Objective To construct the transmembrane TNF-? eukaryotic expression vector in n-3 PUFA dependant manner. Methods PPRE-tk sequence was designed and artificially synthesized, and then inserted it into pcDNA3-TNF( ? 1-12)WB plasmid to construct eukaryotic expression vector pPPRE-tk-tmTNF-? by gene recombine techniques. The tmTNF-? protein expression level was observed in MCF-7 transfected cells incubated with EPA by immunofluorescence technique. Results pPPRE-tk-tmTNF-? expression vector was constructed successfully and identified by agarose gel electrophoretic analysis and nucleotide sequence analysis. EPA could increase tmTNF expression levels in time- and dose-dependant manners. Conclusion tmTNF-? expression vector regulated by n-3 PUFA is successfully constructed.

Objective To study the relationship between plasma free fatty acids composition and the incidence of nonalcoholic fatty liver disease(NAFLD) .Methods By the design of case‐control study ,105 patients with NAFLD as cases and 110 healthy peo‐ple as controls were enrolled into the study .Plasma free fatty acid levels were determined by gas chromatography .Results High level of plasma palmitic acid(C16 :0)(OR=1 .769) was the risk factors of NAFLD ,while plasma levels of linoleic acid(C18 :2 n‐6) (OR=0 .855) and arachidonic acid(C20 :4 n‐6)(OR=0 .181)were negatively associated with the incidence of NAFLD .Conclusion These findings suggest that a proper ratio of diet fatty acids intake may reduce the risk of NAFLD .

Objective:The present study aimed to investigate the short-term efficacy and adverse effects of induction chrono-che-motherapy including docetaxe1 (TXT), cisplatin (DDP), and 5 fluorouraci1 (5-FU) followed by concomitant chemoradiotherapy in lo-co-regionally advanced nasopharyngeal carcinoma (NPC). Methods:Newly diagnosed locally advanced (Ⅲ~Ⅳb) NPC patients were enrolled in this study. All patients received three cycles of TPF regimen. The TPF chemotherapy regimen was administered as follows:TXT, 75 mg/m2, i.v. infusion, d1; DDP, 75 mg/m2, bolus infusion from 10:00 to 22:00, d1-5; and 5-FU 750 mg/m2/d bolus infusion from 22:00 to 10:00, d1-5, with 21 days each cycle, followed by concomitant IMRT and chemotherapy (paclitaxel 135 mg/m2 i.v. infu-sion, with 21 days each cycle and a total of 2 courses). Acute and late toxicities were graded according to the Common Terminology Cri-teria for Adverse Events v3.0 scoring criteria. Tumor response was evaluated using 2000 Response Evaluation Criteria in Solid Tumors criteria. Results:The CR and PR rates of induction chemotherapy were 23.8%and 68.6%, respectively;whereas the CR and PR rates of the combined modality treatment were 64.8%and 31.4%, respectively. Two-year overall survival rate was 91.4%, two-year progres-sion free survival rate was 87.0%, and two-year distant metastasis-free survival rate was 88.4%. The main side effects from induction chemotherapy include an over grade 3 granulocytopenia of 28.6%. Major toxicity from concurrent chemo-radiotherapy was oral mucosi-tis (81.0%);grade 3 to 4 oral mucositis was 16%. No treatment-related deaths occurred in this study. Conclusion:Induction chrono-che-motherapy using TPF followed by concurrent chemoradiotherapy of paclitaxel is a well-tolerated treatment with short-term efficacy and severity for locally advanced NPC. Further follow-up is required to assess the late effects and long-term efficacy.

Objective To assess the series of special anti-fatigue foods including JuntiⅠ, JuntiⅡ, JuntiⅢ, and Jun-tiⅣon military physical performance capacity .Methods Fifty-four soldiers , selected from a border defense troop , were randomly designated to control group , trial group 1 and trial group 2.Subjects of two trial groups were supplied with No .1 nutritional package ( including JuntiⅠ, Ⅲand Ⅳ) and No.2 nutritional package ( including Junti Ⅱ, Ⅲand Ⅳ),re-spectively, while no additional nutritional supplements were added in control group .After 7 days’ supplementation, a hard military exercise was performed to induce fatigue and an increasing load test was used to assess physical activity .RPE scale, exhaustive time and time taken to reach the 75% maximal heart rate were recorded while serum markers , such as glucose, lactate, BUN, LDH,and CK, were detected after test .Moreover, serum lactate and fatigue recovery scale were determined on the evening of the same day and the next morning .Results Prolonged exhaustive time and time taken to reach the 75%maximal heart rate and elevated RPE scores at 6 min were detected in both two trial groups compared with the control group .Meanwhile , after the increasing load test , elevated glucose concentration and reduced lactate , BUN, LDH and CK were also observed in both trial groups .Moreover, serum lactate of both trial groups was quickly recovered on the evening of the same day compared with the control group , and the next morning , serum lactate was even much lower in trial groups than in control group .The fatigue recovery scores were higher in trial groups at both time points .Meanwhile, there was no difference of such indexes between the two trial groups .Conclusion Through the combination use , the series of special anti-fatigue foods, inclucling No.1 and No.2 nutritional packages , can significantly improve the soldiers′physi-cal performance capacity , delay the physical fatigue emergence , promote physical activity recovery and prevent military training injury.

Objective To investigate the structure-activity relationship for 21 anthocyanins in inhibiting oxidized injury of endothelial cells,and explore the structural characteristics of anthocyanins closely related to their effects. Methods Endothelial cells were treated by ox-LDL at different concentrations of 50,100,150 or 200 ?g/ml,and MTT assay was used to determine IC50. After pre-incubated for 2 h with different concentrations ( 50,100 or 200 ?mol/L) of anthocyanins and then treated with 100 ?g/ml ox-LDL for another 24 h in endothelial cells,MTT assay was used to detect the cellular viability. After pre-treated for 2 h with different anthocyanins with 100 ?mol/L and treated with ox-LDL for another 24 h,MDA and NO level in the culture media were both measured according to the methods of assay kits. Structure-activity relationship was analyzed according to the respective cellular viability,MDA and NO level. Results Cellular viability was significantly inhibited by ox-LDL in a dose-dependent manner,and the IC50 was 100 ?g/ml. A significant correlation was observed among the effect of anthocyanins on cell viability,MDA production and NO release. The inhibitory effects of anthocyanins in ox-LDL-injured endothelial cells were positively related to the total number of hydroxyl groups and hydroxyl substitutions in B ring. 3′,4′-ortho-dihydroxyl substitution on B-ring and a 3-hydroxyl group on C-ring significantly enhanced the inhibitory effect of anthocyanins,yet methoxylation or glycosylation significantly decreased the effect. 6-hydroxylation substitution might attenuate the inhibitory effect of anthocyanins,while substitution at C5 or C5′ showed no significant influence on the effect of anthocyanins. Anthocyanin with monosaccharose substitution was much stronger than that with disaccharose substitution,while there was no significant difference between anthocyanins with glucoside and that with galacotoside substitution. Delphinidin and delphinidin-3-glucoside were respectively the most effectual anthocyanidin or anthocyanin. Conclusion 3′,4′-ortho-dihydroxyl substitution on B-ring and a 3-hydroxyl group on C-ring are the main structural requirements for anthocyanins in suppressing ox-LDL-induced injury in endothelial cells.

Objective:To investigate the outcomes of the regimen with docetaxel, cisplatin, and 5-fluorouracil (TPF regimen) in chrono-chemotherapy, and evaluate the feasibility of reducing the toxicity and immunological damage in nasopharyngeal carcinoma (NPC) patients with distant metastasis at preliminary diagnosis, then to compare the advantages and disadvantages between chrono-che-motherapy and traditional chemotherapy. Methods:A total of 46 NPC patients with distant metastasis at preliminary diagnosis (UICC 2010 stage IVc) were enrolled in this study. These NPC patients were randomly divided into chrono-chemotherapy and conventional chemotherapy groups, with 23 cases for each group. TPF neo-adjuvant chemotherapy was conducted in both groups for two cycles, with 21 days to 28 days for each cycle. The following regimen was used for the chrono-chemotherapy group:docetaxel 75 mg/m2, infu-sion, d1;cisplatin 75 mg/m2, 10:00 a.m.-10:00 p.m., continuous infusion, d1-d5;and fluorouracil 750 mg/(m2 · d), 10:00 p.m.-10:00 a. m., continuous intravenous infusion, d1-d5. The following regimen was used for the conventional chemotherapy group:docetaxel 75 mg/m2, infusion, d1;cisplatin 75 mg/m2, infusion, d1;and fluorouracil 750 mg/(m2· d), continuous infusion, d1-d5, 120 h. Patients who obtained therapeutic efficacy via induction chemotherapy were provided with intensity-modulated radiotherapy as a concurrent radio-therapy and chemotherapy (DDP 100 mg/m2, infusion, d1-d2, with 21 days each cycle and a total of two courses). One month after con-current chemoradiation, an adjuvant chemotherapy with the same regimen as the induction chemotherapy was employed for a total of two courses. Acute and late toxicities were graded in accordance with the Common Terminology Criteria for Adverse Events v3.0 scor-ing. Tumor response was evaluated using the 2000 Response Evaluation Criteria in Solid Tumors. The effective rates included complete and partial responses. Relevant data were analyzed by SPSS16.0 statistical software. Results:More emesis was observed at Grade 2 or above in the conventional chemotherapy group than in the chrono-chemotherapy group, with statistical significance between the two groups (P=0.035). After chemotherapy, the value of CD4/CD8 increased in the chrono-chemotherapy group and decreased in the con-ventional chemotherapy group, with statistical significance between the two groups (P=0.033). Conclusion:The proposed chrono-che-motherapy outperforms conventional chemotherapy in reducing the occurrence of severe vomiting. This chrono-chemotherapy may be advantageous in reducing severe bone marrow depression and may play a positive role in the immune function of NPC patients.

Objective To assess the antiangiogenic role of recombinant human endostatin combined with chemoradiotherapy and the capacity,and to explore the early tumor response as measured by comparing the change of MRI perfusion parameter.Methods From May 2012 to March 2013,22 locally advanced nasopharyngeal carcinoma patients who received recombinant human endostatin combined with chemoradiotherapy following induction chemotherapy,were included in the prospective study group.The other 25 patients,who received chemoradiotherapy following induction chemotherapy alone in the same period,were included in the control group.The perfusion parameters including blood volume(BV),blood flux(BF),mean transit time (MTT) were obtained by carrying out MR perfusion scanning at 3 time points:before induction chemotherapy,after induction chemotherapy,the end of concurrent chemoradiotherapy.Results Compared with before induction chemotherapy,the perfusion parameters including BV and BF obviously decreased in the study group (F =3.05,3.85,P ＜ 0.05).The parameter of MTT had no obviously change in the study group(P ＞0.05).In the control group,the change of BV,BF and MTT of nasopharyngeal lesions area during the treatment showed no significant difference (P ＞ 0.05).To make comparison between the two groups,at the end of concurrent chemoradiotherapy,BF of nasopharyngeal lesions area in the study group was 0.72 ± 0.56 and 1.92 ± 1.26 in the control group,the former showing significantly declined results (t =-3.056,P =0.012).Conclusions Recombinant human endostatin might be a good indicator of local tumor microvascular changes and the treatment-related toxicity could be tolerated.Magnetic resonance perfusion imaging maybe assessed the capacity of anti-angiogenesis therapy to induce early tumor response.Clinical trial registration Chinese clinical trial registry,ChiCRTONRC-12002394.

Objective:To investigate the effect of early debridement and open reduction combined with internal and external fixation on open fracture of tibia and fibula.Methods:The clinical data of 82 patients with open tibiofibular fractures admitted to the 901st Hospital of the Joint Logistic Support Force of the Chinese People′s Liberation Army from June 2017 to May 2018 were retrospectively analyzed. There were 42 males and 40 females, aged 20-62 years, with an average age (34.8±16.1) years. According to different surgical methods, they were divided into control group ( n=32) and observation group ( n=50). The patients in the control group received early debridement and limited internal fixation, the patients in the observation group received early debridement and open reduction combined with internal and external fixation. The operation time, blood loss, healing time, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) level, white blood cell count (WBC), excellent rate and complication rate were observed in both groups. The measurement data were expressed as mean±standard deviation( Mean± SD), comparison between groups was analyzed using independent sample t test, count data were expressed as percentage (%), comparison between groups was performed using chi-square test. Results:The operative time (98.35±15.14) min in the observation group were higher than those in the control group (79.26±13.22) min, blood loss (120.53±41.66) mL and healing time (16.84±5.07) min in the observation group were significantly lower than those in the control group [(79.26±13.22) min, (210.59±56.60) mL, (19.48±5.46) min]. The difference was statistically significant ( t values were 5.85, 7.76, 2.20, P<0.05). Compared with control group, the level of ESR (18.91±2.70) mm/h, CRP (39.20±3.13) ng/L, WBC (7.04±1.12)×10 3/L were significantly lower than control group [(27.36±3.28) mm/h, (45.63±4.06) ng/L, (11.06±1.51)×10 3/L]. The difference was statistically significant ( t values were 12.17, 7.63, 12.95, P<0.05). Compared with control groups′s excellent and good rate, the excellent and good rate of observation group was higher, but the difference was not statistically significant ( P>0.05). The incidence of complications in the observation group (6.00%, 3/50) was significantly lower than that in the control group (31.25%, 10/50) ( P<0.05). Conclusions:Early debridement and open reduction combined with internal and external fixation is an effective method for the treatment of open fracture of tibia and fibula. Compared with internal fixation, it has the advantages of shorter healing time, less blood loss and lower incidence of complications. And it can also reduce the inflammatory response of patients.

Objective:To investigate the clinical effect of free fibular flap of great toe without vein in repairing soft tissue defect of finger pulp.Methods:From June 2020 to January 2021, 8 cases of finger pulp soft tissue defect were repaired with free flap of fibular side of great toe without vein, including 6 males and 2 females, with an average age of 32(18-42) years old. The defect area of finger pulp was 1.0 cm×0.5 cm-1.8 cm×0.8 cm. The proper digital artery and digital nerve of the affected finger were anastomosed. All flaps did not carry reflux veins. The donor site of the toe was directly sutured. Postoperative routine anti infection, anti spasm and anticoagulant treatment was performed. Postoperative outpatient regular follow-up.Results:All flaps survived without vascular crisis, and the donor and recipient sites healed in the first stage. All patients were followed-up for an average of 6.4(4-12) months. At the last follow-up, the TPD was 6-8 mm, the hand function was evaluated according to the Trial Standard of Upper Limb Function Evaluation of Hand Surgery Society of Chinese Medical Association. Seven cases were excellent and 1 case was good.Conclusion:The free fibular flap of the great toe can be used to repair the small wound of finger pulp without carrying the draining vein, the flap can still survive. And it can provide a certain reference for the failure of anatomic separation of the draining vein.

Objective:To explore the relationship between expression levels of CLOCK mRNA and protein and the clinical characteristics of patients with nasopharyngeal carcinoma.Methods:The frozen tissue specimens from 33 patients with nasopharyngeal carcinoma in the Affiliated Tumor Hospital of Guizhou Medical University from 2018 to 2019 were collected. Seventeen cases of tissue specimens from patients with nasopharyngeal chronic inflammation in the Affiliated Hospital of Guizhou Medical University in 2019 were collected. From 2008 to 2014, 68 cases of formalin-fixed paraffin-embedding (FFPE) nasopharyngeal carcinoma tissue and 37 cases of FFPE nasopharyngeal chronic inflammation tissue were collected from the Affiliated Tumor Hospital of Guizhou Medical University. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot (WB) were used to detect the mRNA and protein expression levels of CLOCK. The nasopharyngeal carcinoma cells including CNE1, CNE2, 5-8F and the normal nasopharyngeal epithelial cell NP69 were cultured. qRT-PCR was used to detect the expression level of CLOCK mRNA in each cell line at the time points of ZT2, ZT6, ZT10, ZT14, ZT18 and ZT22. The cosine method was used to fit the rhythm of CLOCK gene in nasopharyngeal carcinoma. The protein expression of CLOCK protein was detected by using immunohistochemical method in 68 cases of nasopharyngeal carcinoma and 37 cases of nasopharyngeal chronic inflammation tissue. Survival was analyzed by Kaplan-Meier method and Log rank test, and the influencing factors was analyzed by Cox regression model.Results:The expression levels of CLOCK mRNA in CNE1, CNE2 and 5-8F cells (0.63±0.07, 0.91±0.02 and 0.33±0.04, respectively) were lower than that in NP69 cell (1.00±0.00, P<0.05). The expression levels of CLOCK protein in CNE1, CNE2 and 5-8F cells (0.79±0.06, 0.57±0.05 and 0.74±0.10, respectively) were lower than that of NP69 cells (1.00±0.00, P<0.05). The expressions of CLOCK mRNA in nasopharyngeal carcinoma cells including CEN1, CNE2, 5-8F and normal nasopharyngeal epithelial cell NP69 were different at different time points, with temporal fluctuations. The fluctuation periods of CLOCK mRNA in CNE1, CNE2, 5-8F, and NP69 cells were 16, 14, 22 and 24 hours, respectively. The peak and trough times were ZT10: 40 and ZT18: 40, ZT10 and ZT3, ZT14: 30 and ZT3: 30, ZT12: 39 and ZT0: 39, respectively. CLOCK mRNA and protein expression levels in nasopharyngeal carcinoma tissues (0.37±0.20 and 0.20±0.26, respectively) were lower than those in nasopharyngeal chronic inflammation tissues (1.00±0.00 and 0.51±0.41, respectively, P<0.05). The 1, 3, and 5-year survival rates of patients in the CLOCK protein high expression group (CLOCK protein expression level ≥ 0.178) were 96.2%, 92.1%, and 80.1%, respectively, which were higher than those in the low expression group (CLOCK protein expression level <0.178, 92.9% , 78.6% and 57.1%, respectively, P=0.009). The 1, 3, and 5-year progression-free survival (PFS) rates of patients in the CLOCK protein high expression group were 96.2%, 87.8%, and 87.7%, respectively, which were higher than those in the low expression group (92.7%, 82.2%, and 70.8%, respectively, P=0.105). Compared with the low-expression group (100.0%, 96.9%, and 90.0%, respectively), the 1, 3, and 5-year recurrence-free survival rates of patients in the CLOCK protein high expression group (100.0%, 95.7%, and 95.7%, respectively) were not statistically significant ( P=0.514). Compared with the low-expression group (92.7%, 82.2%, and 79.3%), the 1, 3, and 5-year survival rates without metastasis in the CLOCK protein high expression group (96.2%, 92.0%, and 92.0%, respectively) were not statistically significant ( P=0.136). CLOCK protein expression and T stage were independent prognostic factors of overall survival ( P<0.05). Conclusions:The expression of CLCOK is downregulated in the nasopharyngeal carcinoma cell and nasopharyngeal carcinoma tissues. Clock gene CLOCK is rhythmically expressed in the nasopharyngeal carcinoma cells and normal nasopharyngeal epithelial cells. Compared with normal nasopharyngeal epithelial cells, the fluctuation period of CLOCK in nasopharyngeal carcinoma cells is shortened. The overall survival of patients in the CLOCK protein high expression group is better than that of low expression group. The expression of CLOCK protein is an independent influencing factor for overall survival. CLOCK gene may be a potential tumor suppressor gene in the nasopharyngeal carcinoma.