ObjectiveTo investigate the effects of thyme herbal tea （BLX） on the proliferation of human coronavirus OC43 （HCoV-OC43） in vitro and in vivo. MethodThe chemical composition of BLX was analyzed by UPLC-MS. The cytotoxicity of BLX in HRT-18 cells and the effect of BLX treatment on the proliferation of HCoV-OC43 in cells were analyzed. Copies of viral gene were detected by real-time PCR. The effect of BLX treatment on the life cycle of HCoV-OC43 was detected by time-of-addition assay. The maximum tolerated dose of BLX and the influences of BLX on the body weight and survival time of suckling mice infected with HCoV-OC43 were determined. The expression of viral protein in the brain and lung tissue was analyzed by immunohistochemistry. ResultThere were 11 chemical components identified in BLX by UPLC-MS. BLX showed the 50% cytotoxic concentration （CC₅₀） of （13 859.56±319） mg·L^-1， the median inhibitory concentration （IC₅₀） of （1 439.09±200） mg·L^-1， and the selection index of 8.26-11.44 for HCoV-OC43 in HRT-18 cells. Compared with the cells infected with HCoV-OC43， BLX at the concentrations of 1 500， 1 000， 500 mg·L^-1 inhibited the proliferation of this virus （P<0.05， P<0.01）. BLX exhibited antiviral effect in the early stage of virus infection， and the inhibition role in the attachment stage was more significant than that in the entry stage （P<0.05）. In the suckling mice infected with HCoV-OC43， BLX at 1200 and 600 mg·kg^-1·d^-1 alleviated the symptoms， prolonged the survival period， reduced the death rate， and down-regulated the mRNA level of nucleocapsid protein in the mice. Moreover， BLX at 1 200 mg·kg^-1·d^-1 down-regulated the expression of nucleocapsid protein in the brain （P<0.01） and the lung （P<0.01）. ConclusionBLX contained multiple antiviral ingredients. It inhibited the proliferation of HCoV-OC43 both in vitro and in vivo by interference with viral attachment. This study provides theoretical reference for the treatment of acute respiratory tract infection with HCoV-OC43 and for further development and application of BLX.

Background: and Purpose Brainstem gliomas (BSGs) in adults are rare brain tumors with dismal outcomes. The aim of this study was to determine the clinical and genetic features in a series of BSGs and their association with the prognosis. Methods: Fifty patients who underwent a stereotactic biopsy between January 2016 and April 2018 at a single institution were collected. Data on clinicopathological characteristics were analyzed and factors associated with patient survival were identified using a Cox regression model. Results: The median age at diagnosis was 55.5 years, and 62% of the patients were male. Glioblastoma (44%) accounted for the largest proportion of BSGs, and oligodendroglioma (2 of 50) was rarely encountered. The IDH mutation (6 of 44) occurred infrequently in astrocytomas, and IDH-mutant tumors harbored both ATRX loss and MGMT promoter methylation at a relatively low level. Wild-type IDH astrocytomas were identified as having high rates of 1p/19q codeletion (5 of 38) and loss of heterozygosity 1p (8 of 38) or 19q (8 of 38) only. In diffuse midline glioma H3K27M mutant, MGMT promoter methylation occurred in three of four cases. Patients were offered radiotherapy and/or concurrent/adjuvant temozolomide chemotherapy, and their median survival time was 13 months. Multivariate analysis revealed that a low tumor grade, absence of tumor enhancement, duration of symptoms ≥3 months, Karnofsky performance status ≥70, and ATRX loss conferred a survival advantage. Conclusions Adult BSGs showed different molecular genetic characteristics, but also resembled supratentorial gliomas in their clinical features associated with oncological outcomes.

Background: and Purpose Brainstem gliomas (BSGs) in adults are rare brain tumors with dismal outcomes. The aim of this study was to determine the clinical and genetic features in a series of BSGs and their association with the prognosis. Methods: Fifty patients who underwent a stereotactic biopsy between January 2016 and April 2018 at a single institution were collected. Data on clinicopathological characteristics were analyzed and factors associated with patient survival were identified using a Cox regression model. Results: The median age at diagnosis was 55.5 years, and 62% of the patients were male. Glioblastoma (44%) accounted for the largest proportion of BSGs, and oligodendroglioma (2 of 50) was rarely encountered. The IDH mutation (6 of 44) occurred infrequently in astrocytomas, and IDH-mutant tumors harbored both ATRX loss and MGMT promoter methylation at a relatively low level. Wild-type IDH astrocytomas were identified as having high rates of 1p/19q codeletion (5 of 38) and loss of heterozygosity 1p (8 of 38) or 19q (8 of 38) only. In diffuse midline glioma H3K27M mutant, MGMT promoter methylation occurred in three of four cases. Patients were offered radiotherapy and/or concurrent/adjuvant temozolomide chemotherapy, and their median survival time was 13 months. Multivariate analysis revealed that a low tumor grade, absence of tumor enhancement, duration of symptoms ≥3 months, Karnofsky performance status ≥70, and ATRX loss conferred a survival advantage. Conclusions Adult BSGs showed different molecular genetic characteristics, but also resembled supratentorial gliomas in their clinical features associated with oncological outcomes.

Objective:To study the effects of enoxaparin on osteogenic differentiation of bone marrow stem cells (BMSCs) and the exosomes derived from BMSCs.Methods:After the BMSCs from 4-week old male SD rats were cultured, their surface antigen and multilineage differentiation potentials were identified. Subsequently, the BMSCs were incubated with osteogenic differentiation medium containing 10 IU/mL enoxaparin for 14 days. After the exosomes derived from BMSCs (BMSC-Exos) were extracted by the kit method, their structure was observed by transmission electron microscopy and their surface antigen CD63 detected by Western blot. Alizarin red staining was used to analyze the osteogenic differentiation of BMSCs. Osteogenic proteins including OCN and BMP-2 in BMSCs and exosomes were detected.Results:The spindle-shaped BMSCs isolated and cultured showed a uniform spiral pattern. They expressed highly the surface markers CD29 and CD44 but lowly CD34 and CD45, indicating that the majority of the cells were BMSCs. BMSC-Exos, in an oval shape with a diameter of about 30 to 80 nm, expressed CD63. Alizarin red staining showed that the number of mineralized nodules in the enoxaparin treatment group was significantly lower than that in the control group. Western blot analysis indicated that enoxaparin inhibited the expression of osteocalcin (OCN) and BMP-2 in BMSCs. ELISA results showed that the protein levels of OCN(48.81 ng/mL ± 8.23 ng/mL) and BMP-2 (311.45 pg/mL ± 27.59 pg/mL) in the BMSCs treated with enoxaparin were significantly lower than those of OCN (80.43 ng/mL ± 10.74 ng/mL) and BMP-2 (399.23 pg/mL ± 32.25 pg/mL) in the control BMSCs ( P<0.05). The contents of OCN (1.45 ng/mL±0.15 ng/mL) and BMP-2 (18.47 pg/mL ± 0.54 pg/mL) in the exosomes from BMSCs treated with enoxaparin were significantly higher than those of OCN (1.00 ng/mL ± 0.12 ng/mL) and BMP-2 (9.07 ng/mL ± 0.36 pg/mL) in the exosomes from control BMSCs ( P<0.05). Conclusions:Enoxaparin may inhibit the osteogenic differentiation of BMSCs. The mechanism of this might be related to its direct inhibition against the expression of OCN and BMP-2 in BMSCs and its indirect reduction of OCN and BMP-2 in BMSCs through the exhaust of the above proteins in the form of exosomes.

Objective To evaluate the clinical and radiologic outcomes of minimally invasive plate percutaneous osteosynthesis (MIPPO) for distal clavicle fractures of Neer type Ⅱ. Methods From May 2015 to May 2016, 13 patients with distal clavicle fracture of Neer type Ⅱwere treated with MIPPO using an anatomic locking plate. Anteroposterior plain X-ray images of the clavicle were taken at intervals until union was observed. Fracture healing time and complications were recorded, and shoulder joint functions evaluated using Constant and University of California at Los Angeles (UCLA) scores. Results The average operating time was 60.9 minutes (range, from 48 to 83 minutes), average intraoperative blood loss 24.7 mL (range, from 18 to 30 mL), and average union time for the patients was 13.3 weeks (range, from 11 to 15 weeks). No delayed union or nonunion occurred. There were no major complications, like infection, plate breakage, or neurovascular injury. Only 2 patients felt local incision numbness. All the patients were satisfactory with their shoulder func-tions. At the final follow-ups, the mean Constant score was 93.2 points (from 87 to 95 points) and the mean UCLA score 30.6 points (from 28 to 34 points). Conclusion In the treatment of distal clavicle fractures of Neer type Ⅱ, minimally invasive plate percutaneous osteosynthesis can not only achieve good clinical and radi-ologic outcomes but also be advantageous over conventional open reduction in aspects of smaller invasion and better cosmetic effect.

Objective To establish an enterovirus 71(EV71) infection model of tree shrew primary renal cells.Methods Tree shrew primary renal cells were obtained by trypsin digestion.After subculture and purification,EV71 virus was used to infect these primary cells.The culture supernatant of these EV71-infected cells was collected for virus titer detection at 1,2,4,6 and 8 days post-infection.The cells were collected for detection of EV71 VP1 protein by Western blot assay.Furthermore,the expression and location of VP1 protein in the infected cells were detected by indirect immunofluorescence assay.Vero cells were taken as positive control to evaluate the infectivity of EV71 virus to tree shrew primary renal cells.Results Morphologically,the cultured cells were proved to be majorly consisted of the primary renal cells after subculture and purification.The obtained primary cells were infected by EV71 virus.The virus titer was up to 1.3×106 TCID 50/mL during 48-96 h post-infection,proving that EV71 virus infected and proliferated in the tree shrew primary renal cells.Western blot showed that the viral VP1 protein was detected from infected primary cells at 2 to 8 d post infection.VP1 protein was also observed in the cytoplasm at 2 to 6 d post infection by indirect immunofluorescence.Compared with Vero cells,the infectivity of EV71 virus to tree shrew primary renal cells and its proliferation were confirmed.Conclusions Based on the successful establishment of cell culture of tree shrew primary renal cells,the infectivity to the obtained cells and proliferation of EV71 virus in the cells are confirmed.The model of EV71 virus-infected tree shrew primary renal cells is initially established.

Objective To compare the clinical outcomes of bone transport technique versus induced membrane technique for treatment of infected bone nonunion.Methods A retrospective study was made on 46 cases of infected bone nonunion admitted from April 2011 to December 2014.They were 33 males and 13 females,aged from 12 to 68 years (mean,45.3 years),involving 40 tibial and 6 femoral fractures.Bone transport technique was used in 37 cases while 9 cases were treated with induced membrane technique.Fracture union and complications were recorded.The function of the affected limb was evaluated at the final follow-up using Johner-Wruhs criteria.Results The 46 cases were followed up for 9 to 18 months (mean,14 months).In the bone transport group,bone union was achieved after a mean duration of 6.5 months (range,from 4 to 11 months) without complications like wound infection or neurovascular lesions.In the induced membrane group,bone union was achieved after a mean duration of 5.8 months (range,from 4 to 11 months) and there was one case of re-infection.According to the Johner-Wruhs score system,34 cases were rated as excellent,9 as good and 3 as fair,with a total excellent and good rate of 93.5％.Bone transport technique led to 28 excellent,4 good and one fair cases in the patients with tibial nonunion,and one excellent,2 good and one fair cases in the patients with femoral nonunion.Induced membrane technique led to 3 excellent,3 good and one fair cases in the patients with tibial nonunion,and 2 excellent cases in the patients with femoral nonunion.Conclusions Both bone transport technique and induced membrane technique are effective for infected bone nonunion.Bone transport technique may be preferred for tibial infected nonunion while induced membrane technique preferred for femoral infected nonunion.

Objective To explore the clinical efficacy of locking plate in the treatment of postoperative nonunion of limb long bone fracture.Methods We treated 53 patients whose limb long bone fracture failed to unite after operation with locking plate between April 2011 and November 2014.They were 38 men and 15 women,aged from 20 to 69 (average,39.7).There were 18 humeral fractures,5 radioulnar ones,9 femoral ones,and 21 tibial ones.On average,each patient had undergone 1.2 previous operations (from 1 to 2 times),including plating in 29 cases,intramedullary nailing in 15,and external fixation in 9.In the 15 patients whose intramedullary nails were reserved because of slightly loosening only under axial stress,lateral force and rotation,the nonunion was treated with bone autograft + unicortical or full-thickness locking plate.In the other 38 patients whose original implants were removed,the nonunion was treated by re-reaming of the medullary cavity + bone autograft + locking plate internal fixation.Regular radiological follow-ups were conducted for the patients.The functional recovery of upper limbs was evaluated by the Neer system while that of the lower limbs by the Johner-Wruhs system.Results One patient was lost to the follow-up.The rest 52 were followed up for 4 to 15 months (average,12.7 months).Nonunion was found in 2 patients whose internal fixation was broken and loosened.Bone healing was achieved in 50 cases after 4 to 6 months(average,5 months).Removal of implants was conducted in 36 patients at 12 to 24 months after operation (average,13.3 months).At the final follow-ups,the Neer score of upper limb was excellent in 15 cases,good in 5,fair in one and poor in one,giving an excellent to good rate of 90.9％;the Johner-Wruhs score of lower limb was excellent in 24 cases,good in 3,fair in 2 and poor in one,giving an excellent to good rate of 90.0％.Conclusion Locking plate can have good curative efficacy in the treatment of postoperative nonunion of long bone fracture.

BACKGROUND:The in vivo degradation process of chitosan/hydroxyapatite composite porous scaffolds is not very clear. Research on the effects of rat osteoblasts and degradation products is less.
OBJECTIVE:To analyze the biocompatiblity of rat osteoblasts with degradation products of chitosan/hydroxyapatite composite porous scaffolds.
METHODS:The second generation of cultured rat osteoblasts were respectively cultured in the extract of degradation products of chitosan/hydroxyapatite composite scaffolds (experimental group) and Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum (control group). At 2, 4, 6, 8, 10 days of culture, cel counting was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Alkaline phosphatase activity was measured by the recommended method of determination of the Federation, and total protein was determined by BCA method.
RESULTS AND CONCLUSION:The proliferation speed, alkaline phosphatase activity, total cel ular protein synthesis and ratio of alkaline phosphatase to total protein in rat osteoblasts cultured in the experimental group were significantly higher than those in the control group (P<0.05). This experiment showed that the degradation products of chitosan/hydroxyapatite composite porous scaffolds cannot only promote rat osteoblast adhesion, growth and proliferation, but also enhance its ossification function, with good biocompatibility.

BACKGROUND:Nowadays, the internal fixation materials for the clinical treatment of unstable femoral intertrochanteric fractures are broadly divided into two categories:intramedul ary fixation system and
extramedul ary fixation system. However, the effects of the treatments usual y lack of macro evaluation.
OBJECTIVE:To compare the effect of intramedul ary fixation system and extramedul ary internal fixation system in the treatment of unstable femoral intertrochanteric fractures.
METHODS:217 patients with unstable femoral intertrochanteric fracture (Evans typing: Ⅲ type to Ⅴ type and R type) were treated by using intramedul ary fixation system and extramedul ary internal fixation system:18
cases were treated with dynamic condylar screw, 67 cases were treated with dynamic hip screw, 43 cases were treated with reconstruction nail, 72 cases were treated with proximal femoral anti-rotation blade nail, and 17 cases were treated with new generation of antegrade interlocking intramedul ary nail. The operative time, length of
incision, blood loss, time in bed, fracture healing time, complications and Harris hip score were analyzed and compared.
RESULTS AND CONCLUSION:In the aspects of operative time, blood loss, time in bed, complications and
Harris hip score, the intramedul ary fixation system was superior to extramedul ary internal fixation system. But there were significant differences in the aspects of length of incision and fracture healing time between two
methods. The results showed that the treatment of unstable femoral intertrochanteric fractures by using
intramedul ary fixation system was better than the extramedul ary internal fixation system, as the intramedul ary fixation system could effectively shorten the operative time and reduce blood loss. This stable and reliable fixation enables patients to take early postoperative functional exercise, and could significantly shorten the time in bed and reduce the incidence of complications and it is conducive to postoperative hip function recovery.
Therefore, intramedul ary fixation system is an ideal method for the treatment of unstable femoral intertrochanteric fractures.