Mutations, Recombination and Insertion in the Evolution of 2019-nCoV

BackgroundThe 2019 novel coronavirus (2019-nCoV or SARS-CoV-2) has spread more rapidly than any other betacoronavirus including SARS-CoV and MERS-CoV. However, the mechanisms responsible for infection and molecular evolution of this virus remained unclear. MethodsWe collected and analyzed 120 genomic sequences of 2019-nCoV including 11 novel genomes from patients in China. Through comprehensive analysis of the available genome sequences of 2019-nCoV strains, we have tracked multiple inheritable SNPs and determined the evolution of 2019-nCoV relative to other coronaviruses. ResultsSystematic analysis of 120 genomic sequences of 2019-nCoV revealed co-circulation of two genetic subgroups with distinct SNPs markers, which can be used to trace the 2019-nCoV spreading pathways to different regions and countries. Although 2019-nCoV, human and bat SARS-CoV share high homologous in overall genome structures, they evolved into two distinct groups with different receptor entry specificities through potential recombination in the receptor binding regions. In addition, 2019-nCoV has a unique four amino acid insertion between S1 and S2 domains of the spike protein, which created a potential furin or TMPRSS2 cleavage site. ConclusionsOur studies provided comprehensive insights into the evolution and spread of the 2019-nCoV. Our results provided evidence suggesting that 2019-nCoV may increase its infectivity through the receptor binding domain recombination and a cleavage site insertion. One Sentence SummaryNovel 2019-nCoV sequences revealed the evolution and specificity of betacoronavirus with possible mechanisms of enhanced infectivity.

Silicate-Cu/Mg bioactive ceramics promote the osteogenesis of osteoblasts / 中国组织工程研究

BACKGROUND: The use of silicate bioceramics as a tissue-engineered bone scaffold has poor ability to promote osteogenesis. Studies have shown that copper, magnesium, and other essential trace elements have obvious effects on the induction and stimulation of osteoblasts and hemangioblasts. OBJECTIVE: To investigate the effects of silicate bioactive ceramics with Cu and Mg on osteoblast proliferation and osteogenesis. METHODS: Cu-silicate bioceramics, Mg-silicate bioceramics, and Cu-Mg-silicate bioactive ceramics were prepared by the sol-gel method (molar ratio of both Cu and Mg in ceramics was 5%). Three experimental groups were CS-5Cu, CS-5Mg, CS-5Cu/5Mg groups. The silicate bioactive ceramics served as the control group (denoted as CS). X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy were used to characterize the samples. The surface crystallization of bioceramics was detected. Osteoblasts were co-cultured with four groups of ceramics for 24 hours. Osteoblast proliferation index, alkaline phosphatase secretion, osteopontin and osteocalcin gene expression, vinculin and actin protein expression were determined. RESULTS AND CONCLUSION: (1) The crystallization ability of different silicate bioceramic samples followed the order of CS-5Cu>CS>CS-5Cu/5Mg>CS-5Mg. (2) Osteoblast proliferation index followed the rule of CS-5Cu/5Mg>CS-5Cu≈CS-5Mg>CS. (3) Alkaline phosphatase secretion was in the order of CS-5Cu/5Mg>CS-5Cu≈CS-5Mg>CS. (4) Osteopontin and osteocalcin gene expression followed the rule of CS-5Cu/5Mg>CS-5Cu≈CS-5Mg>CS. (5) Vinculin and actin protein expression was in the order of CS-5Cu/5Mg>CS-5Cu≈CS-5Mg>CS. (6) These results suggest that Cu- or Mg-silicate, in particular Cu-Mg-silicate bioactive ceramics can promote the proliferation of osteoblasts and the expression of osteogenesis-related genes as well as cell adhesion and spreading.

Effects of computer aided design and manufacture for reconstruction of mandibular defect with vascu-laried iliac crest flap graft / 中华医学美学美容杂志

Objective To evaluate the clinical application and effect of computerized digital sur -gical assisted design and manufacture for reconstruction of mandibular defect with vascularied iliac crest flap graft .Methods CT was used to scan the craniofacial and maxillofacial region of mandibular ameloblastoma patient ,and then the 3D model of the maxillofacial skeleton was reconstructed using the computerized digital surgical aid design to simulate the focal resection and the mandibular defect reconstruction with the iliac crest flap .Surgical guides were prepared by 3D printing to assist focal re-section and bone reshaping .Postoperatively ,clinical and CT examinations were performed to observe the restoration of the patient's facial appearance and occlusal relationship ,as well as wound healing and graft survival .Results Postoperative review results showed that the bone graft survived well ,and the anastomosed bone was formed .The wound in the surgery area healed .No recurrence of the tumor was found after clinical and CT examination .The maxillofacial appearance was well restored ,and the re-sidual occlusal relationship recovered well .Conclusions The use of computerized digital surgical aided design combined with 3D printing technology can effectively simplify the mandibular segmental defect repair and reconstruction and optimize the postoperative repair effect .

Values of CAD/CAM technology and assistant reconstruction in mandible defect with fibular musculocutaneous flap / 中华医学美学美容杂志

Objective To explore the role of computer-aided design and manufacturing technology (CAD/CAM technology) in the repair of mandibular defects using the fibular osteomyocutaneous free flap.Methods Eight patients with a portion of mandible defect due to various reasons were subjected to preoperative CT scan,obtaining CT data of the mandible and the fibular bone to be transplanted.The original and the reconstructed skull model,as well as the mandible and fibula osteotomy guide plates,was printed using CAD/CAM technology.The titanium plates were preformed using the reconstructed skull model,and the operation was performed with the help of guide plates and a preformed titanium plate.The patients were followed up for 6 to 24 months to evaluate the operation effect.Results Two in 8 cases had flap necrosis caused by immune rejection of the implant,and one case had flap necrosis due to poor postoperative blood glucose control.The other 5 patients healed at the primary site and the fibular myocutaneous flap survived well.There were no serious complications in the donor and recepient site.The patients' satisfaction was high.Conclusions The reconstruction of mandible with the aid of CAD/CAM technique has the advantages of accelerating the operation process,shortening the operation time,improving the accuracy of mandibular amputation and fibular shaping,promotlng postoperative function and morphology recovery.

[Application of modified coronal approach in treatment of craniomaxillofacial fractures].

OBJECTIVE: This study aimed to investigate clinical efficacy of a modified coronal approach in treatment of craniomaxillofacial fractures. METHODS: Thirty-seven cases of craniomaxillofacial fractures underwent open reduction and rigid internal fixation through modified coronal approach. Clinical follow-up visits were conducted to evaluate facial nerve functional status, temporal shape, appearance restoration, and reduction of fracture and surgical area scars. RESULTS: During follow-up period of 6-36 months, appearance and function of all 37 patients recovered well without facial nerve injury and temporal depression deformity. All cases presented hidden scars, except for one case with hypertrophic scar. CONCLUSIONS: Applying modified coronal approach to craniomaxillofacial surgery effectively reduces incidence of temporal depression and facial nerve injury compared with traditional approach. The modified coronal approach produced more subtle scars compared with traditional approach and should be applied to treatment of craniomaxillofacial fractures.

Application of individualized anterolateral thigh flap (ALTF) designed by computed tomography angiography (CTA) for tongue reconstruction after resection of tongue carcinoma / 实用口腔医学杂志

Objective:To evaluate the clinical applicability of CT angiography (CTA) for locating the perforator vessels of anterolateral thigh flap(ALTF) and the effects of individualized ALTF designed by CTA for the reconstruction of the soft tissue defects of tongue after the resection of tongue carcinoma.Methods:21 patients with tongue squamous cell carcinoma underwent CTA for locating the perforator vessels of ALTFs and for the design of individualized ALTFs before operation.The patients underwent soft tissue defect reconstruction with individualized ALTFs after tumor removal.Results:All the operations came off as preoperative designed,the intraoperative findings of the blood vessel alignment were consistent with the preoperative CTA results.The size of flaps was 6.0 cm × 5.0 cm-11.0 cm× 8.5 cm.20 ALTFs survived,2 ALTFs appeared vascular crisis,1 remained survival and the other was necrotic after surgical exploration.During 6 ～ 60 momhs of follow-up,the survival condition of flaps and the wound healing condition were both satisfactory.1 patient died because of distant metastasis 18 months after operation.20 patients reminded free of carcinoma and satisfied with the reconstructive effects of chewing,swallowing and linguistic function.Conclusion:CTA can accurately locate perforator vessels for the design of individualized AFLT.AFLT designed by CTA is an ideal choice for the reconstruction of postoperative soft tissue defects after resection tongue carcinoma.

Loop-mediated isothermal amplification technique in the diagnosis of hepatitis B virus infection: a Meta-analysis / 中华预防医学杂志

Objective@#To evaluate the diagnostic performance of Loop-mediated isothermal amplification (LAMP) in the diagnosis of Hepatitis B Virus (HBV) infection using Meta-analysis.@*Methods@#Literatures about LAMP in the diagnosis of HBV throughPubMed database of the National Library of Medicine, the EMBASE database of the Dutch Medical Digest, the Cochrane Clinical Trials Database, China Science Periodical Database, CSPD and the China National Knowledge Infrastructure (CNKI) were searched from 2000 to 2016, and the Language limited to Chinese and English. English search terms include: LAMP, Loop-mediated isothermal amplification, HBV, hepatitis B virus; Chinese search terms include: loop-mediated isothermal amplification technology, HBV, hepatitis B virus. The keywords and free words are combined to search the literature, and the references mentioned in the retrieval literature are searched twice. The pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio (DOR), Q index as well as area under summary receiver operating characteristic curve (SROC) were calculated with Stata 12.0 software.@*Results@#A total of 12 literatures with 1 494 cases were included. The pooled sensitivity, specificity, positive likelihood ratio and negative likelihood ratio were 0.922 (95%CI: 0.905-0.937), 0.860 (95%CI: 0.818-0.896), 0.093 (95%CI: 0.048-0.182), and 15.400 (95%CI: 2.003-118.380), respectively. The DOR, area under SROC and Q index were 311.090 (95%CI: 95.841-1 009.800), 0.986 (95%CI: 0.974-0.998) and 0.949 (95%CI: 0.922-0.976), respectively. Deek's test indicates that no publication bias were found (P=0.140).@*Conclusion@#LAMP is worth to be popularized in field tests and primary-level hospitals tests.

Application of an individualized titanium mesh based on digital model in the repair of orbital fracture / 中国组织工程研究

BACKGROUND: Three-dimensional printing to prepare a digital model can improve the accuracy of orbital fracture repair, and has a good clinical prospect.OBJECTIVE: To evaluate the effect of individualized titanium mesh based on the digital model in the reconstruction of orbital fracture.METHODS: Twelve cases of orbital fracture were admitted at the Hospital of Stomatology, Southwest Medical University from January 2014 to October 2015. CT scanning in axial, coronal, sagittal planes and three-dimensional reconstruction were performed routinely in all cases preoperatively. Digital model was designed by 3D printing technology according to the CT data. Individualized titanium mesh was shaped based on the digital model and used to repair orbital fracture. The accuracy of the reconstructed orbit was assessed based on the postoperative CT scan.RESULTS AND CONCLUSION: Postoperative CT scans showed that the implanted individualized titanium meshes were capable of accurately reconstructing the fractured orbit in all the 12 patients, and there was no infection, and titanium mesh loosening, prolapse, and rejection. With healthy eyes as controls, 11 cases of eyeball retraction were corrected completely, and only 1 case was still under correction. Diplopia symptoms disappeared in the 8 of 9 cases, and relieved in the 1 of 9 cases. Seven cases of eye movement limitation recovered postoperatively. To conclude, the individualized titanium mesh has great accuracy to repair orbital fractures in patients without serious complications,which has achieved good clinical outcomes in the orbital reconstruction.

Ectopic osteogenesis of bone marrow stromal stem cells under bone morphogenetic protein 2/vascular endothelial growth factor 165 co-transfections / 中国组织工程研究

BACKGROUND: Double gene transfection using bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor 165 (VEGF165) for bone marrow stromal stem cells (BMSCs) to induce osteogenesis provides experimental basis for the study on tissue engineering bone. OBJECTIVE: To investigate the ability of BMP2 and VEGF165 double gene modified rat BMSCs to induce osteogenesis. METHODS: BMSCs were isolated from the femur and tibia of four 4-week-old Sprague-Dawley rats by whole bone marrow adherent culture. Passage 3 BMSCs were randomized into five groups: non-transfection group, empty plasmid group, BMP2 transfection group, VEGF165 transfection group, BMP2 and BMP2/VEGF165 transfection group (co-transfection group). Then, western blot assay was used to detect expression of BMP2 and VEGF165 at 48 hours after transfection, and the activity of alkaline phosphatase was detected in each group at 7 days after transfection. RESULTS AND CONCLUSION: Highly expressed BMP2 in BMP2 and co-transfection groups and highly expressed VEGF165 in VEGF165 and co-transfection groups were found after transfection. The expression of BMP2 or VEGF165 in the co-transfection group was significantly higher than that in the BMP2 or VEGF165 transfection group after transfection, respectively (P BMP2 transfection group > VEGF165 transfection group > empty plasmid group and non-transfection group. There was a significant difference in the activity of alkaline phosphatase between co-transfection group and any of single gene transfection groups (P < 0.05). To conclude, BMP2/VEGF165 co-transfection promotes the ectopic osteogenesis of BMSCs.

The expression of 5-hydroxytryptamine receptor 2B in gastric cancer and its clinical significance / 中华普通外科杂志

Objective To explore the expression and distribution characteristics of 5-hydroxytryptamine receptor 2B (5-HT2BR) in tumor tissue,blood and gastric juice of patients with gastric cancer.Methods From 2006 to 2013 gastric cancer tissue paraffin specimens were collectec from 30 stomach cancer patients,gastric ulcer paraffin specimens from 10 patients,and gastric juice,blood samples and fresh tumor tissues were collected from gastric cancer patients undergoing surgery.Corresponding samples were collected in 15 cases of chronic superficial gastritis while receiving gastroscopy.By using ELISA and PV-9000 immunohistochemistry 5-HT2BR expression was determined.Results There was no positive expression of 5-HT2BR in benign gastric tissues (n =15).The positive expression rate of 5-HT2BR between gastric cancer and gastric benign lesions has significant difference (P＜0.01).Integral optical density value and positive area of 5-HT2BR in low differentiation tissues of gastric cancer were higher than high and medium differentiation tissues of gastric cancer (P ＜ 0.01).There was no statistically significant difference among 5 HT2BR and constituent ratio of patients at tumor stage,differentiated degree,histological type,infiltration depth and lymph node metastasis (all P ＞ 0.05).The differences of 5-HT2BR expression in gastric juice and blood between gastric cancer patients and chronic gastritis patients are statistically significant(P ＜ 0.01).Conclusions 5-HT2BR are highly expressioned in blood,gastric juice and tumor tissues of patients with gastric cancer in a differentiation related manner.

Detection of the Zaire Subtype of the Ebola Virus by Isothermal Multiple Self-matching Initiated Amplification / 病毒学报

Given the Ebola outbreak in West Africa and the risks of spread to other regions, a rapid, sensitive and simple method for the detection of the Ebola virus (EBOV) is of great significance for the prevention and control of Ebola. We developed a simple colorimetric isothermal multiple self-matching initiated amplification (IMSA) for rapid detection of the Zaire subtype of the Ebola virus (EBOV-Z). This method employed six primers that recognized seven sites of the EBOV-Z nucleoprotein gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 1 h. Amplification products were detected through visual inspection of color change by pre-addition of hydroxyl naphthol blue dye. Relative sensitivity was validated by detection of serial tenfold dilutions of virus-like particles containing the partial EBOV-Z nucleoprotein gene and mock clinical sample. Specificity of IMSA was validated by detection of the plasma of 30 healthy volunteers, the dengue virus, and Japanese encephalitis virus. IMSA had comparable sensitivity to Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and cross-reaction with human plasma or other viruses was not observed. Reverse transcription-isothermal multiple self-matching initiated amplification (RT-IMSA) was also evaluated and compared in parallel with the commercial RT-qPCR kit for detection of EBOV-suspected samples of human blood in Sierra Leone. Sensitivity and specificity of the RT-IMSA was 91.4% and 100%, respectively. These data suggest that RT-IMSA is a valuable tool for the detection of the EBOV with the distinct advantages of simplicity and low cost compared with RT-qPCR.

Reconstruction of the orbital fracture with enophthalmos using customized titanium mesh combined with Medpor / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aims to evaluate the effect of customized titanium mesh, which was designed by mirror imaging and rapid prototyping technique, with Medpor applied for orbital fracture with enophthalmos.</p><p><b>METHODS</b>Orbital axial, coronal, and sagittal CT scan, and three-dimension CT examination were performed routinely in. 18 cases of orbital fracture with enophthalmos preoperatively. Based on the CT data, prosthesis model was designed by reverse engineering and rapid prototyping technique. The customized titanium mesh and Medpor were applied for orbital reconstruction and enophthalmos correction. Orbital reduction and globe projection were evaluated by postoperative CT scan and clinical follow-up visits.</p><p><b>RESULTS</b>CT scans revealed that the customized titanium mesh with Medpor had great accuracy to reconstruct the orbital fracture and correct the enophthalmos in all patients without serious complications. The eye and facial appearance and function recovered significantly. No serious complication occurred in the operation and after operation.</p><p><b>CONCLUSION</b>The customized titanium mesh, based on mirror imaging and rapid prototyping technique, can accurately reconstruct the orbital fractures with enophthalmos. The application of Medpor can effectively correct enophthalmos and eyeball mobility malformation.</p>

Application of anterolateral thigh myocutaneous flap using computed tomography angiography for mouth-floor reconstruction after resection of middle-late stage carcinoma of mouth floor / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The aim of this study was to investigate the value of free anterolateral thigh myocutaneous flap (ALTMF) and computed tomography angiography (CTA) for the reconstruction of mouth-floor defects after the resection of middle-late stage carcinoma of the mouth floor.</p><p><b>METHODS</b>Sixteen cases of middle-late stage carcinomas of the mouth floor underwent radical resection, and mouth-floor and tongue defects were reconstructed with ALTMF. CTA was applied to plan the lateral circumflex femoral artery (LCFA) and its perforating vessel, which was verified during the operation.</p><p><b>RESULTS</b>The position of the perforating vessel in the operation was fully consistent with that designed by the preoperative CTA. All 16 flaps completely survived. The appearance and function of all cases were both satisfactory. All donor sites were primarily closed and healed without functional morbidity. During the follow-up period of 6-36 months, 15 cases survived with acceptable aesthetic and functional results in mouth floor and tongue reconstruction, except for 1 case (T4N2M0) that died of metastasis carcinoma 10 months after operation.</p><p><b>CONCLUSION</b>CTA can accurately locate the LCFA and artery perforator. Preoperative perforator planning using CTA in ALTMF transplantation is a reliable and useful method thatresults in safe operation with optimal outcome. The ALTMF is an ideal choice for the reconstruction of soft tissue defects after the resection of middle-late staie carcinoma of the mouth floor</p>

Efficacy of intravenous methylprednisolone for prevention of tracheal intubation-related laryngopharyngeal complications / 中华麻醉学杂志

Objective To evaluate the efficacy of intravenous methylprednisolone for prevention of tracheal intubation-related laryngopharyngeal complications.Methods Three hundred ASA Ⅰ or Ⅱ patients,aged 20-50 yr,weighing 50-80 kg,undergoing elective surgeries,requiring tracheal intubation under general anesthesia,were included and randomized into 5 groups (n =60 each).Methylprednisolone 40 and 80 mg were injected intravenously at 30 min before induction of anesthesia in groups Ⅰ and Ⅱ,respectively,while the equal volume of normal saline was given instead in group Ⅲ.Methylprednisolone 40 and 80 mg were injected intravenously at 30 min before extubation in groups Ⅳ and Ⅴ,respectively.The sore throat,hoarseness and cough were recorded within 24 h after extubation and the severity was evaluated at 1 and 24 h after extubation.Results There was no significant difference in the incidence and severity of sore throat,hoarseness and cough between the five groups (P ＞ 0.05).Conclusion Intravenous methylprednisolone can not effectively prevent tracheal intubation-related laryngopharyngeal complications in patients.

Prominent zygomatic complex reduction by C-type osteotomy / 中华医学美学美容杂志

Objective To explore the operation of C-type osteotomy for reduction of prominent zygomatic complex. Methods Based on the severity and characteristics of prominent zygomaitc complex, Ctype osteotomy was designed for the malar complex reduction by using oral and minor pre-auricular approaches under general anaesthesia. Two paralleled osteotomic lines of C-type were marked from zygomatic alveola to the conjunction of lateral orbital margin and zygomatic arch through the inferio-lateral edge of orbit. The extension of zygomatic arch reduction was determined the width of two osteotomic lines. The bone which marked lines was removed by reciprocating saw and osteotome. The zygomatic arch root was osteotomiced by pre-auricular approaches. Then, the zygomatic complex could move freely towards superior-medial position. Finally, the zygoma was fixed with titanium mini-plates. Results 12 patients with prominent zygomatic complex had been successfully operated by C-type osteotomy from July 2006 to April 2009. Of them, six cases were symmetrical and six cases were unsymmetrical. Postoperative follow-up for 4-24 months, infection was not occurred, and the scar of pre-auricular incision was not obvious. All the patients obtained positive results. Conclusion C-type osteotomy for correction of prominent zygomtic complex through intra-oral and minor pre-auricular approach is an effective surgical method and gives superior results. It preserves the intactness of maxillary sinus, prevents facial slack, and is especially effective for patients with prominent zygomatic arch.