1.Simultaneous Determination of Olanzapine, Risperidone and Paliperidone in Human Plasma by UPLC-MS/MS
Lichun FENG ; Bohan YANG ; Xueqin WANG ; Qiang GUO
China Pharmacy 2017;28(8):1045-1048
OBJECTIVE:To develop a method for the concentration determination of olanzapine,risperidone and paliperidone in human plasma.METHODS:After liquid-liquid extraction,using buspirone hydrochloride as internal standard,the concentration of plasma sample was determined by UPLC-MS/MS.The determination was performed on ACQUITY UPLCTM BEH C18 column with mobile phase consisted of methanol-0.01 mol/L ammonium formate solution (gradient elution) at flow rate of 0.2 mL/min.The column temperature was 45 ℃,and sample size was 5 μL.The electrospray ionization source was adopted for positive ion scanning under MRM mode.Ion-pairs for quantitative analysis were as follows:m/z 313.29→256.25 (olanzapine),m/z 411.42→191.19 (ris peridone),m/z 427.45→207.18 (paliperidone) and m/z 386.43→122.37 (internal standard).RESULTS:The linear ranges of olanzapine,risperidone and paliperidone were 0.426-108.954,0.213-54.476,0.213-54.476 ng/mL,respectively.RSDs of inter-day and intra-day were all lower than 20%.The recoveries of them ranged 83.3%-112.9%,90.0%-109.8% and 95.2%-114.9%,respective ly.Extraction recoveries ranged 65.5%-95.0%,73.9%-98.5% and 73.6%-99.4%,respectively.Both plasma matrix effect and dilute effect didn't influence the determination of plasma concentration.The plasma concentrations of olanzapine,risperidone and paliperidone in 100 schizophrenia patients were (103.3 ± 73.6),(13.1 ± 13.1) and (23.2 ± 20.0) ng/mL,respectively.CONCLU SIONS:The method is simple,rapid,sensitive and specific.It can be used for the determination of plasma concentration and pharmacodynamic study of olanzapine,risperidone and paliperidone.
2.Echocardiographic Evaluation of Left Atrial Structure and Function in Patients with Paroxysmal Atrial Fibrillation After Radiofrequency Catheter Ablation
Sulei LI ; Guang ZHI ; Yang MU ; Bohan LIU
Chinese Journal of Medical Imaging 2017;25(5):370-373
Purpose To observe the changes of left atrial (LA) structure and function in patients with paroxysmal atrial fibrillation after radiofrequency catheter ablation by echocardiography in order to provide basis for clinical evaluation of surgery.Materials and Methods Forty-four patients with paroxysmal atrial fibrillation and treated with radiofrequency catheter ablation in PLA General Hospital from January 2015 to June 2016 were enrolled.According to whether or not to restore sinus rhythm after operation,the patients were divided into sinus rhythm group and atrial fibrillation recurrence group.The paramrters of LA including diameter,maximum and minimum volume,systolic volume,ejection fraction,active ejection fraction,conduit function index and dilatation index were measure by echocardiography before and at least 6 months after radiofrequency catheter ablation.The data were compared between and within groups.Results All patients were followed up for (6.0±0.5) months after ablation operation.29 of 44 patients (66%) maintained sinus rhythm;the anteroposterior,vertical,and left to right diameters of LA in patients with sinus rhythm after operation were significantly lower than those before operation,but the ejection fraction of LA increased (all P<0.05).However,in patients with atrial fibrillation recurrence after operation,the volume of LA increased (P<0.05);the diameters of LA did not show significant differences;the ejection and active ejection fraction of LA had significantly decreased (P<0.05).Compared with patients with sinus rhythm after operation,patients with atrial fibrillation recurrence after operation were older and had higher proportion of hypertension (P<0.05).Conclusion After ablation,the diameter of LA decreases and the ejection fraction increases in patients with sinus rhythm;the volume of LA increases and the function reduces in patients with atrial fibrillation recurrence.
3.Effect of MHSP65-TCL anti-melanoma vaccine on the activity of immunocytes.
Bohan DONG ; Guangli DAI ; Zhilin QI ; Guang YANG ; Jun LU ; Yao ZHANG
Journal of Southern Medical University 2013;33(11):1673-1677
OBJECTIVETo evaluate the therapeutic effect of MHSP65-TCL on melanoma and its effect on the activity of the immunocytes.
METHODSMHSP65-TCL was prepared by mixing MHSP65 with TCL derived from B16 melanoma cell lysate by repeated freezing and thawing. The MHSP65-TCL vaccine was administered in mice bearing B16 melanoma, and the changes in melanoma growth was observed. To investigate the influence of TCL in MHSP65-TCL on the activity of the immunocytes, we co-cultured TCL and mouse spleen cells in vitro, and analyzed CD69 expression on the cells, cell apoptosis, and levels of IL-10 and IFN-γ in the cell culture supernatant.
RESULTSThe MHSP65-TCL vaccine showed an anti-melanoma effect in the tumor-bearing mice. In the in vitro experiment, TCL in MHSP65-TCL strongly stimulated the activation of mouse spleen cells while causing apoptosis in some of the immunocytes and promoting cellular IL-10 secretion, but not IFN-γ.
CONCLUSIONSMHSP65-TCL derived from B16 melanoma cells has an anti-melanoma effect mediated by the activation of immunocytes. TCL in MHSP65-TCL also has immunosuppressive effect on immunocytes possibly due to the presence of suppressive components in TCL, and identifying and eliminating these components may potentially improve the anti-tumor actovoty of MSHP65-TCL vaccine.
Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; Apoptosis ; Bacterial Proteins ; administration & dosage ; immunology ; Cancer Vaccines ; Cell Extracts ; administration & dosage ; immunology ; Cell Line, Tumor ; Chaperonin 60 ; administration & dosage ; immunology ; Female ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Lectins, C-Type ; metabolism ; Melanoma, Experimental ; immunology ; pathology ; Mice ; Mice, Inbred C57BL ; Random Allocation ; Spleen ; cytology ; immunology ; metabolism ; Tumor Burden ; immunology
4.Research progress on Parkinson′s disease with rapid eye movement sleep behavior disorder
Mu YANG ; Yinlian HAN ; Bohan ZHANG ; Yiming LIU
Chinese Journal of Neurology 2024;57(4):393-400
Parkinson′s disease is a common clinical degenerative disease of the nervous system, and rapid eye movement sleep behavior disorder (RBD) is a common sleep symptom of patients with Parkinson′s disease. This article reviews the pathogenesis, epidemiology, clinical characteristics, imaging manifestations, clinical evaluation and treatment of RBD in patients with Parkinson′s disease, in order to deepen the understanding of RBD in patients with Parkinson′s disease.
5.Application value of dual-graft living donor liver transplantation of right segment from an adult living donor combined with a left lateral segment from donation after brain death for hepatocellular carcinoma
Bohan ZHANG ; Jiulin SONG ; Li JIANG ; Jian YANG ; Tao LYU ; Bin HUANG ; Hong WU ; Jiayin YANG ; Lyunan YAN
Chinese Journal of Digestive Surgery 2020;19(2):196-203
Objective:To investigate the application value of dual-graft living donor liver transplantation of right segment from an adult living donor combined with a left lateral segment from donation after brain death for hepatocellular carcinoma (HCC).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of a male 46-year-old patient with HCC who underwent dual-graft living donor liver transplantation of right segment from an adult living donor combined with a left lateral segment from donation after brain death at the West China Hospital of Sichuan University in October 2019 were collected. He weighed 66 kg and was 171 cm in height. His blood type was A Rh-positive. Graft one was from a female 23-year-old living donor who had a bodyweight of 50 kg, a height of 150 cm, and blood type of A Rh-positive; graft two was from a male 44-year-old brain death donor with the blood type of A Rh-positive. The surgery was performed in three operating rooms, graft one and graft two were obtained simultaneously in two operating rooms, and the recipient′s liver was dissected in the third operating room. When the in vitro splicing of the liver was almost completed, surgeons entirely removed the recipient′s liver and started to transplant the new one. Observation indicators: (1) surgical situations and postoperative recovery of the living donor and the recipient; (2) postoperative pathological examination of the recipient′s liver; (3) follow-up. Follow-up was conducted by outpatient examinations, including monitoring of HCC recurrence, monitoring of new liver function, monitoring and adjustment of immunosuppressive agents, detection of biliary vascular complications, rejection and adverse drug reactions. Regular lifelong follow-up was required for recipients, with the latest follow-up on December 4, 2019. Count data were expressed as absolute numbers or percentages.Results:(1) Surgical situations and postoperative recovery of the living donor and the recipient: operation time, volume of intraoperative blood loss, volume of intraoperative infusion of autologous blood of the living donor were 315 minutes, 200 mL, 200 mL, respectively. The living donor was discharged from hospital on the sixth day after surgery without any complications. The recipient underwent modified piggyback liver transplantation successfully. Graft one was from the right segment free of the middle hepatic vein in the living donor, with a weight of 410 g. Graft two was from the left lateral segment in the donor after brain death, with a weight of 400 g. The graft from donors to recipient weight ratio was 1.2% after splicing. The operation time, duration of anhepatic phase, volume of intraoperative blood loss, volume of intraoperative blood transfusion were 815 minutes, 60 minutes, 1 500 mL, 1 800 mL, respectively. The recipient′s temperature was normal during hospitalization. On the first postoperative day, the level of white blood cell and neutrophilic granulocyte percentage of the recipient reached a peak (17.15×10 9/L and 91.7%, respectively) and then gradually decreased. After anti-infective treatment with piperacillin sodium and sulbactam sodium, both of the two indicators returned to normal on the seventh day after surgery (7.90×10 9/L and 70.9%, respectively), and the antibiotic was discontinued. During the hospitalization, the level of albumin of the recipient fluctuated in 31.0-41.4 g/L, the liver function parameters including total bilirubin, alanine aminotransferase, aspartate aminotransferase, prothrombin time and international normalized ratio gradually returned to normal levels, and the renal function parameters including creatinine and estimated glomerular filtration rate remained within the normal range. On the tenth day after surgery, the recipient was in good condition and discharged from the hospital. (2) Postoperative pathological examination of the recipient′s liver: ① results of the pathological examination showed moderately differentiated HCC with incomplete tumour capsule and no invasion of the liver capsule. The surrounding liver tissues showed hepatitis B-related nodular cirrhosis, and no tumor involvement was detected at the broken end of the hilum. ② The gallbladder presented chronic cholecystitis accompanied by cholesterol deposition, and one abdominal lymph node showed reactive hyperplasia. The immunohistochemical staining showed 10% positive HBsAg and negative HBcAg. (3) Follow-up: the tumor markers of the recipient were tested on November 19, 2019, including α-fetoprotein (2.92 μg/L) and abnormal prothrombin (16 AU/L). Together with the negative result of abdominal colour doppler ultrasound, they collectively indicated no HCC recurrence in the recipient. The liver function parameters including total bilirubin (8.6 μmol/L), alanine aminotransferase (23 IU/L), aspartate aminotransferase (28 IU/L) and albumin (44.0 g/L) of the recipient tested on December 3, 2019, were all in normal levels. Blood concentration of tacrolimus was 4.2 μg/L . The drug dose of mycophenolate mofetil dispersible tablets was adjusted to 250 mg given twice daily, and the drug dose of others was unchanged (tacrolimus 2 mg, once daily; sirolimus 1mg, once daily). No symptoms, signs or examination results indicated biliary vascular complications, rejection or adverse drug reactions. Conclusion:Dual-graft living donor liver transplantation of right segment from an adult living donor combined with a left lateral segment from donation after brain death is safe and effective, which can be used as a suboptimal treatment for patients with HCC beyond Milan criteria.
6.Gut Microbiota Dysbiosis Correlates With Long COVID-19 at One-Year After Discharge
Dongmei ZHANG ; Yaya ZHOU ; Yanling MA ; Ping CHEN ; Jian TANG ; Bohan YANG ; Hui LI ; Mengyuan LIANG ; YuE XUE ; Yao LIU ; Jianchu ZHANG ; Xiaorong WANG
Journal of Korean Medical Science 2023;38(15):e120-
Background:
Long coronavirus disease 2019 (COVID-19) in recovered patients (RPs) is gradually recognized by more people. However, how long it will last and the underlining mechanism remains unclear.
Methods:
We conducted a prospective follow-up study to evaluate the long-term symptoms and clinical indices of RPs at one-year after discharge from Union Hospital, Wuhan, China between December 2020 to May 2021. We also performed the 16S rRNA sequencing of stool samples from RPs and healthy controls (HCs) and analyzed the correlation between the gut microbiota and long COVID-19.
Results:
In total, 187 RPs were enrolled, among them, 84 (44.9%) RPs reported long COVID-19 symptoms at one-year after discharge. The most common long-term symptoms were cardiopulmonary symptoms, including chest tightness after activity (39/187, 20.9%), palpitations on exercise (27/187, 14.4%), sputum (21/187, 11.2%), cough (15/187, 8.0%) and chest pain (13/187, 7.0%), followed by systemic symptoms including fatigue (34/187, 18.2%) and myalgia (20/187, 10.7%), and digestive symptoms including constipation (14/187, 7.5%), anorexia (13/187, 7.0%), and diarrhea (8/187, 4.3%). Sixty-six (35.9%) RPs presented either anxiety or depression (42/187 [22.8%] and 53/187 [28.8%] respectively), and the proportion of anxiety or depression in the long symptomatic group was significantly higher than that in the asymptomatic group (41/187 [50.6%] vs. 25/187 [24.3%]). Compared with the asymptomatic group, scores of all nine 36-Item Short Form General Health Survey domains were lower in the symptomatic group (all P < 0.05). One hundred thirty RPs and 32 HCs (non-severe acute respiratory syndrome coronavirus 2 infected subjects) performed fecal sample sequencing.Compared with HCs, symptomatic RPs had obvious gut microbiota dysbiosis including significantly reduced bacterial diversities and lower relative abundance of short-chain fatty acids (SCFAs)-producing salutary symbionts such as Eubacterium_hallii_group, Subdoligranulum, Ruminococcus, Dorea, Coprococcus, and Eubacterium_ventriosum_group. Meanwhile, the relative abundance of Eubacterium_hallii_group, Subdoligranulum, and Ruminococcus showed decreasing tendencies between HCs, the asymptomatic group, and the symptomatic group.
Conclusion
This study demonstrated the presence of long COVID-19 which correlates with gut microbiota dysbiosis in RPs at one-year after discharge, indicating gut microbiota may play an important role in long COVID-19.
7.Clinical outcome of arthroscopic anatomic single bundle reconstruction of posterior cruciate ligament assisted by internal tension relieving technique combined with rapid rehabilitation in the treatment of posterior cruciate ligament rupture
Yang YU ; Zhenglyu ZHAO ; Bing XIE ; Zhengliang SHI ; Guoliang WANG ; Bohan XIONG ; Ziming GU ; Jinrui LIU ; Yanlin LI
Chinese Journal of Trauma 2023;39(7):593-602
Objective:To compare the clinical efficacies between arthroscopic anatomic single bundle reconstruction of posterior cruciate ligament (PCL) assisted by internal tension relieving technique combined with rapid rehabilitation and anatomic single bundle reconstruction combined with conventional rehabilitation in the treatment of PCL rupture.Methods:A retrospective cohort study was used to analyze the clinical data of 88 patients with PCL rupture admitted to First Affiliated Hospital of Kunming Medical University from September 2016 to September 2020. The patients included 65 males and 23 females, aged 18-55 years [(39.3±10.8)years]. Forty-four patients underwent arthroscopic anatomic single bundle reconstruction of PCL assisted by internal tension relieving technique combined with rapid rehabilitation therapy (tension-relieving group), and 44 patients underwent arthroscopic routine anatomic single bundle reconstruction of PCL combined with conventional rehabilitation (traditional group). The two groups were compared before and at 3, 12, and 24 months after surgery regarding the following items: International Knee Literature Committee (IKDC) score, Hospital for Special Surgery (HSS) score, Lysholm score, knee motion cycle (maximum stride length, minimum stride length, and stride frequency) and 6 kinematic indicators (angle of forward and back extension, angle of internal and external rotation, angle of internal and external rotation, up and down displacement, internal and external displacement, and forward and back displacement). The Marburger arthroscopy score (MAS) and Professor Ao Yingfang′s improved score by secondary arthroscopy were compared between the two groups at 12 months after surgery. The perioperative complications were observed.Results:All patients were followed for 24-36 months [(25.5±6.3)months]. In tension-relieving group and the traditional group, the values of IKDC score were (71.8±9.8)points and (68.5±6.5)points at 3 months after surgery, (87.6±6.0)points and (87.6±5.5)points at 12 months after surgery, and (95.5±3.1)points and (92.8±11.6)points at 24 months after surgery, respectively. The values were gradually increased, significantly higher than those before surgery [(48.1±16.9)points and (47.1±15.0)points] (all P<0.05). There were no significant differences between the two groups at each time point (all P>0.05). In tension-relieving group and the traditional group, the values of HSS score were (74.2±6.2)points and (68.4±9.5)points at 3 months after surgery, (91.9±5.4)points and (88.4±4.7)points at 12 months after surgery, and (97.1±2.0)points and (96.2±2.8)points at 24 months after surgery, respectively. The values of HSS score gradually increased, significantly higher than those before surgery [(57.5±17.7)points and (56.8±14.3)points] (all P<0.05). At 3 and 12 months after surgery, the values of HSS score in the tension-relieving group were significantly higher than those in the traditional group (all P<0.05), but others were not significantly different between the two groups (all P>0.05). In tension-relieving group and the traditional group, the values of Lysholm score were (74.2±14.9)points and (70.3±7.5)points at 3 months after surgery, (90.9±6.1)points and (88.7±4.7)points at 12 months after surgery, and (96.9±3.0)points and (96.3±2.8)points at 24 months after surgery, respectively. The values of Lysholm score were gradually increased, significantly higher than those before operation [(48.7±20.7)points and (48.2±19.9)points] (all P<0.05). There were no significant differences between the two groups at any time points (all P>0.05). At 3, 12, and 24 months after surgery, the motion cycle (maximum stride length, minimum stride length and stride frequency) and 6 kinematic indicators (angle of forward bending and backward extension, angle of internal and external rotation, angle of internal and external rotation, internal and external displacement, up and down displacement, and forward and backward displacement) of knee joint were significantly improved in both groups compared with those before surgery (all P<0.05). At 3, 12, and 24 months after surgery, the forward and backward displacement in the tension-relieving group was significantly decreased than that in the traditional group (all P<0.05), but others were not significantly different between the two groups (all P>0.05). The MAS was rated as excellent to good in 14 patients and fair to poor in 2, with the excellent and good rate of 87.5% (14/16) in the tension-relieving group, while the score was rated as excellent to good in 11 patients and fair to poor in 3, with the excellent and good rate of 78.6% (11/14) in the traditional group ( P>0.05). The Professor Ao Yingfang′s improved score was (10.6±1.5)points in the tension-relieving group, markedly higher than that in the traditional group [(9.6±2.3)points] ( P<0.05). No perioperative complications were observed. Conclusion:Compared with anatomic single bundle reconstruction combined with conventional rehabilitation, arthroscopic anatomic single bundle reconstruction of PCL assisted by internal tension relieving technique combined with rapid rehabilitation have higher subjective function score, better anteroposteric stability during knee movement, and better results of secondary microscopy.
8.Three-month Follow-up Study of Survivors of Coronavirus Disease 2019after Discharge
Limei LIANG ; Bohan YANG ; Nanchuan JIANG ; Wei FU ; Xinliang HE ; Yaya ZHOU ; Wan-Li MA ; Xiaorong WANG
Journal of Korean Medical Science 2020;35(47):e418-
Background:
Most patients including health care workers (HCWs) survived the coronavirus disease 2019 (COVID-19), however, knowledge about the sequelae of COVID-19 after discharge remains limited.
Methods:
A prospectively observational 3-month follow-up study evaluated symptoms, dynamic changes of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) IgG and IgM, lung function, and high resolution computed tomography (HRCT) of survivors of COVID-19 after discharge at Wuhan Union Hospital, China.
Results:
Seventy-six survivors (55 females) with a mean age of 41.3 ± 13.8 years were enrolled, and 65 (86%) were HCWs. A total of 69 (91%) patients had returned to their original work at 3-months after discharge. Most of the survivors had symptoms including fever, sputum production, fatigue, diarrhea, dyspnea, cough, chest tightness on exertion and palpitations in the three months after discharge. The serum troponin-I levels during the acute illness showed high correlation with the symptom of fatigue after hospital discharge (r = 0.782; P = 0.008) and lymphopenia was correlated with the symptoms of chest tightness and palpitations on exertion of patients after hospital discharge (r = −0.285, P = 0.027; r = −0.363, P = 0.004, respectively). The mean values of forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), FEV1/FVC, total lung capacity and diffusion capacity were all normal (> 80% predicted) and lung HRCTs returned to normal in most of the patients (82%), however, 42% of survivors had mild pulmonary function abnormalities at 3-months after discharge. SARS-CoV-2 IgG turned negative in 11% (6 of 57 patients), 8% (4 of 52 patients) and 13% (7 of 55 patients), and SARS-CoV-2 IgM turned negative in 72% (41 of 57 patients), 85% (44 of 52 patients) and 87% (48 of 55 patients) at 1-month, 2-months and 3-months after discharge, respectively.
Conclusion
Infection by SARS-CoV-2 caused some mild impairments of survivors within the first three months of their discharge and the duration of SARS-CoV-2 antibody was limited, which indicates the necessity of long-term follow-up of survivors of COVID-19.
9.Mechanism of NONHSAT248596.1 endogenous competition with miR-146a-5p regulating osteoarthritis cartilage degeneration
Guang YANG ; Yanlin LI ; Guoliang WANG ; Ziwen NING ; Tengyun YANG ; Renjie HE ; Bohan XIONG ; Bing YANG ; Li LI
Chinese Journal of Tissue Engineering Research 2024;28(16):2512-2518
BACKGROUND:Currently,there have been studies on the regulatory mechanism of lncRNA\miRNA\mRNA co-expression network on the occurrence and development of osteoarthritis.Our research group has screened qualified NONHSAT248596.1 and miR-146a-5p through the database in the previous stage,but the corresponding in vivo experiments to verify the above regulatory mechanisms are still lacking. OBJECTIVE:To explore the role of NONHSAT248596.1 in regulating competitive endogenous RNA of miR-146a-5p in cartilage degeneration mediated by stromal cell derived factor type 1/chemokine receptor 4 axis in vivo. METHODS:The models of osteoarthritis were established in 36 New Zealand rabbits by injecting stromal cell derived factor 1 solution into the knee joint of the right hind limb.According to the random number table method,they were divided into four groups.lncRNA group,miRNA group,ceRNA group and control group were injected with lentivirus vector overexpressing NONHSAT248596.1,lentivirus vector overexpressing miR-146a-5p,lentivirus vector overexpressing miR-146a-5p+NONHSAT248596.1 and empty lentivirus vector into the molded knee joint,respectively.At 4,8 and 12 weeks of modeling,cartilage tissues and subchondral bone tissues of the knee joint were taken for relevant detection. RESULTS AND CONCLUSION:Hematoxylin-eosin staining and safranin fast green staining showed different degrees of degeneration in the four groups.At 4 weeks,the cartilage tissue of the lncRNA group showed swelling of chondrocytes,loss of cell polarity,destruction of extracellular matrix,surface erosion,fracture formation and partial or full layer loss of cartilage tissue.The degree of cartilage injury was gradually aggravated with time.The progression of articular cartilage inflammation in the miRNA group was the slowest among the four groups.qRT-PCR showed that at the same time point,mRNA expression levels of NONHSAT248596.1,chemokine receptor 4,matrix metalloproteinase 3,matrix metalloproteinase 9 and matrix metalloproteinase 13 in cartilage tissue of the lncRNA group were higher than those of the other three groups(P<0.05).The mRNA expression levels of miR-146a-5p,aggrecan and type Ⅱ collagen were lower than those of the other three groups(P<0.05).The mRNA expression levels of NONHSAT248596.1,chemokine receptor 4,matrix metalloproteinase 3,matrix metalloproteinase 9 and matrix metalloproteinase 13 in the miRNA group were lower than those in the ceRNA group and control group at 8 and 12 weeks after the model construction(P<0.05).The mRNA expressions of miR-146a-5p,aggrecan and type Ⅱ collagen were higher than those of the ceRNA group and control group(P<0.05).Western blot assay showed that at the same time point,the expression levels of aggrecan and type Ⅱ collagen in cartilage tissue of the lncRNA group were always lower than those of the other three groups(P<0.05).The expression levels of aggrecan and type Ⅱ collagen in cartilage tissue of the miRNA group at 8 and 12 weeks after modeling were higher than those of the ceRNA group and control group(P<0.05).The results showed that miR-146a-5p,as the target of NONHSAT248596.1,could be inhibited by the effect of its ceRNA.After acting on miR-146a-5p,NONHSAT248596.1 regulates the stromal cell derived factor type 1/chemokine receptor 4 axis to affect the expression of matrix metalloprotein,type Ⅱ collagen,and aggrecan in osteoarthritis chondrocytes,resulting in the degradation of extracellular matrix and the loss of proteoglycan.
10.Constructing a model of anterior cruciate ligament reconstruction with autologous Achilles tendon in southern Yunnan small-ear pigs
Bohan XIONG ; Yang YU ; Liling ZHENG ; Tengyun YANG ; Xiaojun LU ; Xu WANG ; Kaiwei LI ; Hong YU ; Yajuan LI ; Kaiyan DONG ; Yaozhang ZHANG ; Jinrui LIU ; Ziming GU ; Bigeng HU ; Yanlin LI
Chinese Journal of Tissue Engineering Research 2024;28(20):3157-3163
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.