Objective To investigate the relationship of gene transfection efficiency with different ultrasound exposure time and different dose of microhuhble,and to find the appropriate ultrasound parameters for gene transfection. Methods Plasmid encoding enhanced green fluorescent protein(pEGFP) was chosen as a report gene and HepG2 cells were chosen as research object. The HepG2 cells plus pEGFP and different dose of microbubble were exposed to ultrasound(1 MHz,0.5 W/cm2) with varying time. Twenty-four hours later, the expression of EGFP in the cells was observed by fluorescence microscope,the transfection efficiency was assessed by FACS and the cell viability was observed by trypan blue exclusion. Results The expression of EGFP in all experimental groups was different,and the approving transfection efficiency was got by ultrasound exposed for 20 s when the dose of microbubble was 60 μl. Conclusions With fixed ultrasound frequency and power, different transfection efficiency was got when the exposure time and dose of microbubble were different. The appropriate parameter was 20 s,60 μl, which can supply information for further study.

Objective To assess scoliosis in adolescence and adults by three-dimensional ( 3-D ) ultrasound imaging . Methods A commercial available ultrasound instrument with a magnetic sensor system was used for long distance 3-D ultrasound imaging .Specific phantoms were imaged to evaluate the precision of this imaging system . Twenty healthy adults and 20 patients with scoliosis were imaged by 3-D ultrasonography and radiography . The deformity angle of the patients with scoliosis was measured on the reconstructed coronal plane ( C plane) of 3-D ultrasound imaging . The relevance between the curvature measured by ultrasound and the Cobb angles originated by radiography were evaluated . Results There was no significant difference between the angles of the phantom measured by the 3-D ultrasound imaging system and that measured by an angle gauge ( P = 0 .479) . The transverse process ,vertebral arch and acoustic shadow of the central spinous process of spines in healthy adults can be displayed on the reconstructed C plane by 3-D ultrasound imaging . The morphology of scoliosis in all the twenty patients can be observed on the reconstructed C planes ( multi-layers) by 3-D ultrasound imaging ,which accorded well with that observed on radiograph . In 9 patients ,part of the spinal segment could not be totally displayed on one single coronal plane and no additional measurement was carried out . In 11 patients ,all the segments of the spine could be clearly displayed on a single coronal plane and the curvature of scoliosis was then measured . There were no significant differences between the angle measured on 3-D ultrasound imaging and the Cobb angle measured on radiograph( P =0 .974) and the two measurement methods have high correlation( r =0 .991 , P <0 .0001) . Conclusions The commercial available 3-D ultrasound imaging system applied in this study offers a viable method to assess scoliosis in adolescence and adults in a reliable and radiation-free manner .

Objective To evaluate the clinical efficacy and complications of lattice carbon dioxide laser in the treatment of female mild to moderate stress incontinence.Methods 30 cases of mild to moderate stress urinary incontinence in our hospital from August to October 2017 were reviewed.The average age was (46.5 ±3.5) years old,all married and has been bred,12 patients with mild stress incontinence and 18 patients with moderate stress urinary incontinence.Preoperative preparation including:1h urine cushion test measured leakage of urine,urine flow rate,residual urine,the international consultation on incontinence questionnaire (ICI-Q-SF) score,urine routine examination.Preoperative urine flow rate (31.87 ± 2.42) ml/s,preoperative (1 h) pad test (8.19 ± 2.42) ml,preoperative ICI-Q-SF score (4-8).The power was 12.5 mJ,the efficacy was 25％,and the single treatment was chosen.Urine flow rate,1 h urine pad test,residual urine test,ICI-Q-SF score,and subjective satisfaction were recorded at 1,3,6 months after operation.Results All 30 patients in this group were followed up for more than 6 months.The pad test was (2.14 ± 0.36) ml,(1.02 ± 0.54) ml,(0.80 ± 0.41) ml at 1,3 and 6 months postoperatively,which were significantly different from preoperative pad test (P ＜ 0.01,).The urine flow rate at 1,3,6 months after operation was (30.53 ± 3.15) ml/s,(32.19 ± 2.72) ml/s,(31.23 ± 5.76) ml/s,respectively,and there was no significant difference between the 3 groups (P ＞ 0.05).There was no significant difference between preoperative and postoperative urine flow rate (P ＞ 0.05).The ICI-Q-SF at 1,3,6 months after operation were 3 (1-4),3 (0-4),3 (0-4),which were significantly different from preoperative ICI-Q-SF.Among the 30 patients,3 cases had frequent urination after operation,1 case had frequent urination before operation,and became more severe after operation.The residual urine of 30 patients was negative after operation.All patients had no dysuria and no vaginal bleeding.No signs of urinary incontinence were observed after 6 months follow-up.Conclusions For patients with mild stress urinary incontinence,there is no obvious serious complication in the short term after the laser treatment of dot matrix carbon dioxide laser.The clinical treatment is safe and the leakage of urine can be improved.

Objective Budd-Chiari syndrome is apt to be misdiagnosed,so we explore its diagnosis and treatment by liver transplantation.Methods We retrospectively analyzed the clinical data of two patients who underwent liver transplantation for Budd-Chiari syndrome.One patient was misdiagnosed before the transplantation and another was diagnosed correctly.Results Both patients were grouped to Child C category with decompensated liver cirrhosis.Patient 1 was diagnosed as recurrent hepatocellular carcinoma,but the etiology of liver disease was first unknown then suspected to be schistosomiasis.This patient underwent piggyback liver transplantation.Because there was significant swelling in the perineum and lower extremities after liver transplantation,we re-reviewed the preoperative imaging data and found communicant veins between hepatic veins,which proved that the patient was actually suffered from Budd-Chiari syndrome with hepatic vein and suprahepatic vena cava occlusion before the transplantation.After conservative treatment,the swelling of the lower body was alleviated,however,the long-term survival of the patient would be compromised.Learning from the first case,we found communicant veins between hepatic veins in imaging data of patient 2,resulting in correct diagnosis of Budd-Chiari syndrome with hepatic vein and retrohepatic vena cava diseases before the transplantation,so the patient underwent orthotopic liver transplantation,in which the liver and retrohepatic vena cava were resected,and recovered uneventfully.Liver function was normal during the follow up period of 7 months.Conclusion We should consider the possibility of Budd-Chiari syndrome in patients with unexplained end-stage liver diseases.Communicant veins between the hepatic veins shown in thin CT or MRI image are the characteristic sign for diagnosing Budd-Chiari syndrome.Simultaneously hepatic vein or cava vena disease determines the choice of various technique of liver transplantation.

In recent years, although the quantity of organ donation after citizen's death has been constantly increased, a large number of patients with end-stage renal diseases are waiting for kidney transplantation every year. The imbalance between donor and recipient is still one of the main problems affecting kidney transplantation in clinical practice. Therefore, it is of clinical significance to accurately evaluate the quality of donor kidney and fully utilize the expanded criteria donor kidney. Contrast-enhanced ultrasound has been gradually applied in the detection of multiple solid organs due to its safety, portability, real-time detection, quantification and other characteristics, and it also has promising application prospect in the evaluation of donor kidney quality. In this article, the advantages and limitations of current evaluation methods for donor kidney and current status and advantages of contrast-enhanced ultrasound in donor kidney evaluation were reviewed, and the application prospect of contrast-enhanced ultrasound in the evaluation of donor kidney quality was discussed, aiming to increase the methods and enhance the accuracy for donor kidney evaluation, and provide reference for rational use of expanded criteria donor kidney.

BACKGROUND:As a dominant breed pig in southwest China,the southern Yunnan small-ear pig has been widely used as an experimental animal in the basic research of other disciplines,but there are still no reports on its application in anterior cruciate ligament reconstruction. OBJECTIVE:To establish a large animal model of the southern Yunnan small-ear pig with anterior cruciate ligament with autologous Achilles tendon was established. METHODS:Twenty adult female Yunnan small-ear pigs were equally randomized into two groups.In the autologous Achilles tendon group,the right knee anterior cruciate ligament was reconstructed with autologous Achilles tendon as a graft,while in the sham-operated group,a similar operation was performed on the right knee without any treatment of the anterior cruciate ligament.General conditions of each pig were observed and recorded before and 12 months after surgery.Ligaments and grafts were taken for gross observation and MAS scoring.Hematoxylin-eosin staining was performed to observe morphological characteristics of ligaments.The staining and arrangement of type I and type Ⅲ collagen were evaluated by immunohistochemistry.Transmission electron microscopy was used to observe the type,size,diameter,ratio,and distribution of collagen fibers in ligaments. RESULTS AND CONCLUSION:All animals had normal diet and activity,good wound healing,no obvious inflammatory reaction,no local purulent infection,and no significant changes in mental and urinary conditions compared with those before surgery.The reconstructed cruciate ligament of the knee was intact,with no stiffness and normal range of motion.Both the anterior drawer and Lachman tests were negative.Gross observation of the graft:12 months after surgery,the grafts was in good position,with good integrity,obvious tension,ligament color close to the original anterior cruciate ligament,and complete surface synovial coverage.Most of the intraarticular ligaments in the autologous Achilles tendon group were defined as MAS I type and a few were defined as MAS Ⅱ type.In the sham-operated group,the intraarticular ligament was defined as MAS I type.Hematoxylin-eosin staining indicated that,12 months after surgery,collagen fibers in the autologous Achilles tendon group began to appear bundled,isotropic,and uniformly arranged,with more obvious isotropic corrugations,and the nuclei were mainly linear or spindle-shaped,which were similar to those in normal anterior cruciate ligament tissue of the sham-operated group.Immunohistochemistry results indicated that,12 months after surgery,there was a higher expression of type I collagen and significantly less expression of type Ⅲ collagen in the reconstructed anterior cruciate ligament in the autologous Achilles tendon group.The degree of type I and type Ⅲ staining was similar in the two groups.Under the transmission electron microscope,the diameter,arrangement and density of collagen fibers in the reconstructed anterior cruciate ligament of the autologous Achilles tendon group were similar to those of the original anterior cruciate ligament at 12 months after surgery,indicating that the ligament remodeling process had been basically completed in the autologous Achilles tendon group at 12 months after surgery.Through a comprehensive evaluation of animal general conditions,ligament general view,MAS score,hematoxylin-eosin staining,immunohistochemistry,and transmission electron microscopy observation,we successfully established a large animal model of anterior cruciate ligament reconstruction using autogenous Achilles tendon in southern Yunnan small-ear pigs,with good morphological,histological and ultrastructural results.

BACKGROUND:We have successfully established an animal model of small ear pig in southern Yunnan province with internal tension relieving technique combined with autologous Achilles tendon for anterior cruciate ligament reconstruction,and verified the stability and reliability of the model.However,whether internal tension relieving technique can promote the ligamentalization process of autologous Achilles tendon graft has not been studied. OBJECTIVE:To investigate the differences in the process of ligamentalization between conventional reconstruction and internal reduction reconstruction of the anterior cruciate ligament by gross view,histology and electron microscopy. METHODS:Thirty adult female small ear pigs in southern Yunnan province were selected.Anterior cruciate ligament reconstruction was performed on the left knee joint with the ipsilateral knee Achilles tendon(n=30 in the normal group),and anterior cruciate ligament reconstruction was performed on the right knee joint with the ipsilateral knee Achilles tendon combined with the internal relaxation and enhancement system(n=30 in the relaxation group).The autogenous right forelimb was used as the control group;the anterior cruciate ligament was exposed but not severed or surgically treated.At 12,24,and 48 weeks after surgery,10 animals were sacrificed,respectively.The left and right knee joint specimens were taken for gross morphological observation to evaluate the graft morphology.MAS score was used to evaluate the excellent and good rate of the ligament at each time point.Hematoxylin-eosin staining was used to evaluate the degree of ligament graft vascularization.Collagen fibers and nuclear morphology were observed,and nuclear morphology was scored.Ultrastructural remodeling was evaluated by scanning electron microscopy and transmission electron microscopy. RESULTS AND CONCLUSION:(1)The ligament healing shape of the relaxation group was better at various time points after surgery,and the excellent and good rate of MAS score was higher(P<0.05).Moreover,the relaxation group could obtain higher ligament vascularization score(P<0.05).(2)The arrangement of collagen bundles and fiber bundles in the two groups gradually tended to be orderly,and the transverse fiber connections between collagen gradually increased and thickened,suggesting that the strength and shape degree of the grafts were gradually improved,but the ligament remodeling in the relaxation group was always faster than that in the normal group at various time points after surgery.(3)The diameter,distribution density,and arrangement degree of collagen fibers in the relaxation group were better than those in the normal group at all time points,especially in the comparison of collagen fiber diameter between and within the relaxation group(P<0.05).

Kunxian Capsule (KX) is a popular Chinese patent medicine for the treatment of rheumatoid arthritis, nephrotic syndrome, systemic lupus erythematosus, Henoch-Schönlein purpura, ankylosing spondylitis, psoriatic arthritis and eczema. However, there is scarcity of comprehensive information on the significance of KX in the clinical application and its side effects. Hence, it is aimed to provide a review of the significance of KX, with a focus on the pharmacological effects, clinical applications, and its adverse reactions. This review was based on the published literatures in PubMed, China National Knowledge Infrastructure and WanFang database. The articles were collected by two independent authors with no time limits applied until November 30, 2022. The search term includes Kunxian Capsule and/or clinical effect, pharmacology, disease, therapy, adverse effects and quality control. KX has been shown to be effective in the treatment of autoimmune arthritis by inhibiting inflammatory responses and inducing apoptosis. Many studies suggest that KX has anti-inflammatory and analgesic properties that aid in the improvement of joint functions. KX dispels wind, removes dampness, invigorates the kidneys, and promotes blood circulation, thereby curing various diseases. However, studies also suggest KX-related adverse reactions in multiple systems. Overall, this review highlights the scientific basis of KX in curing or preventing various diseases and provides novel insights for further research and clinical applications.