Objective:To identify the function of the 5′-flank upstream regulation region of human CD226 gene.Methods:The upstream regulation region of CD226 was cloned by PCR and ligated into pGL3 vector. Then the vector was transfected into Jurkat cell and luciferase activity was detected after 48 h culture.Results:CD226 gene may have two promoters, P1 and P2,which were located at the region of -843--319 bp and +1-+181 bp respectively, and PMA can up-regulate P1 while down-regulate P2. Both P1 and P2 can be up-regulated by A23187, especially P2.Conclusion:CD226 gene may have two promoters, and PMA and A23187 can regulate CD226 promoter activity in the similar pattern of protein level.

Objective To explore the application effect of Sandwich teaching method in clinical teaching in the department of neurosurgery.Methods Totally 144 clinical medicine students were divided into 2 groups:Sandwich teaching group (n=72) and traditional teaching group (n=72).The analysis of test and the questionnaire of the students were conducted after the course.Comparison between the two groups was made using independent sample t test and measurement data were expressed as as ± s.P＜0.05 signifies statistically significant differences.Rank sum test was used to compare the distribution of the test scores.Results The test score was statistically higher in Sandwich teaching group than control group [(76.38 ± 10.84) vs.(69.67 ± 10.50)].And the main distribution of the test scores was 70～90 in Sandwich teaching group while 60～80 in control group.Questionnaire showed that the scores of Sandwich teaching group were statistically higher than those ofthe control group in five aspects such as enhancing self-learning ability,improving the ability to analyze and solve problems,and so on.Conclusions Sandwich teaching method achieves good teaching effect and it is worth recommending in clinical teachifig.

Objective To explore the effect and mechanism of Kruppel-associated box(KRAB) type zinc-finger protein Apak on the transcription of ribosomal RNA(rRNA) under hypoxic conditions.Methods Real-time quantitative PCR(qRT-PCR) was used to detect the level of rRNA and the effect of Apak on the transcription of rRNA.The expression and phosphorylation level of proteins were examined by Western blotting.Immunofluorescence assay was used to observe the location of Apak.Results Under hypoxia (0.3%O2),the transcription level of rRNA increased first and then decreased in Apak wild type cells.In Apak knock-out cells,the transcription level of rRNA kept decreasing under hypoxic conditions.The repression of Apak on rRNA synthesis and the nucleolus location of Apak disappeared,and the expression and phosphorylation level of Apak were down-regulated under hypoxic conditions for 3 h.Conclusion Under hypoxic conditions,the disapperance of Apak repression on rRNA transcription could lead to a temporary increase in rRNA.

In order to investigate the impact of fulvic acid (FA) on the hydroxylysyl glycosylation in collagen bio-synthesis, 40 NMRI mice were divided into two groups (n = 20 in each group, consisting 10 females and 10 males). The animal was maintained for two generations by different diets: control group with normal water and food and study group with water containing 30 mg/L FA and normal food. The second generation of the animal was slaughtered, and the biochemical parameters of collagen content and the degree of collagen hydroxylysyl glycosylation in skin, rib and tibia were detected by biochemical methods. The mean value of collagen in the study group was increased slightly, and no significant difference between study group and control group was found (P > 0.05), but the content of glucose-glactose-hydroxylysine (GGH) was significantly decreased in the study group in comparison with the control group (P<0.01). It was suggested that through the decrease of GGH 30 mg/L FA could inhibit the activity of galactosyl-hydroxylysylglucosyl-transferase and further disturb the post-translational modification of collagen intracellularly.
Benzopyrans/*pharmacology ; Bone Development ; Bone and Bones/chemistry ; Bone and Bones/*metabolism ; Collagen/*biosynthesis ; Glycosylation ; Hydroxylysine/*metabolism ; Mice, Inbred Strains ; Osteoarthritis/etiology ; Selenium/deficiency

BACKGROUND:Santoni put forward the cortical bone trajectory technology by changing the traditional pedicle screw placement for lumbar internal fixation in order to obtain better control of the screw and bone in 2009. OBJECTIVE:To analyze biomechanical stability of cortical bone trajectory system in the lumbar fusion. METHODS:Twenty fresh newborn calf L3/4, L5/6 motion segment specimens were obtained, and their ranges of motion were detected under different states, as normal controls. Subsequently, twenty samples were divided into cortical bone trajectory screw group and traditional pedicle screw group, which underwent cortical bone trajectory screw fixation combined with posterior lumbar fusion and traditional pedicle screw fixation combined with posterior lumbar fusion, respectively. Without destruction, ranges of motion were detected under different states in both groups. In the revision group, after the test in the traditional pedicle screw group, screw was withdrawn, and cortical bone trajectory screw was used to detect its range of motion under different states. RESULTS AND CONCLUSION:Ranges of motion at bending to the left and right, anteflexion, posterior extension and axial rotation were significantly lower in the cortical bone trajectory screw group and traditional pedicle screw group than in the normal control group (P<0.05). No significant difference in bending to the left and right, anteflexion, posterior extension and axial rotation was detected between the cortical bone trajectory screw and revision groups and traditional pedicle screw group (P>0.05). These results confirmed that cortical bone trajectory technology combined with posterior lumbar fusion can obtain identical stability as the traditional pedicle screw fixation combined with posterior lumbar fusion. Simultaneously, it is a new choice for revision after traditional pedicle screw fixation.

OBJECTIVE:To evaluate therapeutic efficacy of Xuebijing injection in the treatment of severe craniocerebral trau-ma. METHODS:67 patients with severe craniocerebral trauma selected from neurosurgery department of our hospital were Treat-ment method divided into treatment group(33 cases)and control group(34 cases). Observation group was given conventional treat-ment,i.g. oxygen inhalation,dehydration,nourish cranial nerve,anti-infection. Treatment group On the basis of the control group was given Xuebijing injection 50 ml/time,tid,ivgtt. PT,TT,PLT,FIB,CK,LA, D-D,blood gas index (PaCO2,PaO2, HCO3-)of 2 groups were observed after 7 days of treatment,and prognosis of 2 groups were evaluated after 6 months as well as therapettin efficacy. RESULTS:After treatment,PT,TT,PLT,FIB,CK,LA, D-D,PaCO2,PaO2 and HCO3- of 2 groups were all better than before,and the treatment group was better than the control group,with statistical significance (P<0.05). The rate of good prognosis in treatment group (78.79%) was significantly better than in control group (55.88%) after 6 months of treat-ment,with statistical significance(P<0.05). CONCLUSIONS:Xuebijing injection in the treatment of severe craniocerebral trau-ma can improve coagulation function,blood gas levels and the inflammatory reaction,and is conducive to improve the patient's prognosis.

Objective To explore the influence of G4 cyberknife treatment of large hepatocellular car-cinoma on liver function,and to evaluate its treatment safety.Methods Sixty-three large liver cancer patients treated with routine G4 cyberknife treatment were retrospectively analyzed,and then statistical analysis of the difference in liver function before and after treatment was conducted.Results After G4 cyberknife treatment of 1 2 months,the levels of ALT,ALP,TBIL,PA were respectively 23.00 U /L,1 1 1 .00 U /L,1 3.70 μmol/L, (81 .87 ±1 3.94)%.Compared with the levels before treatment [28.00 U /L,32.00 U /L,1 1 .30 μmol/L, (86.07 ±1 4.07)%],there were no signi-ficant differences found (Z =-1 .677,P =0.094;Z =-0.504, P =0.61 4;Z =-1 .945,P =0.053;t =1 .271 ,P =0.21 3).The level of ambumin was (34.84 ±4.75)g/L at 1 2 months after treatment,which decreased and the difference compared with the level before treatment [(37.45 ±4.1 4)g/L]was significant (t =3.357,P =0.002).The Child-Pugh grade was 5.80 ±1 .1 7 respectively at the time points of 1 2 months after treatment,and no significant difference was found compared with the Child-Pugh grade before treatment (5.48 ±0.81 ,t =-1 .668,P =0.1 06).Conclusion G4 cyberknife treatment does not cause liver injury.It is safe and reliable in large liver cancer treatment.So,it is worth widely clinical popularizing.

Objective:This study aimed to establish an eXtreme Gradient Boosting(XGBoost) model that can predict the recurrence of hepatocellular carcinoma(HCC)patients after laparoscopic hepatectomy (LH) surgery.Methods:A total of 440 patients with primary HCC who received LH treatment for the first time from January 2013 to September 2016 in Affiliated Hospital of Chengde Medical University were selected as the research objects. The diagnosis method was pathological diagnosis. Research objects were divided into training group ( n=88) and verification group ( n=352) at a ratio of 2∶8 by random number table method. The Kaplan-Meier method was used to draw the recurrence-free survival curve, and the Log-rank test was used to compare the survival of the two groups; the training group was used to establish the COX regression model and the XGBoost model to screen independent predictors of recurrence after LH; receiver operating characteristic(ROC) curve was used to analyze the predictive abilities of the two models, and conducted internal verification in the verification group; Hosmer and Lemeshow Test was used to evaluate the calibration of the two models, and P>0.05 was used as a good fit between the model and the actual situation. Results:Both the COX regression model and the XGBoost model screened out tumor thrombus, low degree of differentiation, tumor microvascular infiltration (MVI), number of tumors, large tumors, and positive hepatitis B surface antigen were independent predictors of tumor recurrence( HR=2.477, 0.769, 1.786, 1.905, 1.544, 1.805; 95% CI: 1.465-4.251, 0.619-0.819, 1.263-2.546, 1.354-2.704, 1.272-1.816, 1.055-2.555). The XGboost model scores were 32 points, 29 points, 24 points, 18 points, 16 points, 11 points, respectively. In the training group, the area under the curve (AUC) of the COX regression model and XGBoost model for predicting recurrence were 0.746 (0.730-0.762) and 0.802 (0.785-0.818), respectively. The XGBoost model had strong predictive ability and was confirmed in the validation cohort. Conclusions:This study had established and verified the XGBoost model that can predict the recurrence of HCC patients after receiving LH for the first time. It can be used in clinics to assist doctors in formulating personalized postoperative monitoring programs for patients. Early detection, early diagnosis and early treatment of tumors and strengthening of postoperative follow-up are important measures to improve the prognosis of patients.

ObjectiveTo investigate the characteristics of EGFR gene mutations among NSCLC patients in south of China and analyze the correlation between mutations and clinical features.Methods Specimens of lung cancer tissues were collected from 185 NSCIC patients in our hospital.DNA was extracted from specimens.Exon 18,19,20 and 21 of EGFR gene were amplified by FQ-PCR to be bi-directional sequenced.ResultsEGFR gene mutations in 62 (33.5％) of 185 NSCLC patients was identified in carcinoma tissues,of which,2cases,41cases,5 cases and 14 cases respectively located at exon 18,exon 19,exon 20 and exon 21.The mutation of Del L747 → P752 (P753S) ( proportion 8.1％ ),Del E746 → A750 ( proportion 45.1％ ) at exon 19 and L858R ( proportion 22.6％ ) at exon 21 were the predominant mutation in 16 kinds of mutations.Four cases of mutation at exon 19 got the different results in bi-directional sequencing.The silent mutation 2361G→A at exon 20 was observed (28.1％ ).The mutation rate in women was significantly higher than men (46.2％ vs 24.3％,x2 =9.670,P =0.002).Non-smokers had significantly higher mutation rate than smokers (41.4％ vs 17.1％,x2 =7.380,P =0.007) ; Adenocarcinoma patients had significantly higher mutation rate than squamous cell carcinoma (38.3％ vs 6.3％,x2 =6.426,P =0.011).Clinical stage Ⅲ patients had significantly lower mutation ratethan patients with stage Ⅱ orⅣ ( 10.8％ vs 53.8％,x2 =8.026,P =0.003 ;10.8％ vs 41.3％,x2 =9.518,P =0.002).No statistically significance correlation was found between the mutation ratio and age.ConclusionsEGFR gene mutation has a close relationship with females,non-smokers and adenocarcinoma.Most mutations occur in exon 19 and 20 among patients in south of China.

Objective To investigate the expression variation of aquaporin in colon tissues in acute necrotizing pancreatitis (ANP).Methods ANP rat model was induced by the retrograde injection of sodium taurocholate into the biliopancreatic duct.The rats were killed at 4 h, 8 h, 12 h and 24 h after modeling with 6 rats for each time point.The pancreas and colon tissues were harvested for pathological examination.The levels of IL-6, TNF-α mRNA expression and AQR (aquaporin-3, aquaporin-4, aquaporin-8) mRNA expression in proximal and distant colon were detected by RT-PCR.The levels of aquaporin protein in colon were examined by immunohistochemistry.Results After the establishment of ANP SD rat model, the integrity of colonic mucosa was continuously damaged, the structure of epithelial cells was unclear and the colonic villus were broken and destroyed, and inflammatory cell infiltration in submucosa was observed.The pathological score increased with the time of modeling.In 4 h, except that the mRNA levels of AQP-4 in distal colon was not obviously changed, mRNA levels of IL-6 and TNF-α, mRNA and protein expression of AQP-3 and AQP-8 in the proximal and distal colon of ANP rats were significantly elevated compared with shame group (P<0.05).AQP-3 and AQP-8 mRNA in proximal colon of ANP rats reached its peak in 8 h after the establishment and AQP-4 mRNA peaked at 24 h.AQP-3 and AQP-4 mRNA in distant colon of ANP rats reached its peak in 8 h after the establishment and AQP-8 mRNA peaked at 24 h.Protein expression of AQP-3, AQP-4 and AQP-8 in proximal and distant colon was strongest in 12 h and 24 h after the establishment.Conclusions With the progression of the ANP, the expression levels of AQP-3, AQP-4 and AQP-8 in both proximal and distal colons were elevated in various degrees, indicating that the aquaporins may participate in water metabolism of colon during ANP.