12)%, (8.51+/-0.26)%, (11.26+/-0.17)% and (14.99+/-0.32)%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 microm mol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.