This study was aimed at investigating the effectiveness of various host nucleic acid removal and non-specific amplifica-tion techniques in animal tissue samples,to increase the accuracy of pathogen identification in tissue samples.Simulated samples were prepared with a mixture of mouse lung tissue homogenates and Klebsiella pneumoniae fluids,and processed with six host nucleic acid removal kits and three non-specific amplification techniques.The effectiveness of each method in removing host DNA and enriching nucleic acids of pathogenic microorganisms was evaluated through real-time fluorescence quantitative PCR and high-throughput se-quencing.For host nucleic acid removal techniques,the method of selective cleavage and quantitative degradation of host DNA(Com-plete5 kit)effectively decreased the host nucleic acid content in tissue samples and increased the relative abundance of pathogen nucleic acids.In contrast,the magnetic bead method for host DNA removal(Next microbiome DNA enrichment Kit kit)was less effec-tive.At lower pathogen concentrations(77 CFU/mL),the Vazyme kit was more effective than the other kits in removing host nucleic acids.Non-specific amplification techniques(MALBAC whole genome amplification,MDA isothermal amplification,and random primer amplification)were not applicable to tissue samples and were not effective in increasing the relative abundance of pathogen nucleic acids.Selective lysis and quantitative degradation of host DNA were suitable for processing tissue samples with high host back-ground and low pathogenic microorganism levels,whereas non-specific amplification methods were not applicable to tissue samples for pre-processing of macro-genome high-throughput sequencing.

This study was aimed at investigating the effectiveness of various host nucleic acid removal and non-specific amplifica-tion techniques in animal tissue samples,to increase the accuracy of pathogen identification in tissue samples.Simulated samples were prepared with a mixture of mouse lung tissue homogenates and Klebsiella pneumoniae fluids,and processed with six host nucleic acid removal kits and three non-specific amplification techniques.The effectiveness of each method in removing host DNA and enriching nucleic acids of pathogenic microorganisms was evaluated through real-time fluorescence quantitative PCR and high-throughput se-quencing.For host nucleic acid removal techniques,the method of selective cleavage and quantitative degradation of host DNA(Com-plete5 kit)effectively decreased the host nucleic acid content in tissue samples and increased the relative abundance of pathogen nucleic acids.In contrast,the magnetic bead method for host DNA removal(Next microbiome DNA enrichment Kit kit)was less effec-tive.At lower pathogen concentrations(77 CFU/mL),the Vazyme kit was more effective than the other kits in removing host nucleic acids.Non-specific amplification techniques(MALBAC whole genome amplification,MDA isothermal amplification,and random primer amplification)were not applicable to tissue samples and were not effective in increasing the relative abundance of pathogen nucleic acids.Selective lysis and quantitative degradation of host DNA were suitable for processing tissue samples with high host back-ground and low pathogenic microorganism levels,whereas non-specific amplification methods were not applicable to tissue samples for pre-processing of macro-genome high-throughput sequencing.

The risk stratification of acute pulmonary thromboembolism(PTE)is the direction of treatment,and corresponding treatment plans are adopted for patients with different risk stratification.Anticoagulant therapy is the cornerstone of the treatment of acute PTE,but simple anticoagulant therapy is associated with an increased risk of early hemodynamic decompensation and death in the medium to high-risk population.Systematic thrombolysis can quickly reduce the burden of thrombus,alleviate right ventricular dysfunction,and improve hemodynamics earlier,but there is a high risk of bleeding.At present,catheter interventional therapy is rapidly developing and many matching devices have been developed,which aims to better formulate the treatment plan for acute PTE and to better meet the clinical requirements for the treatment of PTE.This article,based on the therapeutic idea of risk stratification,aims to make a comprehensive review about the different types of mechanical thrombectomy devices,inhalation thrombectomy devices and lytic thrombectomy devices,which are currently used in clinical treatment of PTE,although a comparative study on the advantages of their clinical application has not been conducted yet.The advantages and trends of their respective applications are discussed so as to provide a systematic summary of the current academic research in this field.

Objective:To investigate the clinical efficacy of double tract reconstruction of proximal gastric resection and Roux-en-Y total gastrectomy in the treatment of adenocarcinoma of the esophagogastric junction, in order to provide reference for clinical selection.Methods:From June 2016 to June 2018, patients with adenocarcinoma of the esophagogastric junction in Chifeng City hospital were prospectively selected and randomly divided into proximal gastrectomy double channel anastomosis group (PG group) and total gastrectomy Roux-en-Y anastomosis group (TG group). The perioperative indexes, clinical pathology, early (≤ 30 days) and late (> 30 days) complications of the two groups were compared, and the nutritional status [body mass index (BMI), prealbumin, albumin, total protein], and anemia degree [hemoglobin (Hb), red blood cell count (RBC)] of the two groups at 3 months, 6 months and 12 months after operation were compared.Results:A total of 83 patients with adenocarcinoma of the esophagogastric junction were admitted, including 62 patients who were eligible for surgery, 31 cases in each group, 1 case in each group was lost to follow-up, and 30 cases in each group. There were no tumor recurrence or death in two groups. There were no statistically significant differences in age, gender, tumor size, tumor stage, preoperative complications, preoperative nutritional status and anemia between the two groups ( P>0.05). The time of first exhaust, time of starting fluid feeding and postoperative hospitalization in PG group was significantly earlier than that in TG group [(3.0±0.7)days vs (4.2±0.7)days, (7.9±0.9)days vs (9.0±0.9)days, (13.3±1.1)days vs (14.6±0.9)days, P<0.05], while the operation time was slightly longer [(252.0±36.9)min vs (225.8±31.1)min, P<0.05]. There was no significant difference in intraoperative blood loss and number of lymph node dissection between PG group and TG group [(241.7±81.0)ml vs (245.8±71.9)ml, (40.5±12.2)nodes vs (43.2±10.3)nodes, P>0.05]. There was no statistically significant difference in the occurrence of early and late postoperative complications ( P>0.05). The incidence of reflux esophagitis in PG group was less than that in TG group ( P<0.05). The nutritional status of PG group was significantly better than TG group in 3 months, 6 months and 12 months after the operation ( P<0.05). The Hb and RBC decreased in both groups at 3 and 6 months after operation, and the Hb and RBC in PG group were higher than those in TG group. There was no anemia in PG group and TG group 12 months after operation. Conclusions:Double tract reconstruction of proximal gastric resection is effective in the treatment of adenocarcinoma of the esophagogastric junction. It is worthy of further clinical promotion.

The fcl gene encodes GDP-fucose synthase, which catalyzes two-step differential isomerase and reductase reactions in the synthesis of GDP-L-fucose from GDP-D-mannose. It also participates in the biosynthesis of amino sugar and ribose sugar, and is one of the key enzymes to regulate the metabolism of sugar and nucleotides in organisms. The presence of fcl gene in Saccharopolyspora pogona was found through sequencing result of genome. The mutant S. pogona-fcl and S. pogona-Δfcl were constructed by gene engineering technology. The results showed that the gene had an effects on growth and development, protein expression and transcriptional level, insecticidal activity, and biosynthesis of butenyl-spinosyn of Saccharopolyspora pogona. The results of HPLC analysis showed that the yield of butenyl-spinosyn in S. pogona-Δfcl was 130% compared with that in S. pogona, which reduced by 25% in S. pogona-fcl. The results of determination of insecticidal activity showed that S. pogona-Δfcl had a stronger insecticidal activity against Helicoverpa armigera than that of S. pogona, while the S. pogona-fcl had a lower insecticidal activity against Helicoverpa armigera compared with S. pogona. Scanning electron microscopy (SEM) was used to observe the morphology of the mycelia. It was found that the surface of the S. pogona-Δfcl was wrinkled, and the mycelium showed a short rod shape. There was no significant difference in mycelial morphology between S. pogona-fcl and S. pogona. Aboved all showed that deletion of fcl gene in S. pogona hindered the growth and development of mycelia, but was beneficial to increase the biosynthesis of butenyl-spinosyn and improve insecticidal activity. Whereas the fcl gene over-expression was not conducive to the biosynthesis of butenyl-spinosyn and reduced their insecticidal activity. SDS-PAGE results showed that the difference of protein expression among the three strains was most obvious at 96 hours, which was identified by real-time fluorescence quantitative polymerase chain reaction, the results showed that there were significant differences of related genes in transcriptional levels among the three strains. Based on the results of the study, a network metabolic control map was constructed to analyze the effect of fcl gene on growth and the regulation pathway of butenyl-spinosyn biosynthesis, which provided an experimental basis for revealing the regulation mechanism of butenyl-spinosyn biosynthesis and related follow-up studies.
Bacterial Proteins ; Genetic Engineering ; Insecticides ; Macrolides ; Saccharopolyspora

Objective To discuss the application of shear wave elastography and strain elasticity imaging diagnosis of infants with congenital muscular torticollis. Methods Children with congenital muscular torticollis in 50 patients and 100 healthy infants were recruited for the research at Shezhen Children′s Hospital in the period of September to December, analyzing factors in normal sternocleidomastoid two mode ultrasonic elastography, including the young′s modulus, shear wave velocity, shear modulus and the ratio of strain results, comparing the change of elasticity between the patients and normal. Results The elastic parameters of the sternocleidomastoid showed no statistically significant difference between the genders, the left side and the right side. The results of young′s modulus, shear wave velocity and shear wave modulus were higher in long axis section than the short axis section, and the difference was statistically significant (P<0.05). However, the results of the strain ratio and thickness was not statistically significant. The young′s modulus, shear wave velocity and shear wave modulus are higher in the stretch back position than symmetrical supine position, and the difference is statistically significant (P<0.05). But those of strain ratio and thickness was not statistically significant. The thickness of the sternocleidomastoid muscle and the young′s modulus, shear wave velocity, shear modulus and the ratio of strain results ultrasound elastic results were higher in the lesion side than the healthy side, and the difference is statistically significant (P<0.05). Conclusion The ultrasound elastic imaging of multimodal technique can quantify the elasticity of the normal infant and the patients sternocleidomastoid, be used for diagnose for the infants with the congenital muscular torticollis.

Objective To analysis the change of hip joint in healthy infants by ultrasound,and establish the normal reference value of the developmental dysplasia of the hip(DDH). Methods A total of 8 000 healthy infants from 0 to 24 weeks were collected from the Multi-center study of 10 children′s medical centers. Among them,3 855 infants(2 065 females and 1 790 males) with complete data and follow-up were included in this study. All subjects were divided into 6 groups ( ＜4,4～7,8～11,12～15,16～19 and≥20 weeks group). α angle,femoral head length and width,femoral head coverage ratio by acetabulum ( FHC) were measured in the coronal view on the neutral position;distance from pubis to femoral head ( P-H) and distance from ischium to femoral head ( I-H ) were measured in the transverse view on neutral position;distance from femoral head topubis ( H-P) was measured in the posterolateraltransverse view on the flexion position. The results of each group changes with age were analysised. Results ① The α angle of healthy infants from 0 to 20 weeks were increased with age,the difference among the groups were statistically significant( P ＜0.05),but there was no significant difference between 16～19 and ≥20 weeks group( P ＞0.05). ②The femoral head length and width of all age groups were increased with age,the difference among all the groups was statistically significant( all P ＜0.05). ③ FHC from 0 to 20 weeks were increased with age,the difference among the groups were statistically significant( P ＜0.05) except between 16～19 and ≥20 weeks group( P ＞0.05). ④ The P-H and I-H in all age groups showed no statistically significant ( all P＞0.05). ⑤The H-P of all age groups were increased with age,the difference between the groups were statistically significant(all P ＜0.05).Conclusions The development of hip joints have the certain regular developmental pattern in healthy infants less than 5 months of birth and are relatively constant after birth more than 5 months. The ultrasound normal reference value of the hip joints can be used for the early diagnosis of the DDH.

Objective: To investigate reduction and fixation of complex acetabular fractures using three-dimensional (3D) printing technique and personalized acetabular wing-plate via lateral-rectus approach. Methods: From March to July 2016, 8 patients with complex acetabular fractures were surgically managed through 3D printing personalized acetabular wing-plate via lateral-rectus approach at Department of Orthopedics, the Third Affiliated Hospital of Southern Medical University. There were 4 male patients and 4 female patients, with an average age of 57 years (ranging from 31 to 76 years). According to Letournel-Judet classification, there were 2 anterior+ posterior hemitransverse fractures and 6 both-column fractures, without posterior wall fracture or contralateral pelvic fracture. The CT data files of acetabular fracture were imported into the computer and 3D printing technique was used to print the fractures models after reduction by digital orthopedic technique. The acetabular wing-plate was designed and printed with titanium. All fractures were treated via the lateral-rectus approach in a horizontal position after general anesthesia. The anterior column and the quadrilateral surface fractures were fixed by 3D printing personalized acetabular wing-plate, and the posterior column fractures were reduction and fixed by antegrade lag screws under direct vision. Results: All the 8 cases underwent the operation successfully. Postoperative X-ray and CT examination showed excellent or good reduction of anterior and posterior column, without any operation complications. Only 1 case with 75 years old was found screw loosening in the pubic bone with osteoporosis after 1 month′s follow-up, who didn′t accept any treatment because the patient didn′t feel discomfort. According to the Matta radiological evaluation, the reduction of the acetabular fracture was rated as excellent in 3 cases, good in 4 cases and fair in 1 case. All patients were followed up for 3 to 6 months and all patients had achieved bone union. According to the modified Merle D′Aubigné and Postel scoring system, 5 cases were excellent, 2 cases were good, 1 case was fair. Conclusions Surgical management of complex acetabular fracture via lateral-rectus approach combine with 3D printing personalized acetabular wing-plate can effectively improve reduction quality and fixation effect. It will be truly accurate, personalized and minimally invasive.

Objective To investigate the diagnostic value of ultrasonography in radial head subluxation (RHS).Methods The clinical data and ultrasonographic findings of 34 children with RHS were retrospectively analyzed.All patients were routinely treated by bilateral compared and multi-sectional elbow joints ultrasonography.And the X ray results were negative.Results Ultrasonographic manifestations of RHS included increased radiocapitellar distance in 29 cases (29/34,85.29％);widened joint space and enhanced echo in 25 cases (25/34,73.53％);hook sign,supinator muscle above the radial head in 32 cases (32/34,94.12 ％);annular ligament entrapment in 33 cases (33/34,97.06％).Conclusion Ultra sonographic manifestations of RHS have certain characteristics.Comparation of bilateral elbow joints and the application of continuous scanning ultrasound are helpful to make a quick and accurate diagnosis of X-ray negative RHS.

Objective:To investigate the effects of icaritin(ICT)on the proliferation and osteogenic differentiation of rat bone mar-row stromal cells(rBMSCs).Methods:rBMSCs were cultured from the bone marrow of SD rats and identified by multilineage differ-entiation assays.3,6 and 9 days after the treatment of rBMSCs of passage 4 by ICT at 1 0 -9 ,1 0 -8 ,1 0 -7 ,1 0 -6 and 1 0 -5 mol/L re-spectively,the proliferation and differentiation of the cells were examined by cck-8 and alkaline phosphatase (ALP)activity assay kit respectively.The calcium nodule formation was observed by alizarin red(AR)staining 21 days after 1 0 -9 mol/L ICT treatment. Results:Primary rBMSCs showed the typical spindle-like shape with attachment growth.rBMSCs could be induced to osteogenic and adipogenic differentiation.The proliferation of rBMSCs was inhibited but ALP activity was enhanced by ICT.1 0 -9 mol/L ICT in-cresed calcium nodule formation.Conclusion:ICT can dose-dependently inhibit the proliferation,but promote the osteogenic differ-entiation of rBMSCs.