OBJECTIVE Based on fibroblast activation protein(FAP)gene promoter as the response element,to develop a new dual luciferase reporting system for the screening of drugs related to myocardial fibrosis.METHODS The promoter fragment of mouse FAP gene was synthesized in vitro and cloned into plasmid psiCHECK2 to replace HSV-TK promoter,and then a new recombinant plasmid psiCHECK2-FAP was obtained.After the recombinant plasmid psiCHECK2-FAP was digested by restriction endonucliase Hind Ⅲ,the product digested was identified by agar-gel electrophoresis and sequencing.After psiCHECK2-FAP was transient transfected into mouse cardiac fibroblasts(MCFs),and continued cultured for 24 h,and MCFs were treated with Ransforming growth factor β1(TGF-β1,5 μg·L-1)or angiotensinⅡ(Ang Ⅱ,1 μmol·L-1)or palmitic acid(PA,100 μmol·L-1)for 0,12,24,48 h,respectively,the double luciferase reporter gene assay was used to detect luciferase activity;After psiCHECK2-FAP was transient transfected into MCFs,the cells were pretreated with Dapa(1 μmol·L-1)for 1 h,and supplemented with TGF-β1(5 μg·L-1)or AngⅡ(1 μmol·L-1)or PA(100 μmol·L-1),continued treatment for 24 h,the double luciferase reporter gene assay was used to detect luciferase activity,and the expression of collagenⅠand collagen Ⅲ protein was detected with Western blotting.RESULTS The recombinant plasmid psiCHECK2-FAP was digested into two fragments by Hind Ⅲ with the expected strip size,and the sequencing results were completely consistent with the theoretical sequence;Compared with control group,the collagenⅠand collagen Ⅲ protein expression were significantly increased by TGF-β1 or Ang Ⅱ or PA in MCFs(P<0.05,P<0.01).However,compared with TGF-β1 or Ang Ⅱ or PA group,the intervention of Dapa significantly alleviated the promoter activity of FAP gene and the expression of collagenⅠand collagen Ⅲ protein(P<0.05,P<0.01);Compared with control group,luciferase activity was significantly increased by TGF-β1 or Ang Ⅱ or PA(P<0.05,P<0.01),reaching the peak at 24 h.Compared with TGF-β1 or AngⅡ or PA group,the intervention of Dapa significantly decreased luciferase activity(P<0.05,P<0.01).CONCLUSSION Based on the promoter of FAP gene as the response element,a noval dual luciferase reporter gene system was established and showed a good sensitivity to the promyocardial fibrosis factor in MCFs,which can provide strategies for the development of antimyocar-dial fibrosis drugs.

Objective:To evaluate the clinical efficacy of modified all-arthroscopic reconstruction of medial patella femoral ligament (MPFL) for the treatment of recurrent patellar dislocation.Methods:A retrospective case series study was conducted to analyze the clinical data of 38 patients (46 knees) with recurrent patellar dislocation, who were treated at First Affiliated Hospital of Shenzhen University from January 2017 to January 2020. The patients included 12 males (12 knees) and 26 females (34 knees), aged 14-40 years [(24.6±5.4)years]. All patients underwent the modified all-arthroscopic MPFL reconstruction procedure. The femoral tunnel locations were assessed by 3D-CT immediately after surgery. The MRI was performed at 6 and 12 months after operation to assess the healing morphology of the reconstructed MPFL. The Lysholm score and Kujala score were used to assess the knee function before operation, at 6 months after operation, at 12 months after operation and at the last follow-up. The time to return to sports as well as complications were observed.Results:All patients were followed up for 26-48 months [(32.4±8.6)months]. Postoperative 3D-CT examination showed that the femoral tunnels were located in the groove area of the medial epicondyle of the femur and the adductor tubercle. At 6 and 12 months after operation, MRI T2 images showed that the reconstructed MPFL had a low signal and well tensioned ligament tissue, indicating that the MPFL was healed well. The Lysholm scores at 6 and 12 months postoperatively and at the last follow-up were (81.1±12.0)points, (91.2±3.8)points, and (92.2±9.8)points, respectively, being significantly higher than the preoperative (52.4±10.6)points (all P<0.01). The Kujala scores at 6 and 12 months postoperatively and at the last follow-up were (85.4±3.9)points, (91.4±3.6)points, and (93.1±8.5)points, respectively, being significantly higher than the preoperative (55.2±6.8)points (all P<0.01). Compared with 6 months postoperatively, the Lysholm score and Kujala score were significantly improved at 12 months postoperatively and at the last follow-up (all P<0.05). All patients returned to sports, with the time to return to sports for 3-12 months [(8.7±2.3)months] after operation. One patient had poor wound healing but was healed after dressing changes. No wound infection, nerve injury, joint stiffness, patella re-dislocation or other complications occurred. Conclusion:For recurrent patellar dislocation, the modified all-arthroscopic MPFL reconstruction has advantages of accurate bone tunnel positioning, good ligament healing, good function recovery, early return to sports, and less postoperative complications.

OBJECTIVE: To compare the clinical outcomes and complications of hip arthroscopic treatment for femoroacetabular impingement (FAI) performed with either Inside-out or Outside-in approach. METHODS: The clinical date of 48 patients with FAI treated by hip arthroscopy surgery and follow-up from June 2016 to June 2019 were retrospectively analyzed. According to the different operative methods, the patients were divided into two groups. Inside-out group, from central compartment to peripheral compartment;Outside-in group, from peripheral compartment to central compartment. There were 14 males and 10 females in Inside-out group with an averageage of (39.8±7.6)years old, 13 males and 11 females in Inside-out group with an average age of (39.5±9.1)years old in Outside-in group. There was no significant difference in age, gender, body mass index, side, impingement type, medical history and follow-up time between the two groups. The complication occurrence rate, modified Harris hip score (mHHS)and nonarthritic hip score (NAHS) were compared between these two groups. RESULTS: The mHHs and NAHS scores of the two groups were significantly higher than those before operation, but there was no significant difference between the two groups ( CONCLUSION Both hip arthroscopic surgery methods can obtain satisfactory clinical efficacy in the treatment of FAI, but the incidence of postoperative complications of Outside-in surgical method is lower. The out-side in method can be preferentially selected for the patients with the indications of operation.
Adult ; Arthroscopy ; Female ; Femoracetabular Impingement/surgery* ; Follow-Up Studies ; Hip Joint/surgery* ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Gout is a common metabolic disease. Gouty arthritis is associated with the deposition of urate crystals in the synovial fluid or para-articular soft tissues, which often affects the hands, feet, wrists, ankle and knees joints. However, the shoulder joint is unusual involved. In the present study, a 49-year-old male patient complained of right shoulder pain for 9 months and aggravating with limited range of motion for 2 weeks. Physical examination showed that the external rotation was 10° and hand back was S 5 of the right shoulder. The MRI showed shoulder joint effusion and intra-articular loose bodies. The patient was admitted to the hospital with the diagnosis of "loose bodies of the right shoulder with synovial chondromatosis possibly". Arthroscopic examination revealed that a lots of urate crystals were deposited on synovium, cartilage and rotator cuff. Loose bodies removal and joint debridement were performed. Histopathologic examination of the specimen demonstrated synovitis hyperplasia with urate crystals deposition. Shoulder gouty arthritis characterized by pain and limited movement. The imaging of tophi depends on the calcium-containing deposits. Tophus with calcium deposits can be found on X-ray and CT. MRI can not only evaluate the tophus but also the evaluate the other intra-articular pathologies. Due to no typical manifestations and specific images of shoulder gouty arthritis, pathological diagnosis is the "gold standard". Shoulder arthroscopy provide a minimal invasive technique in diagnosing and treating of the shoulder disease. Undoubtedly, it could be a reliable diagnosis and treatment method for shoulder gouty arthritis.

No abstract available.
Arthroscopy ; Hip

Objective To investigate the differences in expression of Mohawk (MKX) transcription factors and collagen of types Ⅰ and Ⅲ in anterior cruciate ligament (ACL) gratis between 2 remodeling outcomes under arthroscopy.Methods Enrolled for this study were 17 patients who had undergone arthroscopic single-bundle ACL reconstruction with autogenous hamstring tendons and secondary arthroscopic exploration 48 to 131 months (average,83.1 months) after removal of tibial internal fixator at Department of Sports Medicine,The First Affiliated Hospital to Shenzhen University from March 2017 to December 2017.They were divided into a good remodeling group (11 cases) and a fair remodeling group (6 cases) according to the graft quality under arthroscopy (synovial and vascular coverage,and apparent tension,thickness and retear of the grafts).During the secondary arthroscopic procedures,biopsy of the central ACL grafts was performed.Moreover,normal ACL tissues were harvested from 8 contemporary controls of ＜ 60 years old who underwent total knee replacement.Immunohistochemical assay and quantificational real-time polymerase chain reaction were conducted to detect the expression of transcription factors and collagen of types Ⅰ and Ⅲ in all the samples.Results In the samples from good remodeling and control groups,there were abundant well-arranged collagen fibers of types Ⅰ and Ⅲ and MKX-positive cells;in the fair remolding group,the collagen fibers of types Ⅰ and Ⅲ and MKX-positive cells were much decreased in number and the fibers were not well arranged.The former 2 groups scored in immunohistochemical assay significantly higher than the latter one (P ＜ 0.05).qRT-PCR showed that the expression levels of MKX gene (0.44 ± 0.30),COL1A1 gene (0.52 ± 0.27) and COL3A1 gene (0.60 ± 0.22) in the fair remolding group were significantly lower than in the control group (1.00 ± 0.00,1.00 ± 0.00 and 1.00 ± 0.00) and than in the good remolding group (0.97 ± 0.67,0.99 ±0.38 and 1.00 ± 0.35) (P ＜ 0.05).Conclusion Good remodeling ACL grafts with histological maturation under arthroscopy are more similar to normal ACL than fair remodeling ACL grafts in expression of MKX transcription factors and collagen of types Ⅰ and Ⅲ.

BACKGROUND:The use of normal hyaline cartilage to repair large areas of full-thickness knee cartilage defect has been a hot topic recently; however, a follow-up study with a relative large number of patients is required. OBJECTIVE:To make a preliminary study concerning the methods and therapeutic effects of tissue-engineered cartilage (TEC) implantation for treating large-area full-thickness knee cartilage defects. METHODS:Twenty-one patients (23 knees) diagnosed with cartilage defect of the knee joint (Outbridge III-IV) were enrolled. The area of the cartilage defect was 3.5-11.2 cm2. All of the patients were given TEC treatment. Postoperative functional exercise of the knee joint was carried out in these patients as planned. We regularly reviewed the knee MRI and calculated visual analog scale score, International Knee Documentation Committee (IKDC) score, and Lysholm score. RESULTS AND CONCLUSION:All the patients were followed up for 3 to 12 months. Postoperatively knee pain relieved obviously, and the visual analog scale score was significantly declined compared with the preoperation (P<0.05). All the patients manifested painless 1 year after surgery. The 1-year postoperative MRI showed that the injured cartilage grew well. The thickness and MRI signal of the graft was the same as the normal cartilage, and the bone healed completely. The IKDC and Lysholm scores were significantly improved at 3, 6, 12 months after the surgery, and the difference was statistically significant before and after the surgery (P<0.05). Overall, TEC is an improved technique of chondrocyte implantation, which is an effective and safe method for cartilage defect repair.

Objective To observe the therapeutic efficacy and safety of recombinant tissue plasminogen activator (rt-PA) for treatment of patients with acute cerebral infarction and investigate the prognostic factors of thrombolysis. Methods The clinical data of 60 patients with acute cerebral infarction from January 2009 to November 2013 in Department of Emergency, Hangzhou Traditional Chinese Medicine Hospital were retrospectively analyzed;of them, 30 cases received intravenous rt-PA thrombolytic treatment, being in the thrombolytic group, 0.9 mg/kg rt-PA was given to the patient within 4.5 hours after the disease onset, the total dosage could not exceed 90 mg, in which 10%was intravenously injected and the rest 90%was intravenously dripped slowly within 60 minutes. That another 30 cases did not undergo thrombolytic therapy was assigned as the control group, and they took aspirin, etc anti-platelet aggregation routine treatment. Before and after thrombolytic treatment for 1 hour, 24 hours and 14 days, the National Institutes of Health Stroke Scale (NIHSS) score was evaluated in the two groups;before and after thrombolytic therapy for 0.5, 1.0, and 1.5 hours, the patient's scores of Barthel index (BI) were observed in the two groups. In thrombolytic group, the situations of hemorrhage transformation, symptomatic hemorrhage and modified Rankin scale (mRS) score in 3 months of the patients with different ages, complications and NIHSS scores were observed. Results Before treatment, no statistically significant differences were found in the NIHSS score (15.2±3.6 vs. 15.5±3.3) and BI score (45.0±8.8 vs. 44.1±7.6) between the control group and thrombolytic group (both P>0.05);after treatment with the extension of time, the NIHSS score was gradually reduced, reaching the lowest level on the 14th day after thrombolytic treatment, while the BI score was gradually increased, reaching its peak at 1.5 hours after thrombolytic treatment, the changes being more prominent in thrombolytic group (NIHSS score:9.7±2.6 vs. 12.8±4.2, BI score:82.6±7.8 vs. 69.6±9.8, both P<0.05). In thrombolytic group, there were cerebral hemorrhage transformation 2 cases, gum bleeding 3 cases, skin bleeding 1 case, urethral bleeding 1 case, gastrointestinal bleeding and black stool 1 case. In control group, cerebral hemorrhage transformation was seen in 1 case. There was no obvious systemic hemorrhage in the two groups. In the 30 cases in thrombolytic group, the baseline NIHSS score of patients>80 years old was higher than that in cases≤80 years old (15 vs. 12); the age (years: 71.0±4.1 vs. 61.5±2.6), baseline NIHSS score (14 vs. 11) and bleeding conversion rate [37.50% (3/8) vs. 18.18% (4/22)] of cases with atrial fibrillation were higher than those not complicated with atrial fibrillation; the NIHSS score of cases with elevated international normalized ratio (INR) was lower than those without elevated INR (11 vs. 14);The bleeding conversion rate [16.67%(1/6) vs. 29.17%(7/24)] with NIHSS score≤4 were lower than those with NIHSS score>4;the mRS score in 3 months (4 vs. 2), and percentage of 3-month mRS score≤2 [11.1%(1/9) vs. 52.38%(11/21)] in cases with NIHSS score≥20 was higher than that in cases with NIHSS score<20 (all P<0.05). The fatality rate of two groups was 3.33%. Conclusions The intravenous rt-PA thrombolytic treatment can significantly promote the early recovery of neurologic impairment for patients with acute cerebral infarction. The therapy can improve the prognosis and its safety is relatively good. Meanwhile it is similarly effective for cases over 80, with complications such as atrial fibrillation, raised INR, and with different degrees of severity.

Objective To observe the effect of Shenfu injection on intestinal function in rats with sepsis. Methods Forty Sprague-Dawley (SD) rats were randomly divided into four groups: sham operation, sepsis model, low and high concentration Shenfu injection groups, each groupn = 10. The sepsis model was replicated by cecal ligation and puncture (CLP), while the rate in sham operation group just underwent abdominal incision without CLP. Ten minutes after CLP, the low and high dose Shenfu injection groups were given 5 mL/kg and 10 mL/kg Shenfu intravenous injection via a tail vein respectively. The rats in the model group were treated by intravenous injection of 10 mL/kg normal saline through a tail vein in 10 minutes after CLP. Twelve hours later, the rats were sacrificed. The levels of Ghrelin, Gastrin, tumor necrosis factor-α (TNF-α), high mobility group B1 protein (HMGB1), myeloperoxidase (MPO) and diamine oxidase (DAO) activity in serum were detected by enzyme linked immunosorbent assay (ELISA). The levels of protein of Ghrelin and gastrin receptor (GHSR) were detected by Western Blot. Under light microscope, the histopathological changes in intestinal mucosa were investigated, and Chiu score was determined, and the apoptosis index (AI) of intestinal mucosal epithelial cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Results Compared with sham operation group, in model group, the levels of Ghrelin and Gastrin in serum were significantly decreased [Ghrelin (ng/L): 121.23±3.53 vs. 146.28±5.43, Gastrin (ng/L): 81.78±3.27 vs 102.78±4.07], the serum levels of TNF-α and HMGB1 were markedly increased [TNF-α (mg/L): 93.71±3.66 vs. 11.69±1.44, HMGB1 (mg/L): 76.25±4.12 vs. 22.41±3.08], the DAO activity and protein expressions of Ghrelin and GHSR of intestinal tissue were obviously decreased [DAO (U/mL): 14.64 ±0.68 vs. 25.13±1.98, Ghrelin (grey value): 0.12±0.02 vs. 0.23±0.04, GHSR (grey value): 0.18±0.02 vs. 0.32±0.03], the MPO activity in intestinal tissue, Chiu score of intestinal mucosa and AI of ileum mucosal epithelial cells were remarkably increased [MPO (mg/L): 175.98±6.95 vs. 45.64±4.48, Chiu score: 3.90±0.52 vs. 0.30±0.30, AI: 29.31±1.65 vs. 5.45±1.35, allP < 0.01]. Compared with model group, in low and high Shenfu injection groups, the levels of Ghrelin in serum and protein expressions of Ghrelin and GHSR in intestinal tissues were significantly increased (P < 0.05 orP < 0.01), the activity of DAO of intestinal tissues, the Chiu score and AI were significantly decreased, the degrees of changes being more significant in high Shenfu injection group than those in low Shenfu injection group [Ghrelin (ng/L): 143.54±3.89 vs. 136.58±4.91, TNF-α (mg/L): 75.13±4.69 vs. 83.70±4.40, HMGB1 (mg/L): 57.47±4.53 vs. 65.41±4.63, protein expression of Ghrelin (grey value): 0.18±0.03 vs. 0.15±0.03, protein expression of GHSR (grey value): 0.28±0.03 vs. 0.23±0.03, MPO (mg/L): 154.05±5.75 vs. 162.64 ±5.73, DAO (kU/L): 19.70±1.51 vs. 16.67±0.92, Chiu score: 2.30±0.52 vs. 3.20±0.48, AI: 20.38±1.34 vs. 26.40±1.32, allP < 0.05]. The levels of serum Gastrin in low and high Shenfu injection group were higher than those in model group, but no statistically significant differences were found (83.59±3.24, 86.54±5.93 vs. 81.78±3.27, bothP > 0.05). Under light microscope, the pathological changes were seen as follows: destruction and obvious edema of intestinal mucosal villi, ulcer formation, significant perivascular hemorrhage, presence of neutrophil infiltration and fracture of basement membrane in model group, while in low and high Shenfu groups, the intestinal villi had little defect, focal necrosis, small amounts of hemorrhage and neutrophil infiltration. Conclusions Shenfu injection can significantly improve the abnormal expressions of serum Ghrelin, reduce the levels of serum TNF-α and HMGB1, lowered MPO activity and enhance DAO activity in intestinal tissue, alleviate pathological changes in ileum mucosa, and decrease AI of ileum mucosal epithelial cells in rats with sepsis. And the degree of therapeutic effect is proportional to the Shenfu injection dose.