Transalveolar technique for sinus floor elevation(TSFE)offers the advantages of minimal invasiveness,reduced postoperative reaction,and shorter operative time for vertical bone augmentation in the maxillary posterior region.The clinical data of one patient with severe deficiency of residual bone height(RBH)in the maxillary posterior region,a blood vessel visible in the lateral wall of the maxillary sinus and a visible septum at the floor of the maxillary sinus were reported,and two-stage flexible TSFE was used to improve the vertical bone height of the operated area while reducing trauma,the risk of Schneiderian membrane rupture and maxillary sinus infection,etc.,and the relevant literatures were reviewed.The patient,male,26 years old,complained of missing left maxillary posterior teeth for more than 1 year and requested restoration.The patient had 27 missing teeth,normal keratinized gingiva,full alveolar ridge,no elongation of the opposing teeth,fair width of the proximal and normal occlusal distance.The results of cone beam CT(CBCT)showed that the distance between the sinus crests at the site of the 27 teeth was about 3 mm,the width of the alveolar bone was about 12.8 mm,the bone density was normal,and there were no residual roots or other abnormalities;no cyst-like lesions were seen in the walls of the maxillary sinuses bilaterally,and separation was seen at the floor of the maxillary sinus on the left side and a blood vessel was seen in the lateral wall of the maxillary sinus.A diagnosis of Kennedy class Ⅱ maxillary tooth defects was made.After two stages of TSFE,the Schneiderian membrane was intact and the bone height of the implant area was elevated to 9.6 mm from 3 mm preoperatively after the completion of the restoration,with stable bone augmentation,good osseointegration,and restoration of normal occlusal function.For the patients with severe bone height deficiency in the maxillary posterior region,flexible two-stage TSFE should be considered,which can help to reduce the risk of maxillary sinus infection and Schneiderian membrane rupture while minimizing the damage and obtaining the ideal bone augmentation results.

Objective To explore the molecular mechanism by which TP53 regulating pyroptosis,invasion and migration of embryonic fibroblasts through MMP1/NLRP3 signaling pathway.Methods NIH-3T3 murine embryonic fibroblasts were transfected with lentivirus and grouped as Control,Vector,oeTP53,oeTP53+shNC,and oeTP53+shMMP1.Cell proliferation and viability were assessed via CCK-8 assay,apoptosis by flow cytometry,and migration/invasion through scratch and invasion experiments.Protein interaction between P53 and MMP1 was confirmed by Co-immunoprecipitation.RT-qPCR evaluated mRNA expression of TP53,Collagen Ⅰ,Collagen Ⅲ,and α-SMA,while Western blot analyzed protein levels of these markers and pyroptosis-related proteins.Transmission electron microscopy was employed to examine cellular pyroptotic body modifications.Results Com-pared with Control and Vector groups,the oeTP53 group showed reduced cell proliferation activity(P<0.01),in-creased cell apoptosis rate(P<0.0001),decreased invasion(P<0.0001)and migration capabilities(P<0.0001);reduced Collagen I(P<0.001),Collagen Ⅲ(P<0.01),and α-SMA(P<0.01)protein expressions;increased NLRP3(P<0.05)and cleaved-caspase-1 expressions(P<0.01);and numerous pyroptotic bodies.MMP1 protein levels were found to be elevated in the oeTP53 group(P<0.05),and Co-IP demonstrated an interaction between p53 and MMP1 proteins.Compared with oeTP53 group,the oeTP53+shMMP1 group showed increased cell viability(P<0.001),decreased cell apoptosis rate(P<0.01),and increased cell migration(P<0.01)and invasion capabilities(P<0.01),increased scar formation-related protein expressions of Collagen I(P<0.01),Collagen III(P<0.001),and α-SMA(P<0.05);decreased pyroptosis-related protein expressions of NLRP3(P<0.01)and cleaved-caspase-1(P<0.001);and reduced pyroptotic bodies.Conclusion Overexpression of TP53 can inhibit mouse embryonic fibroblast proliferation,migration,and invasion,reduce scar formation-related protein expressions,and promote cell pyroptosis,with its mechanism potentially related to the MMP1/NLRP3 pathway.

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.

Cytopharmaceutical based on macrophages is a breakthrough in the field of targeted drug delivery. However, it remains a challenge to localize and control drug release while retaining macrophage activity and exerting its immunotherapeutic effect. Herein, a localized light-triggered release macrophage cytopharmaceutical (USIP@M) was proposed, which could utilize the tumor targeting and immunotherapy effects of macrophages to reverse the immune suppression of tumor microenvironment (TME). Amphiphilic block copolymers with ultraviolet (UV)-responsive o-nitrobenzyl groups were synthesized and co-loaded with sorafenib (SF), IMD-0354 (IMD), and upconverting nanoparticles (UCNPs), which were then taken up by macrophages, and the targeted delivery of drugs was realized by using the tumor tropism of macrophages. UCNPs converted near-infrared light with strong penetrability and high safety into UV light, which promoted the photoresponsive depolymerization of block copolymers and production of exosomes from USIP@M, accelerated drug efflux and maintained the activity of macrophages. IMD simultaneously polarized carrier macrophages and tumor-associated macrophages to exert the antitumor effect of macrophages, enhance T cell immunity, and alleviate the immunosuppressive state of TME. Synergistically with the chemotherapeutic effect of SF, it could effectively kill tumors. In conclusion, based on the localized light-triggered release strategy, this study constructed a novel macrophage cytopharmaceutical that could localize and control drug release while retaining the activity of macrophages and exerting its immunotherapeutic effect, which could effectively treat solid tumors.

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.

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.