BACKGROUNDSeveral reports have shown the progression of articular cartilage degeneration after anterior cruciate ligament (ACL) reconstruction. No report has been published about the cartilage comparing changes after single-bundle (SB) and double-bundle (DB) ACL reconstructions. The purpose of this study was to evaluate the articular cartilage changes after SB and DB ACL reconstructions by second-look arthroscopy.

METHODSNinety-nine patients who received arthroscopic ACL reconstruction were retrospectively reviewed at an average of 14 months after reconstruction, 58 patients underwent SB ACL reconstruction and 41 patients underwent DB ACL reconstruction. Hamstring tendon autografts were used in all patients. Second-look arthroscopy was done in conjunction with the tibial staple fixation removal at least one year after the initial ACL reconstruction. Arthroscopic evaluation and grading of the articular cartilage degeneration for all patients were performed at the initial ACL reconstruction, and at the second-look arthroscopy.

RESULTSThe average cartilage degeneration at the patellofemoral joint (PFJ) was found significantly worsened after both SB and DB ACL reconstructions. This worsening were not seen at medial tibiofemoral joint (TFJ) and lateral TFJ. Grade II cartilage damage was the most common. At second-look arthroscopy, the average patellar cartilage degeneration was 1.14 ± 0.14 (at first look 0.52 ± 0.11) for the SB group, and 1.22 ± 0.15 (at first look 0.56 ± 0.12) for the DB group. The average trochlear cartilage degeneration was 1.05 ± 0.16 (at fist look 0.10 ± 0.06) and 0.66 ± 0.17 (at fist look 0.17 ± 0.09), respectively. The average patellar cartilage degeneration showed no significant difference in both groups. However, the average trochlea cartilage degeneration in DB group was significantly less than in SB group.

CONCLUSIONSPatellofemoral cartilage degeneration continued to aggravate after ACL reconstruction. DB ACL reconstruction could significantly decrease the trochlea cartilage degeneration compared with SB ACL reconstruction.

Adolescent ; Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Reconstruction ; methods ; Arthroscopy ; methods ; Cartilage, Articular ; surgery ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Second-Look Surgery ; methods ; Treatment Outcome ; Young Adult

BACKGROUNDThe proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood (PB-MSCs) were investigated under hypoxia and serum deprivation conditions in vitro so as to evaluate the feasibility for autologous PB-MSCs applications in cartilage repair.

METHODSMSCs were mobilized into peripheral blood by granulocyte colony stimulating factor (G-CSF) and AMD3100. The blood samples were collected from central ear artery of rabbits. Adhered cells were obtained by erythrocyte lysis buffer and identified as MSCs by adherence to plastic, spindle shaped morphology, specific surface markers, differentiation abilities into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. MSCs were cultured in four groups at different oxygen tension (20% O2 and 2% O2), with or without 10% fetal bovine serum (FBS) conditions: 20% O2 and 10% FBS complete medium (normal medium, N), 20% O2 and serum deprivation medium (D), 2% O2 and 10% FBS complete medium (hypoxia, H), 2% O2 and serum deprivation (HD). Cell proliferation was determined by CCK-8 assay. Apoptosis was detected by Annexin V/PI and terminal deoxynucleotide transferase dUTP nick end labeling (TUNEL) staining.

RESULTSSpindle-shaped adherent cells were effectively mobilized from peripheral blood by a combined administration of G-CSF plus AMD3100. These cells showed typical fibroblast-like phenotype similar to MSCs from bone marrow (BM-MSCs), and expressed a high level of typical MSCs markers CD29 and CD44, but lacked in the expression of hematopoietic markers CD45 and major histocompatibility complex Class II (MHC II). They could also differentiate into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. No significant morphological differences were found among the four groups. It was found that hypoxia could enhance proliferation of PB-MSCs regardless of serum concentration, but serum deprivation inhibited proliferation at the later stage of culture. Apart from that, hypoxia or serum deprivation could promote the apoptosis of PB-MSCs after 48 hours; the effect was stronger when these two conditions combined together. Furthermore, the effect of serum deprivation on apoptosis was stronger compared with that of hypoxia.

CONCLUSIONSPB-MSCs possess similar phenotypes as BM-MSCs. Their differentiation and proliferation abilities make them a new source of seed cells for ischemia-related cell therapy and tissue engineering in the field of the articular cartilage repair.

Animals ; Apoptosis ; physiology ; Cell Hypoxia ; physiology ; Cell Proliferation ; Cells, Cultured ; In Situ Nick-End Labeling ; Mesenchymal Stromal Cells ; cytology ; Rabbits

Objective To establish a liquid biopsy technique of KRAS gene G12D mutation and to assess its diagnostic value. Methods KRAS G12D mutation was analyzed by ddPCR in plasma DNA from 52 colorectal cancer patients and compared that of to 80 healthy subjects. KRAS gene sequencing in cancerous tissue of colorectal cancer patient being set as a golden standard, we evaluated the accuracy of ddPCR and analyzed the correlation between G12D mutation rate, plasma concentration;and their clinical manifestations in CRC. Results ddPCR indicated that KRAS G12D mutation rate and concentration(26.92％, 81.5 copies/mL) in the plasma samples of colorectal cancer patients were significantly higher than that of healthy subjects (8.75％, 16 copies/mL). Colorectal cancer patients with highly differentiated adenocarcinoma showed a significantly higher number of mutant copies than medium and low differentiated adenocarcinoma(P＜0.05);M2 patients had a significantly higher number of mutant copies than N1 and NO patients (P＜0.05);The concordance rate of KRAS gene mutation between cancerous tissue and plasma ctDNA was 87.50％ in CRC.Conclusions ddPCR is a fast, noninvasive and accurate method for plasma testing of ctDNA, and the test results could be used to monitor the course of the disease and as clinical guidelines.

Antirheumatic Agents ; adverse effects ; Dermatitis, Exfoliative ; Humans ; Hydroxychloroquine ; adverse effects ; Lupus Erythematosus, Systemic ; drug therapy ; Psoriasis ; chemically induced

Adipose tissue is a promising target for treating obesity and metabolic diseases. However, pharmacological agents usually fail to effectively engage adipocytes due to their extraordinarily large size and insufficient vascularization, especially in obese subjects. We have previously shown that during cold exposure, connexin43 (Cx43) gap junctions are induced and activated to connect neighboring adipocytes to share limited sympathetic neuronal input amongst multiple cells. We reason the same mechanism may be leveraged to improve the efficacy of various pharmacological agents that target adipose tissue. Using an adipose tissue-specific Cx43 overexpression mouse model, we demonstrate effectiveness in connecting adipocytes to augment metabolic efficacy of the β ₃-adrenergic receptor agonist Mirabegron and FGF21. Additionally, combing those molecules with the Cx43 gap junction channel activator danegaptide shows a similar enhanced efficacy. In light of these findings, we propose a model in which connecting adipocytes via Cx43 gap junction channels primes adipose tissue to pharmacological agents designed to engage it. Thus, Cx43 gap junction activators hold great potential for combination with additional agents targeting adipose tissue.