[Case-control study on remnant-preserving strategy for preservation and reconstruction of anterior cruciate ligament].

OBJECTIVE: To investigate and compare the clinical efficacies of remnant-preserving and remnant-non-preserving, remnant-non-preserving remnant segment preserving and remnant root preserving with anterior cruciate ligament reconstruction. METHODS: From March 2014 to December 2017, 204 patients with anterior cruciate ligament (ACL) injuries were treated by single-bundle ACL reconstruction with hamstring tendon autograft. According to the different methods of remnant preservation, the procedures were divided into remnant segment preserving group (A), remnant root preserving group (B), and remnant-non-preserving group (C). There were 37 males and 39 femalesin group A aged from 16 to 43 years old with an average of (28.80±5.41) years old. The time from injury to operation ranged from 2 to 11 weeks with an average of (3.68±1.04) weeks. In group B, there were 39 males and 25 females aged from 18 to 41 years old with an average of (28.42±5.60) years old. The time from injury to operation ranged from 2 to 10 weeks with an average of (3.36±1.68) weeks. In group C, there were 37 males and 27 females aged from 18 to 43 years old with an average of (29.10±6.11) years old. The time from injury to operation ranged from 3 to 11 weeks with an average of (3.54±1.46) weeks. The range of motion (ROM) of the knee was used to assess the range of extension and flextion of the knee at pre-operation and 24 months after operation. Lysholm score and the international knee documentation committee (IKDC) score were used to assess the knee function. The differences among three procedures were judged by comparing among the three groups at 6, 12 and 24 months postoperatively. RESULTS: All incisions got a one stage healing, and no complications, such as vascular injury, nerve damage and articular infect or the like, occurred. All the patients were followed up, and the follow-up duration of group A ranged from 24.00 to 45.96 months with a mean of (35.52±14.40) months;the follow up duration of group B ranged from 27.96 to 48.00 months with a mean of (37.56±10.68) month;and the follow up duration of group C ranged from 24.00 to 66.00 months with a mean of (37.08±13.44) month. There were no significant differences in follow up time among three groups (P>0.05). Six months after operation, Lysholm score 80.74±3.14 and IKDC score 79.92±3.44 in group A were higher than those in group B 80.74±3.14 and 78.21±4.63, and higher than those in group C 79.22±3.63 and 76.63±3.80 (P<0.05);12 months after operation, Lysholm score 89.84± 5.13 and IKDC score 87.90±3.93 in group A were higher than those in group B 85.74±6.04 and 83.62±5.64, and higher than those in group C 82.83±3.43 and 79.21±4.04(P<0.05). CONCLUSION: Compared with remnant-non-preserving group, the residual tissue of anterior cruciate ligament is preserved, which is conducive to promote the healing and remodeling of tendon graft and accelerate the recovery of joint function. Proper fixation of residual tissue and restoration of its tension are the key factors affecting the postoperative efficacy.

[Biomechanical study on different high-strength sutures and suture site for repairing posterior root tear of the medial meniscus].

OBJECTIVE: To compare biomechanical characteristic of different high-strength sutures and suture sites for repairing posterior root tear of the medial meniscus with modified Mason-Allen technique. METHODS: Forty-eight specimen of medial meniscus of knee joint from fresh porcine (female, aged from 5 to 9 months with an average of 7 months) were chosen and established experimental model. The samples were divided into red zone fixation group and red-white zone fixation group according to suture sites, 24 in each group; and then were randomly divided into 3 subgroups which 8 in each group, and fixed with Ethibond suture, Ultrabraid suture and FiberWire suture, respectively. Biomechanical tests were performedon universal electromagnetic and mechanical testing machine. Each specimen was underwent 1 000 cyclic tests on the first time, then pull out test until failure. The maximum failure load, yield load, stiffness and displacement were analyzed. RESULTS: All specimen were successfully completed biomechanical tests. The failure mode of Ethibond group was caused by suture fracture; 6 cases of Ultrabraid suture group was caused by suture fracture which belong to red zone fixation group, 10 cases were caused by suture pull out, which 2 cases belong to red zone fixation group, 8 cases belong to red-white zone fixation group;8 cases of FiberWire group was caused by suture pull-out. Biomechanical test showed that:(1)In terms of suture strength, comparison of the maximum failure load, yield load and stiffness showed that Ethibond suture group Ultrabraid suture group >FiberWire suture group, and had statistical differences among groups (P<0.05);the results showed the suture strength in Ethibond was the best, Ultrabraid suture took the second place, and FiberWire suture was the worst. (2)As for different suture sites, comparison of the maximum failure load, yield load and stiffness showed that red zone fixation group >red-white region fixation group, and had statistical difference between two groups(P< 0.05);comparison of cyclic displacement at 100, 500 and 1 000 cycles showed that red zone fixation group

Genome-Wide Identification and Co-Expression Analysis of ARF and IAA Family Genes in Euscaphis konishii: Potential Regulators of Triterpenoids and Anthocyanin Biosynthesis.

Euscaphis konishii is an evergreen plant that is widely planted as an industrial crop in Southern China. It produces red fruits with abundant secondary metabolites, giving E. konishii high medicinal and ornamental value. Auxin signaling mediated by members of the AUXIN RESPONSE FACTOR (ARF) and auxin/indole-3-acetic acid (Aux/IAA) protein families plays important roles during plant growth and development. Aux/IAA and ARF genes have been described in many plants but have not yet been described in E. konishii. In this study, we identified 34 EkIAA and 29 EkARF proteins encoded by the E. konishii genome through database searching using HMMER. We also performed a bioinformatic characterization of EkIAA and EkARF genes, including their phylogenetic relationships, gene structures, chromosomal distribution, and cis-element analysis, as well as conserved motifs in the proteins. Our results suggest that EkIAA and EkARF genes have been relatively conserved over evolutionary history. Furthermore, we conducted expression and co-expression analyses of EkIAA and EkARF genes in leaves, branches, and fruits, which identified a subset of seven EkARF genes as potential regulators of triterpenoids and anthocyanin biosynthesis. RT-qPCR, yeast one-hybrid, and transient expression analyses showed that EkARF5.1 can directly interact with auxin response elements and regulate downstream gene expression. Our results may pave the way to elucidating the function of EkIAA and EkARF gene families in E. konishii, laying a foundation for further research on high-yielding industrial products and E. konishii breeding.

[Arthroscopic repair of Bankart lesion with bioabsorbable suture anchors].

OBJECTIVE: To evaluate early clinical effects of bioabsorbable suture anchors for the treatment of Bankart lesion. METHODS: Total 23 patients with the Bankart lesion were treated with arthroscopic repair using bioabsorbable suture anchors from January 2010 to June 2017. There were 20 males and 3 females, with an average age of (23.4±3.9) years old (ranged, 19 to 34 years old). Fourteen patients had injuries on the right shoulder joint and 9 patients had the injuries on the left side. The mechanism of primary dislocation included 17 cases of training, 5 cases of sports injury and 1 case of falling down. The mean interval time from injury to surgery was(10.9±5.8) months (ranged, 3 to 36 months). The Bankart lesion was repaired by bio-cortical suture anchors. The Rowes rating system for Bankart repair was used to evaluate therapeutic effects. RESULTS: All 23 patients were followed up, with a mean duration of(24.5±3.7) months(ranged, 18 to 39 months). At the latest follow up, there was no recurrent dislocation occurred, and all patients had returned to sports and work. The Rowes rating system for Bankart repair was 53.91±11.67 pre-operationally and 91.74±12.30 post operationally, respectively (P<0.01). According to the Rowes rating system, there were 0 case of excellent, 0 case of fine, 9 cases of good and 14 cases of bad pre-operationally;16 cases of excellent, 4 case of fine, 3 cases of good and 0 cases of bad post operationally;the difference was statistically significant (P<0.01). CONCLUSION: Applying bio-cortical bone suture anchors for the Bankart lesion is a reliable, efficient and cost effective treatment, which is also suitable for the revision of the Bankart lesion.

Tetra-primer ARMS-PCR combined with GoldMag lateral flow assay for genotyping: simultaneous visual detection of both alleles.

Rapid and simple detection of single nucleotide polymorphism (SNP) is vital for individualized diagnosis and eventual treatment in the current clinical setting. In this study, we developed a tetra-primer ARMS-PCR combined lateral flow assay (T-ARMS-PCR-LFA) method for simultaneous visual detection of two alleles. By using four primers labeled with digoxin, biotin and Cy5 separately in one PCR reaction, the amplified allele-specific products could be captured by streptavidin and the anti-Cy5 antibody on two separated test lines of a LFA strip, which allows the presentation of both alleles within the single LFA strip. Both DNA and whole blood can be used as templates in this genotyping method in which the whole detection process is completed within 75 minutes. The performance assay of T-ARMS-PCR-LFA demonstrates the accuracy, specificity and sensitivity of this method. One hundred human whole blood samples were used for MTHFR C677T genotyping in T-ARMS-PCR-LFA. The concordance rate of the results detected was up to 100% when compared with that of the sequencing results. Collectively, this newly developed method is highly applicable for SNP screening in clinical practices.

Direct genotyping from whole blood using alkaline polyethylene glycol.

In order to simplify biological sample preparation to meet the demand of rapid genotyping, we improve alkaline polyethylene glycol (APEG) based on Chomczynski's procedure for the PCR-based lateral flow genotyping system, which enables the rapid and efficient direct genotyping from whole blood without DNA purification. The improved APEG has a high tolerance for extreme storage conditions. Testing whole blood with an abnormal hematological index indicates that APEG efficiency would not be influenced by pathological factors. Compared with sequencing, the accuracy of this genotyping system was 100% on testing with 200 clinical samples.

Genotyping of Multiple Clinical Samples with a Combined Direct PCR and Magnetic Lateral Flow Assay.

Developing a sensitive, low-cost, and easy-to-use point-of-care testing system for genotyping is important for informing treatment decisions and predicting the risk of underlying diseases. Conventional methods normally require complex operational procedures as well as expensive and sophisticated instruments. Here, we report a general approach that enables us to detect the genotype of multiple sample types directly without DNA purification. Moreover, the PCR results can be further quantitatively analyzed based on a magnetic lateral flow assay (MLFA) system, which avoids multiple steps needed for conventional nucleic acid biosensors. As a demonstration, we show that three genotypes of aldehyde dehydrogenase 2 (ALDH2) can be identified using a small volume of sample with an accuracy of 100% and a sensitivity of 1.0 × 102 cells/µL, which are better than those of the gold standard methods. We believe that the direct PCR-MLFA system represents a significant advance toward the development of portable, sensitive biomedical platforms.

A direct isothermal amplification system adapted for rapid SNP genotyping of multifarious sample types.

Genotyping of single nucleotide polymorphisms (SNPs) in point-of-care (POC) settings could be further improved through simplifying the treatment of samples. In this study, we devised an accurate, rapid and easy-to-use SNP detection system based on direct loop-mediated isothermal amplification (LAMP) without DNA extraction, known as Direct-LAMP. Samples from various sources (including whole blood, dried blood spot, buccal swab and saliva), treated with NaOH, can be used directly in amplification. The turnaround time was about 30 min from sample collection to provision of results. The accuracy was evaluated by assessing the polymorphisms of methylenetetrahydrofolate reductase (MTHFR) C677T and aldehyde dehydrogenase-2 (ALDH2) Glu504Lys, which are better known for their critical role in folate and ethanol metabolism, respectively. Completely consistent genotyping results reveal that Direct-LAMP is generally concordant with sequencing. This system can serve as a very promising platform in the fields of disease predisposition, drug metabolism and personalized medicine.

Multiple SNPs Detection Based on Lateral Flow Assay for Phenylketonuria Diagnostic.

Single nucleotide polymorphisms (SNPs) are closely related to genetic diseases, but current SNP detection methods, such as DNA microarrays that include tedious procedures and expensive, sophisticated instruments, are unable to perform rapid SNPs detection in clinical practice, especially for those multiple SNPs related to genetic diseases. In this study, we report a sensitive, low cost, and easy-to-use point-of-care testing (POCT) system formed by combining amplification refractory mutation system (ARMS) polymerase chain reaction with gold magnetic nanoparticles (GMNPs) and lateral flow assay (LFA) noted as the ARMS-LFA system, which allow us to use a uniform condition for multiple SNPs detection simultaneously. The genotyping results can be explained by a magnetic reader automatically or through visual interpretation according to the captured GMNPs probes on the test and control lines of the LFA device. The high sensitivity (the detection limit of 0.04 pg/µL with plasmid) and specificity of this testing system were found through genotyping seven pathogenic SNPs in phenylalanine hydroxylase gene ( PAH, the etiological factor of phenylketonuria). This system can also be applied in DNA quantification with a linear range from 0.02 to 2 pg/µL of plasmid. Furthermore, this ARMS-LFA system was applied to clinical trials for screening the seven pathogenic SNPs in PAH of 23 families including 69 individuals. The concordance rate of the genotyping results detected by the ARMS-LFA system was up to 97.8% compared with the DNA sequencing results. This method is a very promising POCT in the detection of multiple SNPs caused by genetic diseases.

Sensitive genotyping of mutations in the EGFR gene from NSCLC patients using PCR-GoldMag lateral flow device.

Epidermal growth factor receptor (EGFR) mutations predict better outcomes with EGFR tyrosine kinase inhibitors in patients with non-small cell lung cancer (NSCLC). Most common activating mutations include in-frame deletion in exon 19 and L858R substitution in exon 21, which account for >90% of all EGFR mutations in NSCLC. In this study, a PCR-GoldMag lateral flow assay (PCR-GoldMag LFA) was developed for the visual detection of delE746-A750 and L858R of EGFR mutations. Forty formalin-fixed paraffin-embedded (FFPE) tissue samples of NSCLC patients were analyzed using PCR-GoldMag LFA system and verified by direct sequencing and TaqMan-PCR detection methods. Results showed that EGFR mutations were detected in 34 cases among the 40 samples (85%) by PCR-GoldMag LFA method. Among the 34 cases, 5 cases were simultaneously detected with delE746-A750 in exon 19 and L858R mutation in exon 21. Compared with sequencing, only 4 samples were detected as delE746-A750, which revealed higher sensitivity of PCR-GoldMag LFA detection method than direct sequencing. TaqMan-PCR method verified the L858R mutation and was in 100% agreement with our method. These results indicated that our method has obvious advantages to analyze clinical samples and offers a more sensitive alternative to direct sequencing for the detection of EGFR mutations.

[Midterm clinical outcome for ankylosing spondylitis patients with early hip-involved diseases treated with arthroscopic technique].

OBJECTIVE: To study the clinical outcome of manipulation release combined with arthroscopic debridement and synovia resection under general anesthesia for early hip involvment in patients with ankylosing spondylitis. METHODS: Manipulation release combined with arthroscpic debridement and synovium resection were performed for hip lesion in 22 patients with ankylosing spondylitis from June 2011 to June 2013, incuding 6 males and 16 females with anverage age of 24.7 years ranging from 17 to 23 years. The course of the diseases was from 10 to 41 months(22.1 months on average). After 6 months of conservative treatment, hip pain and other symptoms were no relief. The preoperative and postoperative follow-up evaluation was performed and compared by the hip movement, VAS pain score, mHHS score and NAHS score. RESULTS: All the patients were followed up for 26 to 44 months with an average of 30.2 months. The range of motion in active flexion-extension, abduction-adduction, internal-external rotation in 0° flexion and 90° flexion increased from (78.2±10.2)°, (36.3±6.4)°, (31.1±9.2)° and (37.3±10.5)° before operation to (113.5±8.4)°, (55.7±8.4)°, (58.7±2.1)° and (60.1±9.8)° after operation, respectively. The VAS scores decreased from 8.5±9.4 before operation to 5.5±7.1 after operation. The modified Harris and NAHS scores increased from 60.8±6.9 and 56.9±6.25 before operation to 88.1±10.4 and 84.6±5.4 after operation, respectively. CONCLUSIONS: Manipulation release combined with arthroscopic debridement and synovium resection under general anesthesia could effectively control the progression of hip lesion in patients with ankylosing spondylitis restoring the ROM, relieve pain symptoms, delay joint deformity and ankylosis with less bleeding, faster recovery, and significantly improve patients' quality of life.

[Experimental study on staphylococcal enterotoxin promoting tendon-bone healing after reconstruction of anterior cruciate ligament in rabbits].

OBJECTIVE: To explore highly agglutinative staphylococcin (HAS) pomoting on tendon-bone healing after reconstruction of anterior cruciate ligament(ACL) in rabbits. METHODS: Animal model of ACL reconstruction in 24 mature New Zealand white rabbits(3 months, 2.56 kg on average, either gender) were established using autologous digital long extensor tendon and randomly classified into 2 groups(HAS and control group), 12 rabbits for each group. HAS group was separately injected 0.1ml highly agglutinative staphylococcin immediately into tendon-bone interface during the operation and 2 days after operation. Control group was injected with the same dose of physiological saline for 3 days. All animals were sacrificed at 4, 8, and 12 weeks after operation for histological examinations. The specimens were stained with hematoxylin-eosin, picric acid-sirius red, VEGF immunohistochemistry stain, and toluidine blue to histologically analysis the total pathology changes of the tendon-bone healing tissue, the tendon bone interface morphology classification, hyperplasia and arrangement of collagen fiber, vascularization and new bone formation, respectively. RESULTS: The Yamakado morphological interface results showed that the tissue healing at tendon-bone interface of the HAS group was better than that of the control group. The histological examination revealed that on the 4th week after operation, the tendon-bone interface of HAS group was filled with fibrous connective tissue. The proliferated fibroblasts, chondroblasts and the angiogenesis were rich. On the 8th week after operation, the healing tissue at the bone-tendon interface had developed into dense connective tissue, the neo-vessels were very rich, the collagen fibers were formed abundantly, some Sharpey's fibers spanning parts of the tendon-bone interface. On the 12th week after operation, the transition zones were full of Sharpey's fibers;the neo-vessels were not as much as the 8th weeks, but new bone formation was further increased and immature fibrocartilage appeared. For quantitative histological analysis at 4, 8 and 12 weeks after operation, the proportion of neo-vessel area and the area of now bone formation of the HAS group were all significantly higher than those of the control group(P<0.05). CONCLUSIONS: Under the synergy of staphylococcal enterotoxin C and other active ingredients, Highly Agglutinative Staphylococcin can significantly improve the tendon-bone healing after ACL reconstruction in rabbit knees, which is expected to be a new method to improve the clinical results of ACL reconstruction.

[Clinical outcome of arthroscopic anterior cruciate ligament remnant-preserving reconstruction].

OBJECTIVE: To evaluate the clinical outcome of arthroscopic anterior cruciate ligament(ACL)reconstruction with remnant cuff preservation. METHODS: There were 42 cases of ACL tear were performed reconstruction in our department from January 2012 to December 2014. The patients were 28.4 years old on average, 17 males and 25 females. ACL reconstruction was performed under arthroscopy with remnant cuff preservation. The stability of knee was assessed by Lachman test and anterior drawer test. The function of knee was assessed through Lysholm score and Tegner activity rating. MRI of the knee was checked 12 months postoperatively. RESULTS: The stability tests of all patients were negative postoperatively. Lysholm score of all patients 12 months after operation was 96.8±6.1, which was significantly better than 37.8±7.1 of preoperatively. Tegner activity rating of all patients 12 months after operation was 6.2±0.9, which was significantly better than 2.1±0.4 of preoperatively. It showed the grafts were very well in MRI 12 months after operation. CONCLUSIONS: Arthroscopic anterior cruciate ligament remnant-preserving reconstruction could restore the stability of knee.

Apolipoprotein E genotyping using PCR-GoldMag lateral flow assay and its clinical applications.

A polymerase chain reaction-gold magnetic nanoparticles lateral flow assay (PCR-GoldMag LFA) has been developed via integrating multiplex amplification refractory mutation system PCR (multi­ARMS­PCR) with GoldMag­based LFA for the visual detection of single­nucleotide polymorphisms (SNPs). This assay was applied to genotype Apolipoprotein E (ApoE). ApoE genotyping is important due to the predictive value for the development of coronary artery disease and Alzheimer's disease. The method requires two steps: i) Simultaneous amplifications of the two polymorphic codons (ApoE 158 and 112), performed in separated reactions using multi­ARMS­PCR; and ii) detection of the wild­type and mutant PCR products via dual immunoreactions, which can be performed in ~5 min. Within two LFAs, anti­digoxin antibody­conjugated GoldMag probes bind digoxin­labeled wild­type PCR products, and anti­fluorescein isothiocyanate (FITC) antibody-conjugated GoldMag probes bind FITC­labeled mutant PCR products. All PCR products are biotin labeled and are detected by streptavidin-coated regions on the LFA strip, resulting in a red color. The current approach is capable of detecting the SNPs of ApoE in ~1.5 h, with a broad detection range from 10­1,000 ng of genomic DNA. Thus, the present protocol may facilitate simple, fast and cost­effective screening for important SNPs, as demonstrated by the evaluation of the prevalence of ApoE variants in a Han Chinese cohort.

Establishment and application of a real-time loop-mediated isothermal amplification system for the detection of CYP2C19 polymorphisms.

Single-nucleotide polymorphisms (SNPs) represent the most widespread type of genetic variation (approximately 90%) in the human genome, and the demand to overcome such variation has received more attention now than ever before. The capacity to rapidly assess SNPs that correlate with disease predisposition, drug efficacy and drug toxicity is a key step for the development of personalized medicine. In this work, a rapid one-step SNP detection method, real-time loop-mediated isothermal amplification (RT-LAMP), was first applied for CYP2C19 polymorphisms testing. The optimized method was established with specifically designed primers for target amplification by real-time detection in approximately 30 min under isothermal conditions. RT-LAMP amplified few copies of template to produce significant amounts of product and quantitatively detected human DNA with compatible specificity and sensitivity. The success in the establishment of this RT-LAMP protocol for CYP2C19 polymorphism testing is significant for the extension of this technique for the detection of other SNPs, which will further facilitate the development of personalized medicine.

[Clinical effect of arthroscopy-assisted minimally invasive management of bunion].

OBJECTIVE: To evaluate the effect of arthroscopy-assisted minimally invasive management of bunion and hallux valgus deformities. METHODS: Total 50 patients (53 feet) with bunion and hallux valgus deformities were treated under arthroscopy from July 2008 to July 2011, with an average age of 42.3 years old (ranging from 30 to 65 years old) involving 19 left feet, 28 right feet and 3 both feet. The American Orthopedic Foot and Ankle Society (AOFAS) hallux metatarsophalangeal-interphalangeal(MP-IP) Scale Score was used to evaluate the therapeutic effect. RESULTS: There were no complications such as hallux varus, hallux rigid and nerve or blood vessel injury. Clinically, AOFAS MP-IP Scale Score was increased from 62.19 ± 6.01 preoperatively to 88.26 ± 6.81 postoperatively. CONCLUSION: Arthroscopy-assisted minimally invasive management appears to be a good procedure with advantages of less complication, little trauma and early rehabilitation for bunion and hallux valugs deformities.

A novel lateral flow assay based on GoldMag nanoparticles and its clinical applications for genotyping of MTHFR C677T polymorphisms.

Current techniques for single nucleotide polymorphism (SNP) detection require tedious experimental procedures and expensive and sophisticated instruments. In this study, a visual genotyping method has been successfully established via combining ARMS-PCR with gold magnetic nanoparticle (GoldMag)-based lateral flow assay (LFA) and applied to the genotyping of methylenetetrahydrofolate reductase (MTHFR) C677T. C677T substitution of the gene MTHFR leads to an increased risk of diseases. The genotyping result is easily achievable by visual observation within 5 minutes after loading of the PCR products onto the LFA device. The system is able to accurately assess a broad detection range of initial starting genomic DNA amounts from 5 ng to 1200 ng per test sample. The limit of detection reaches 5 ng. Furthermore, our PCR-LFA system was applied to clinical trials for screening 1721 individuals for the C677T genotypes. The concordance rate of the genotyping results detected by PCR-LFA was up to 99.6% when compared with the sequencing results. Collectively, our PCR-LFA has been proven to be rapid, accurate, sensitive, and inexpensive. This new method is highly applicable for C677T SNP screening in laboratories and clinical practices. More promisingly, it could also be extended to the detection of SNPs of other genes.

[The value of 3 dimensional-fat suppression-spoiled gradient-recalled acquisition sequence on single compartment osteoarthritis for unicompartmental arthroplasty preoperative assessment].

OBJECTIVE: To analyze the 3 dimensional-fat suppression-spoiled gradient-recalled acquisition (3D-FS-SPGR) sequence in the diagnosis of knee articular cartilage injury. METHODS: A total of 56 knee osteoarthritis patients (26 males, 30 females, ages 52-73 years, mean 61.8 years) treated in Department of Orthopedics, Chinese People's Liberation Army General Hospital between June 2013 and May 2014 were involved in this study. All patients underwent knee MRI, plus 3D-FS-SPGR sequence, arthroscopic exploration, and in contrast to the results of MRI results analysis, evaluation 3D-FS-SPGR and conventional sequence of cartilage damage consistent with the arthroscopic accuracy. RESULTS: Divided 56 knee joints into 336 cartilage articular surface, included 55.1% normal articular surface, 21.4% early osteoarthritis and 23.5% advanced osteoarthritis. The accordance of 3D-FS-SPGR sequence grading and arthroscopic was 90.2%. The sensitivity of 3D-FS-SPGR sequence was 93.1%, specificity was 98.3%, and Kappa value was 0.849. The sensitivity of T2WI sequence was 84.4%, specificity was 96.9%, and the Kappa value was 0.671. CONCLUSION: For unicompartment osteoarthritis , MRI 3D-FS-SPGR sequence is effective in sensitivity and specificity of cartilage damage.

Determination of ABO blood group genotypes using the real­time loop­mediated isothermal amplification method.

ABO genotyping is commonly used in several situations, including blood transfusion, personal identification and disease detection. The present study developed a novel method for ABO genotyping, using loop­mediated isothermal amplification (LAMP). This method allows the simultaneous determination of six ABO genotypes under 40 min at a constant temperature of 62˚C. The genotypes of 101 blood samples were determined to be AA (n=6), AO (n=38), BB (n=12), BO (n=29), AB (n=8) and OO (n=8) by the LAMP assay. The results were compared with the phenotypes determined by serological assay and the genotypes determined by direct sequencing, and no discrepancies were observed. This novel and rapid method, with good accuracy and reasonably cost effective, provides a supplement to routine serological ABO typing and may also be useful in other point­of­care testing.