Objective:To explore the spatial support capacity and its influence in osteogenic effect of composite biofilm based on poly(propylene carbonate)(PPC)/poly(butylene succinate)(PBS)in rabbit tibial bone defect models,and to clarify its barrier functional reliability and osteogenetic effect in vivo.Methods:The composite biofilms of PPC/PBS and PPC/PBS/collegen type Ⅰ(Col-Ⅰ)(PPC/PBS/Co)were prepared.Eighteen Japanese big-ear rabbits were selected and two bone defects were prepared on each side of the tibia of the rabbits.Six rabbits were randomly selected to place PPC/PBS composite biofilm on the bone defects,2 rabbits were executed at 2,4,8 and 12 weeks after operation,and the surface microstructures of PPC/PBS composite biofilm in the rabhit bone defect area were observed by scanning electron microscope(SEM).The experiment was divided into blank control group,PPC/PBS composite biofilm group,BME-10X collagen membrane group,and PPC/PBS/Co composite biofilm group.The above biofilms were placed on the corresponding bone defects of rabbits by operation,while no biofilm was placed in the rabbits in blank control group.Three rabbits were killed at 2,4,8 and 12 weeks after operation respectively,and the gray values of regenerated bone in the bone defect areas of the rabbits in varrous groups were detected by soft X-ray;the fluorescence intensities of regenerated bone tissue in the bone defect areas of the rabbits in various groups were observed by laser scanning confocal microscope after fluorescence labeling.The pathomorphology of regenerated bone tissue in the bone defect areas of the rabbits in various groups were observed by HE staining and modified Gomori staining,and the expression levels of bone morphogenetic protein 2(BMP-2)and osteopontin(OPN)in the regenerated bone tissue in bone defect areas of the rabbits in various groups were detected by immunohistochemical staining.Results:In general,the PPC/PBS composite biofilm was tightly covered in the bone defect area without displacement and collapse.The SEM results showed that the porous surface of PPC/PBS composite biofilm appeared micropore structure and the number of micropores was increased with the prolongation of time,while the smooth surface of biofilm basically did not form the micropore-like structure.The results of soft X-ray detection showed that the gray values of regenerated bone tissue in bone defect areas of the rabbits in various groups were increased with the prolongation of time,and the gray value of regenerated bone tissue in bone defect areas of the rabbits in PPC/PBS/Co composite biofilm group was significantly higher than those in other groups(P＜0.05).The confocal micrscope results showed that the fluorescence intensity of regenerated bone tissue in bone defect areas of the rabbits in PPC/PBS/Co composite biofilm group was similar to those in blank control group at 4,8,and 12 weeks;compared with PPC/PBS composite biofilm group and BME-10X collagen membrane group,the fluoresence intensity of regenerated bone tissue in bone defect areas of the rabbits in PPC/PB/Co composite biofilm group at 4 weeks was increased(P＜0.05),and the fluoresence intensity of regenerated bone tissue in bone defect areas of the rabbits at 8 and 12 weeks were decreased(P＜0.05).The results of HE staining and modified Gomori staining showed that compared with PPC/PBS composite biofilm group and BME-10X collagen membrane group,the new bone formed faster in PPC/PBS/Co composite biofilm group and blank control group at 2 and 4 weeks,and the lamellar bone mineralization was higher at 12 weeks.The immunohistochemical staining results showed that compared with blank control group,PPC/PBS composite biofilm group and BME-10X collagen membrane group,the expression levels of BMP-2 and OPN proteins in the regenerated bone tissue in bone defect areas of the rabbits in PPC/PBS/Co composite biofilm group at 2 and 4 weeks were increased(P＜0.05 or P＜0.01);compared with blank control group and PPC/PBS composite biofilm group,the expression levels of BMP-2 and OPN proteins in the regenerated bone tissue in bone defect areas of the rabbits in BME-10X collage membrane group were decreased(P＜0.05 or P＜0.01).Conclusion:PPC/PBS composite biofilm has excellent spatial support capacity and reliable physical barrier function.The PPC/PBS/Co composite biofilm has a good effect in guiding bone regeneration in vivo.

Cancer immunotherapy has become a promising strategy. However, the effectiveness of immunotherapy is restricted in "cold tumors" characterized with insufficient T cells intratumoral infiltration and failed T cells priming. Herein, an on-demand integrated nano-engager (JOT-Lip) was developed to convert cold tumors to hot via "increased DNA damage and dual immune checkpoint inhibition" strategy. JOT-Lip was engineered by co-loading oxaliplatin (Oxa) and JQ1 into liposomes with T-cell immunoglobulin mucin-3 antibodies (Tim-3 mAb) coupled on the liposomal surface by metalloproteinase-2 (MMP-2)-sensitive linker. JQ1 inhibited DNA repair to increase DNA damage and immunogenic cell death (ICD) of Oxa, thus promoting T cells intratumoral infiltration. In addition, JQ1 inhibited PD-1/PD-L1 pathway, achieving dual immune checkpoint inhibition combining with Tim-3 mAb, thus effectively promoting T cells priming. It is demonstrated that JOT-Lip not only increased DNA damage and promoted the release of damage-associated molecular patterns (DAMPs), but also enhanced T cells intratumoral infiltration and promoted T cell priming, which successfully converted cold tumors to hot and showed significant anti-tumor and anti-metastasis effects. Collectively, our study provides a rational design of an effective combination regimen and an ideal co-delivery system to convert cold tumors to hot, which holds great potential in clinical cancer chemoimmunotherapy.

Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases.The stability and sensitivity were evaluated using an accelerated thermostability test. Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. Conclusions The spbELISA has practical applications in assessing the vaccination status of large pig herds.

Objective:To establish the sex-, age-and season-specific (month) reference intervals (RI) for thyroid stimulating hormone (TSH) measurement by big data and indirect method in adults.Methods:TSH data of anonymous patients were collected from Beijing Chaoyang Hospital Affiliated to Capital Medical University in 2016, the data were selected and outliers were removed. Indirect methods (Hoffmann method and Bhattacharya method) were used to calculate TSH reference intervals of whole population, different genders, ages and seasons (months). TSH RI from two indirect methods of total population, selected population, physical examination population was compared with RI from reagent instruction according to reference change value ( RCV) based on biological variability. Results:A total of 61 599 records were obtained from 90 699 records including 18 776 males and 42 823 females. The TSH RI were obtained by Hoffmann method: the whole population, 0.59-5.59 μIU/ml (1 μIU/ml=1 mIU/L), male, 0.53-5.16 μIU/ml, female, 0.59-6.11 μIU/ml. The upper limits of TSH RI were higher with age and in winter (January): 18-30 years old, 0.62-5.57 μIU/ml, 71-80 years old, 0.49-6.45 μIU/ml; January, 0.59-6.40 μIU/ml, August, 0.60-5.56 μIU/ml; The RI of TSH by Bhattacharya method: the whole population, 0.58-5.80 μIU/ml, male, 0.55-5.02 μIU/ml, female, 0.62-6.21 μIU/ml. The upper limits of TSH RI were also higher with age and in winter (January): 18-30 years old, 0.65-5.67 μIU/ml, 71-80 years old, 0.46-5.99 μIU/ml, January: 0.61-6.52 μIU/ml, August: 0.61-5.69 μIU/ml. Compared to RI from reagent instruction, the differences of TSH RI from two indirect methods of total population, selected population, physical examination population were acceptable.Conclusions:TSH RI was established by indirect method. With the increase of age and winter, the upper limit of TSH reference interval tends to increase.

Objective:To investigate the application effect of bilobed skin flap with peroneal artery perforation in repairing postoperative defects of tongue cancer and oral floor cancer.Methods:Six patients with oral squamous cell carcinoma (4 males and 2 females, aged 28-72 years) admitted to the Department of Oral and Maxillofacial Surgery, the First Affiliated Hospital of Benbu Medical College from May 2017 to December 2019 were repaired with the peroneal artery perforator bilobed flap, one lobe repaired the tongue, and the other repaired the floor of the mouth. The donor area is directly closed and sutured. Postoperative follow-up was conducted to observe tumor recurrence and metastasis, and the patient’s appearance, mouth opening, swallowing function, speech function and lower limb recovery were evaluated.Results:All the 6 flaps survived. One of the patients had a single flap because the pedicle of the perforator vessel was not long enough. The other 5 cases were repaired with bilobed leaf flap. All the flaps survived and the wound surface healed in the first stage. One case was infected with effusion after operation, which was healed by washing and dressing change. The donor area can be sutured directly. No tumor recurrence or metastasis occurred 6 months after surgery. Speech, swallowing and lower limb movement basically returned to normal, facial morphology was basically symmetrical, and the degree of opening Ⅱ-Ⅲ. Six months after the operation, 5 patients with bilobed valve returned to a nearly normal state with a degree of opening Ⅱ-Ⅲ. One patient with single lobe recovered moderately, but speech and swallowing were still restricted. In 6 cases, the donor scar was concealed and the lower limb was basically normal.Conclusions:The bilobed flap with peroneal artery perforation can be used to repair postoperative defects of tongue carcinoma and floor of mouth carcinoma.

Objective:To investigate the application effect of bilobed skin flap with peroneal artery perforation in repairing postoperative defects of tongue cancer and oral floor cancer.Methods:Six patients with oral squamous cell carcinoma (4 males and 2 females, aged 28-72 years) admitted to the Department of Oral and Maxillofacial Surgery, the First Affiliated Hospital of Benbu Medical College from May 2017 to December 2019 were repaired with the peroneal artery perforator bilobed flap, one lobe repaired the tongue, and the other repaired the floor of the mouth. The donor area is directly closed and sutured. Postoperative follow-up was conducted to observe tumor recurrence and metastasis, and the patient’s appearance, mouth opening, swallowing function, speech function and lower limb recovery were evaluated.Results:All the 6 flaps survived. One of the patients had a single flap because the pedicle of the perforator vessel was not long enough. The other 5 cases were repaired with bilobed leaf flap. All the flaps survived and the wound surface healed in the first stage. One case was infected with effusion after operation, which was healed by washing and dressing change. The donor area can be sutured directly. No tumor recurrence or metastasis occurred 6 months after surgery. Speech, swallowing and lower limb movement basically returned to normal, facial morphology was basically symmetrical, and the degree of opening Ⅱ-Ⅲ. Six months after the operation, 5 patients with bilobed valve returned to a nearly normal state with a degree of opening Ⅱ-Ⅲ. One patient with single lobe recovered moderately, but speech and swallowing were still restricted. In 6 cases, the donor scar was concealed and the lower limb was basically normal.Conclusions:The bilobed flap with peroneal artery perforation can be used to repair postoperative defects of tongue carcinoma and floor of mouth carcinoma.

Objective To investigate the value of heparin-binding protein(HBP) in predicting urosepsis after endoscopic lithotripsy for upper urinary tract stones.Methods From January to November 2018,635 cases of upper urinary tract stones treated by endoscopic lithotripsy in our hospital was analyzed.According to the 2014 edition Chinese Guidelines of the diagnosis and treatment of urosepsis,all patients were divided into two groups,the definite urosepsis group (n =53,8.3％) and the no urosepsis group (n =582,91.7％).The serum of HBP level was detected by fluorescence immunoassay technology.The serum of procalcitonin (PCT) level was detected by cyclic enhanced immunofluorescent assay.C-reactive protein (CRP) level was detected by automatic chemiluminescence immunoassay analyzer.White blood cells (WBC) count was performed by hematology analyzer.Receiver operating characteristic(ROC) curves were constructed by SPSS 22 and MedCale,and areas under curves (AUC) were calculated and compared to illustrate the diagnostic power of HBP,PCT,CRP and WBC.The correlation between HBP and WBC was also significant (r =0.68,P ＜ 0.05).Results The level of HBP,PCT,CRP and WBC were significantly higher in the definite urosepsis group compared to the no urosepsis group.AUC of HBP,PCT,CRP and WBC were 0.930,0.878,0.727 and 0.617.When the cutoff of HBP,PCT,CRP and WBC were set as 51.5 ng/ml,0.41 ng/ml,64.2 mg/L and 11.7 × 109/L,the sensitivities were 89.2％,83.3％,83.5％ and 44.7％,and the specificity were 95.9％,75.6％,64.3％ and 77.1％,respectively.Conclusions The value of heparin-binding protein was superior to that of PCT,CRP and WBC in predicting urosepsis after endoscopic lithotripsy for upper urinary tract stones.Heparin-binding protein could be acted as a diagnostic marker for urosepsis.

To evaluate the clinical effect of application of free peroneal artery perforator flap in repairing postoperative tissue defect after resection of oral maxillary malignant tumor. Methods From January, 2016 to June, 2018, 14 patients with maxillary malignant tumor were treated with tumor excision. Then the free per-oneal artery perforator flap was used to reconstruct tissue defect caused by tumorectomy. There were 6 cases of squa-mous-cell carcinoma of palatine, 2 cases of adenoid cystic carcinoma of palatine, and 6 cases of maxillagingiva squa-mous cell carcinoma. The incised area of flap was 4.0 cm×5.0 cm to 7.0 cm×8.0 cm. There were 5 cases of hard palate soft tissue defect repair and 9 cases of simultaneous repair of soft and hard palate. Followed-up by outpatient service in 3-12 months after operation, postoperative maxillamorphology, phonetic function, swallow function, opening degree and prognosis of the patients were evaluated. Results All 14 implanted flaps were alive.One flap had vas-cular crisis, and rescued by surgical exploration and timely rescue.Three flaps were prolonged healing.All donor sites were sutured directly.All surgical incisions healed primarily.Half a year after the operation, the appearance of maxilla was formed gradually. The phonetic function, swallowing function, opening degree and movement of lower leg were all recovered normal. One year after the operation, epithelization was done in 6 cases. And there was no tumor recur-rence.Conclusion The peroneal artery perforator flap has long vascular pedicle, larg diameter, high survival rate after vascular anastomosis and relatively concealed donor area. It can be used to repair and reconstruct the tissue defect in maxillary malignant tumor resection and achieved good result.

In this paper, we developed an accurate and sensitive LC-MS/MS method for the determination of amoxicillin and clavulanic acid in human plasma. A 50% aqueous acetic acid solution was used as a stabilizer, and the plasma samples were evaporated to dryness and resolved after protein precipitation on ice bathing and then were placed in an autosampler for injection. The gradient was eluted by Hedera ODS-2 column(2. 1 mm×150 mm). The aqueous phase was an aqueous solution containing 0. 2% acetic acid. The organic phase was methanol. The amoxicillin and clavulanic acid were detected under negative ion detection with electrospray ionization(ESI)in multiple reaction monitoring(MRM)mode of m/z 364. 1→223. 1 and 198. 1→135. 9 in the triple quagdrupole tandem mass spectrometer(Triple Quad TM 6500+). The concentration ranges of plasma from 20. 0 ng/mL to 5 000 ng/mL for amoxicillin and 10. 0 ng/mL to 2 500 ng/mL for clavulanic acid were good linear relationship. The accuracy deviation were ±15. 0% and precision were less than 15. 0% for the intra-assay and inter-assay. The matrix effect and recovery meeted the acceptance criteria, amoxicillin and clavulanic acid were stable under storage and processing conditions. Healthy subjects were given a test preparation of amoxicillin and clavulanate potassium granules 1 bag(125 mg/31. 25 mg/bag)and the reference preparation amoxicillin clavulanate potassium dry mix Suspension “Augmentin® ” 5 mL(125 mg/31. 25 mg/5 mL)was used to determine the plasma concentration of amoxicillin and clavulanic acid. The Phoenix WinNonlin 6. 4 software was used to estimate the pharmacokinetic parameters of non-compartmental models. The pharmacokinetic parameters of amoxicillin and clavulanic acid were statistically calculated and evaluated the bioequivalence. what′s more, we evaluated the diet on the pharmacokinetics of amoxicillin and clavulanic acid. The analytical method was rapid and sensitive, which was successfully employed in the bioequivalence study of amoxicillin(125 mg/bag)and clavulanate potassium granules(31. 25 mg/bag)for determining the concentration of amoxicillin and clavulanic acid.

Objective To investigate the efficacy and safety of Nocardiarubra cell wall skeleton (N-CWS) bladder irrigation in prevention of recurrence after transurethral resection for the treatment of non-muscle invasive bladder cancer (NMIBC).Methods The clinical data of patients with NMIBC treated by N-CWS and epirubicin collected between October 2013 and November 2018 at the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed.All patients underwent TURBT.Among the 118 NMIBC patients,the average age was (65.1 ± 1 1.9) years,and the sex ratio (male/female) was 1.9∶1 (77/41).Patients were divided into two group:N-CWS group (n =55) and epirubicin group (n =63) according to different instillation regimens.N-CWS was given as an instillation of 800 μg in 50 ml of saline and maintained in the bladder for 2 h in the N-CWS group.Epirubicin was given as an instillation of 50 mg in 50 ml of saline and maintained in the bladder for 1 h in the epirubicin group.In the N-CWS group,mean agewas (64.9 ± 12.1) years and 37 (67.3％) were male.Multiple tumors were present in 17 (69.1％) patients.Tumor size was ≤3 cm in 49(89.1％) and 7(12.7％) had a history of NMIBC.Stage was Ta and T1 in 36(65.5％) and 19(34.5％),respectively.Grade 1,Grade 2 and Grade 3 were the primary grades in 38(69.1％),13(23.6％) and 4(7.3％),respectively.Low risk,intermediate risk and high risk were present in 14 patients(25.5％),16 (29.1％) and 25 (45.5％),respectively.In the epirubicin group,mean age was (65.3 ± 11.2) years and 40(63.5％)were male.Multiple tumors were present in 19(30.2％) patients.Tumor size was ≤3 cm in 56(88.9％) and 11 (17.5％) had a history of NMIBC.Stage was Ta and T1 in 37(58.7％) and 26 (41.3％),respectively.Grade 1,Grade 2 and Grade 3 were the primary grades in 44(69.8％),12(19.0％)and 7(11.1％),respectively.Low risk,intermediate risk and high risk were present in 13 (20.6％),19 (30.2％) and 31 (49.2％),respectively.The tumor recurrence,progression and adverse reactions after Intravesical Instillation in both groups were followed up and recorded.No significant differences were found between the two groups.Results A total of 118 patients were followed up.Mean follow-up time was (33.7 ± 5.4) months.25.5％ (14/55) in the N-CWS group vs.42.8％ (27/63) in the epirubicin group had recurrence after 5 years (x2 =3.922,P =0.048).The five-year RFS was higher in the N-CWS group than in the epirubicin group (74.2％ vs.56.5％,P =0.044).No significant difference was found in the progression rate between the two groups(5.5％ vs.7.9％,P =0.867).The incidences of adverse events in the two groups were 16.4％ (9/55) and 19.0％ (12/63),respectively.The N-CWS group had significantly fewer cases with urinary frequency and dysuria than the epirubicin group.No significant differences were found in other side effects.Conclusions Intravesical instillation of N-CWS after NMIBC TURBT was found to be a promising procedure to prevent recurrence and prolong the recurrence-free survival with less side effects.