Gyrovirus galga1(GyVg1)and gyrovirus homsa1(GyH1)are two newly discovered cir-coviruses that can cause symptoms related to transmissible viral proventriculitis of chickens.These viruses have been reported in various regions worldwide.This research aims to establish a duplex real-time PCR assay capable of identifying and detecting GyVg1 and GyH1.Specific primers and probes were designed based on the conserved regions of GyVg1 and GyH1 respectively using all genome sequence data currently available in GenBank.After optimizing reaction conditions,the du-plex real-time PCR detection method was established and further validated by comparing it with a conventional PCR assay and sequencing results from an analysis of 256 clinical samples collected in 2023 across eight regions of Nanning,Guangxi.The results showed that GyVg1 and GyH1 could be identified in 1 h by the duplex real-time PCR assay and two pairs of primer probes can amplify effectively but there is no any cross reaction with other pathogens.Besides,the detection limit was determined to be 7.5 copies/μL.The correlation coefficient of standard curves exceeded 0.99,and CV for intra-and inter-assay was less than 0.45％.Based on clinical performance,when the quanti-ty of template was greater than or equal to 100 copies,the agreements between the duplex real-time PCR assay and the conventional PCR assay were 94.3％(GyVg1)and 100％(GyH1).In con-clusion,the newly developed duplex real-time PCR assays exhibited good specificity,sensitivity and repeatability,which could contribute to the rapid detection and differentiation of GyVg1 and GyH1.

To establish a double fluorescent RT-LAMP that can distinguish between chicken astro-virus(CAstV)and avian nephritis virus(ANV),two sets of specific primers and probes labeled with fluorescent groups were designed by comparing the conserved sequences of CAstV ORF1b gene and ANV ORF1b gene downloaded from GenBank.The CAstV probe was labeled with CY3 fluorophore at the 3'end and the quencher BHQ2 at the 5'end.The ANV probe was labeled with CY3 fluorophore at the 3'end and the quencher BHQ2 at the 5'end.The reaction system of the method was optimized,and specificity,repeatability,interference,sensitivity,and clinical sample detection were performed.The results were compared with RT-qPCR and RT-PCR to verify the ef-fectiveness of the method.The experimental results showed that the established double fluorescence RT-LAMP method could complete the reaction within 60 min,and the optimal reac-tion temperature was 65 ℃.This method only detects ANV and CAstV,with no cross-reactivity to other common avian pathogens.The minimum detection limit of CAstV was 1.4 ×10 2 copies/μL,and the minimum detection limit of ANV was 1.5×10 2 copies/μL.The interference and reproduc-ibility tests were good.Compared with RT-qPCR,the coincidence rates of CAstV and ANV were 97.97％and 98.85％,respectively.In conclusion,the double fluorescence RT-LAMP method estab-lished in this study has the characteristics of rapidity,accuracy,good specificity,high sensitivity and good reproducibility,which is suitable for clinical diagnosis and provides technical support for the epidemiological investigation of CAstV and ANV.

Gyrovirus galga1(GyVg1)and gyrovirus homsa1(GyH1)are two newly discovered cir-coviruses that can cause symptoms related to transmissible viral proventriculitis of chickens.These viruses have been reported in various regions worldwide.This research aims to establish a duplex real-time PCR assay capable of identifying and detecting GyVg1 and GyH1.Specific primers and probes were designed based on the conserved regions of GyVg1 and GyH1 respectively using all genome sequence data currently available in GenBank.After optimizing reaction conditions,the du-plex real-time PCR detection method was established and further validated by comparing it with a conventional PCR assay and sequencing results from an analysis of 256 clinical samples collected in 2023 across eight regions of Nanning,Guangxi.The results showed that GyVg1 and GyH1 could be identified in 1 h by the duplex real-time PCR assay and two pairs of primer probes can amplify effectively but there is no any cross reaction with other pathogens.Besides,the detection limit was determined to be 7.5 copies/μL.The correlation coefficient of standard curves exceeded 0.99,and CV for intra-and inter-assay was less than 0.45％.Based on clinical performance,when the quanti-ty of template was greater than or equal to 100 copies,the agreements between the duplex real-time PCR assay and the conventional PCR assay were 94.3％(GyVg1)and 100％(GyH1).In con-clusion,the newly developed duplex real-time PCR assays exhibited good specificity,sensitivity and repeatability,which could contribute to the rapid detection and differentiation of GyVg1 and GyH1.

To establish a double fluorescent RT-LAMP that can distinguish between chicken astro-virus(CAstV)and avian nephritis virus(ANV),two sets of specific primers and probes labeled with fluorescent groups were designed by comparing the conserved sequences of CAstV ORF1b gene and ANV ORF1b gene downloaded from GenBank.The CAstV probe was labeled with CY3 fluorophore at the 3'end and the quencher BHQ2 at the 5'end.The ANV probe was labeled with CY3 fluorophore at the 3'end and the quencher BHQ2 at the 5'end.The reaction system of the method was optimized,and specificity,repeatability,interference,sensitivity,and clinical sample detection were performed.The results were compared with RT-qPCR and RT-PCR to verify the ef-fectiveness of the method.The experimental results showed that the established double fluorescence RT-LAMP method could complete the reaction within 60 min,and the optimal reac-tion temperature was 65 ℃.This method only detects ANV and CAstV,with no cross-reactivity to other common avian pathogens.The minimum detection limit of CAstV was 1.4 ×10 2 copies/μL,and the minimum detection limit of ANV was 1.5×10 2 copies/μL.The interference and reproduc-ibility tests were good.Compared with RT-qPCR,the coincidence rates of CAstV and ANV were 97.97％and 98.85％,respectively.In conclusion,the double fluorescence RT-LAMP method estab-lished in this study has the characteristics of rapidity,accuracy,good specificity,high sensitivity and good reproducibility,which is suitable for clinical diagnosis and provides technical support for the epidemiological investigation of CAstV and ANV.

BACKGROUND: There is no report on the treatment of suppurative discitis with posterior microendoscopic discectomy, antibiotic artificial bone implantation and one-stage percutaneous pedicle screw fixation. OBJECTIVE: To evaluate the clinical effectiveness of posterior microendoscopic debridement, antibiotic artificial bone implantation and one-stage percutaneous pedicle screw fixation for pyogenic spondylodiscitis. METHODS: Thirty-one patients with suppurative discitis admitted at the Department of Spinal Surgery, the First Affiliated Hospital of Guangzhou Medical University from January 2014 to December 2017, including 17 males and 14 females, aged 28-78 years, were included. All the patients underwent posterior microendoscopic lesion clearance and antibiotic artificial bone grafting and one-stage percutaneous pedicle screw fixation. The erythrocyte sedimentation rate and C-reactive protein levels were detected at postoperative follow-up. The Visual Analog Scale and the Japanese Orthopaedic Association scores were used for detection. RESULTS AND CONCLUSION: (1) All the surgical operations of the 31 patients were successfully completed and all patients had immediate relief of low back pain and lower extremity radiation pain. They were able to move ground wearing a waistband at 1-4 days postoperatively. (2) Thirty-one patients were followed up for 9-18 months. All patients had a significant decrease in C-reactive protein and erythrocyte sedimentation rate at 1 week after surgery, and C-reactive protein and erythrocyte sedimentation rate were in the normal range at 1, 3, and 6 months postoperatively. (3) The Visual Analog Scale scores of all patients at 1 week of follow-up were significantly lower than those before surgery (P < 0.05) , and the Japanese Orthopaedic Association scores were significantly higher than those before surgery (P < 0.05) . (4) In the follow-up examination of CT and MRI, there was no recurrence, pseudoarticular formation or internal fixation loosening. (5) These results suggest that posterior microendoscopic discectomy, antibiotic artificial bone implantation combined with one-stage percutaneous pedicle screw fixation for treating suppurative discitis can result in little trauma, few bleeding and rapid pain relief, and patients can move to the ground early. The operation method has a good clinical effect.

Objective To evaluate the accuracy and safety of pedicle-scope assisted pedicle screw fixation and percutaneous pedicle screw fixation for treating lumbar degenerative disease(LDD).Methods Thirty cases of LDD treated by mini-invasive transforaminal lumbar interbody fusion(TLIF) under microscope plus percutaneous pedicle screw fixation with fluoroscopy or pedicle-scope of lumbar spine from December 2013 to September 2015 were selected and divide into the percutaneous group and pediclescope group,15 cases in each group.The operative duration,intraoperative blood loss volume,hospital stay and complications were compared between the two groups.The clinical effects were assessed with visual analogue scale(VAS) and Oswestry disability index(ODI).The plain radiography,dynamic radiology and lumbar magnetic resonance imaging(MRI) and computed tomography (CT) were re-examined after operation for assessing the screw location and fusion rate.Results Eighty-four screws were placed in the percutaneous group and 70 screws in the pedicle-scope group.In the percutaneous group and pedicle-scope group,the mean intraoperative blood loss volume,mean operative duration,average times of C-arm fluoroscopy,mean length of hospital stay,mean time of off-bed and complication occurrence rate were measured,and the differences were not statistically significant (P＞ 0.05).In 6-month follow up,the VAS score and ODI score in the two groups were significantly improved compared with before operation(P＜0.01);the VAS score and ODI score were(2.50 ± 1.30) and (50.00-±-3.50) in pediclescope group,(3.00± 1.50) and (58.00 ±4.50) in percutaneous group,respectivehy,and the differences were statistically significant(P＜0.05);the improvement rates of clinical symptoms in the percutaneous group and pedicle-scope group were 73.33 ％ and 80.00 ％ respectively.The imaging reexamination showed that the placed screw location was good and lesion segment fusion was good without screw loosening.The location of 3 screws in the percutaneous group was deviated and adjusted by the second operation.Conclusion The accuracy and safety of pedicle-scope assisted screw placing for treating LDD are higher than those of conventional percutaneous pedicle screw fixation,moreover the operative time is shorter with less intraoperative bleeding.

BACKGROUND:The existing bone grafting apparatus are al long tubular-shape, antegrade bone grafting; however, the bone grains within the tube were difficult to put through, along with large bone loss. OBJECTIVE:To design a new concept of minimaly invasive bone grafting apparatus, so as to achieve the minimaly invasive bone grafting in vertebral body and intervertebral space. METHODS: On Solidworks 2012 software platform, the 3D three-dimensional solid modeling technology was employed to design a new concept of gun-shape bone grafting device with the characteristics of headend reversely placing at the grafting position and antegrade pushout, against the problems such as antegrade bone grafting, long bone grafting path and difficulty in bone grafting existing in precious percutaneous or/and transpedicular bone graft apparatus. The virtual assembly and simulative bone grafting surgery were performed with the bone grafting apparatus for observation of the instrument matching degree and grafting surgical procedures. RESULTS AND CONCLUSION: A set of minimaly invasive gun-shape bone grafting apparatus was successfuly designed; its length was 20 cm in total; the length of the front semi-closed bone tube was 2.5 cm. There are two kinds of diameter specifications: inner diameter of 3.5 mm/outer diameter of 4 mm, and inner diameter of 5 mm/outer diameter of 6 mm. Virtual assembly revealed that the grafting apparatus were wel matched. Stimulative surgery displayed that the bone grain loading and launch were simple. The gun-shape bone grafting device is reasonable in design, avoiding pushing too long distance, bone pushing difficulties, bone loss and other problems in previous tubular bone grafting apparatus. Percutaneous, endoscopic vertebrae or intervertebral space minimaly invasive bone grafting can be performed with this apparatus.

Objective To explore the effect of one-piece shoulder pillow on patients suffering from shoulder pain and skin erubescence during gynecological laparoscopy with lithotomy position and trendelenburg position. Methods Two hundred and six patients who underwent gynecological laparoscopic operation were divided into control and experiment groups according to cardinal or even number of registration. The patients in both groups were set in lithotomy position and trendelenburg position during the operation, with the difference in the use of a common shoulder pillow for support in the control group but no use in the experiment one. On-position time and incidence rate of shoulder pain and skin erubescence were compared between two groups. Result The sholder pain and skin erubescence were less than those of control group (P < 0.05). Conclusion Use of one-piece shoulder pillow can reduce the incidence rates of shoulder pain and skin erubescence after gynecological laparoscopic operation with lithotomy position and trendelenburg position.

BACKGROUND: Magnetic resonance imaging(MRI)is considered safe for diagnosis of meniscus injury,having accuracy as high as 80% to 100%.However,it is found that there are some false positive and false negative in the clinic.OBJECTIVE: Through assessment of menisci injuries of the knee using magnetic resonance imaging and arthroscopy,to investigate MRI manifestation in different types of meniscal injuries.METHODS: A study from January 2003 to December 2008 was performed in 212 patients with knee injury undergoing preoperative MRI.Radiograph was read by physicians who had a senior professional title from radiology and orthopaedics departments,and meniscus damage and damage morphology were predicted.Arthroscopy was conducted by chief physician.Meniscus damage during surgery was considered as gold standards.The accuracy of them was compared.The statistics of accuracy of MRI diagnosis was gathered.RESULTS AND CONCLUSION: Totally 212 patients and 230 knees were included,and a total of 215 knees of meniscus injury were described by MRI diagnosis and 213 knees of meniscus injury were proven by arthroscopy.The sensitivity,specificity and accuracy of MRI diagnosis of meniscus injury respectively were 93.5%,64.7%,93.0%.It is found that the sensitivity of MRI diagnosis of bucket-handle damage is high.MRI is easy to miss diagnosis in a light meniscus posterior horn tear,and resuts are more false positive in lateral meniscus injury.It needs arthroscope operation or further surgery in some condition such as gradeⅢ type injury,meniscus displacement after injury and articular capsule separation,which indicates instable meniscus.