OBJECTIVETo discuss a new technique about reconstruction of medial collateral ligament (MCL) with double bundle allograft and to evaluate the short-term clinical efficacy.

METHODSAll 53 patients who suffered from valgus instability of the knee were selected. All cases were diagnosed of MCL injury because the medial gap of the knee widened more than 5 mm compared with collateral knee by the stress X-ray, MRI displayed discontinuity of MCL and valgus stress test was positive. All patients were accepted arthroscopic evaluation through inferomedial and inferolateral arthroscopy portal incisions to ascertain whether there were intra-articular injuries. An 8 cm incision was made from 1 cm superior adductor tubercle to 5 cm proximal medial tibia joint line in a longitudinal fashion. The anterior tibia insertion was defined as 15 mm lateral from the medial tibia edge and 45 mm under the medial tibia joint line. The posterior tibia insertion was defined as 15 mm lateral from the medial tibia edge and 20 mm under the medial tibia joint line. We used 5 mm or 6 mm reamer to drill the tibia tunnel along with guide pin, and then drill the femur tunnel with 6 mm or 7 mm drill in the top of the adductor tubercle about 25 mm or 30 mm length. The allograft was pulled into the tunnel from tibia to the femur and fixed with absorbable interference screw. Patients carried out active rehabilitation program after operation. One year after the operation, IKDC score, Lysholm score were used to evaluate the clinical effect.

RESULTSThe IKDC score (A or B, 86.78% vs. 0), Lysholm scores (89.7 ± 3.4 vs. 51.8 ± 4.9, t = -79.724, P < 0.05) were significantly improved compared with preoperative in all patients. Medial joint widened gap decreased from (10.4 ± 2.4) mm preoperative to (2.8 ± 1.5) mm postoperative from X ray and the differences were significant (t = 41.727, P < 0.05). Among these patients, the medial joint widened gap of 46 cases were less than 3 mm, 7 cases were from 3 mm to 5 mm. The range of motion was 135.4° ± 2.5° preoperative and 132.7° ± 3.7° postoperative. The 9 patients still had medial tenderness 1 year after operation.

CONCLUSIONApplication double bundle allograft technique to reconstruct MCL can significantly improve the stability of the knee and the short-term clinical efficacy was sure.

Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Medial Collateral Ligament, Knee ; injuries ; surgery ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Transplantation, Homologous ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the relationship of chimerism status of cell subsets with engraftment, occurrence of acute graft versus host disease (aGVHD), graft rejection and disease relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSChimerism status in peripheral blood (PB) and bone marrow (BM) of 65 patients received allo-HSCT were monitored at regular intervals post-transplant. Fluorescence-activated cell sorter (FACS) was used to sort CD3(+)T lymphocytes in 65 cases, CD3(-)CD56(+)CD16(+)NK cells in 52 cases, CD15(+) granulocytes in 32 cases and CD19(+)B lymphocytes in 20 cases post transplants. The chimerism status of different lineage cells was analyzed by polymerase chain reaction amplification of short tandem repeats (PCR-STR).

RESULTSOn day +7, NK-cells donor chimerism (DC 55.5%) was higher than other cell subsets. T lymphocyte was the latest one to reach complete donor chimerism (CDC) with a median on day +21. Patients whose T lymphocytes donor chimerism was more than 70% on day +7 and more than 95% on day +14 had a high risk for acute aGVHD. In all cases except those with ALL, the decreased DC of T lymphocytes were observed before molecular or hematological relapse occurred.

CONCLUSIONSerial and quantitative T cell chimerism analysis provides a reliable and rapid screening method for the early detection of engraftment, graft rejection, disease relapse and occurrence of aGVHD, therefore, is a prognostic tool to identify patients at high risk of aGVHD and disease relapse following allo-HSCT.

Adolescent ; Adult ; Child ; Chimerism ; Female ; Graft Rejection ; immunology ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Recurrence ; T-Lymphocytes ; immunology ; Young Adult

Aim To explore and verify the possible mechanism of Jiawei Duhuo Jisheng Mixture(JDJM)in the treatment on Knee Osteoarthritis(KOA)via using network pharmacology and animal experiment. Methods The ingredients of JDJM and relevant targets were collected from TCMSP and BATMAN-TCM database. The KOA-related targets were collected from GeneCard, OMIM and GEO databases. The common targets were acquired by intersecting ingredients-related and KOA-related targets, and then the Ingredient-Disease-Target Network and PPI network were constructed by Cytoscape 3.7.2 software and STRING platform. GO and KEGG enrichment analysis were performed based on Metascape database. Finally, the key targets and relevant mechanism were validated via animal experiment. Results In the network pharmacology study, 180 active ingredients related to treatment on KOA by JDJM were collected, and 152 common targets were confirmed. PPI network analysis showed that AKT1 might be the key targets of JDJM in the treatment on KOA. GO and KEGG enrichment analysis revealed that the key target mainly concentrated on inflammatory response and apoptosis. Animal experiment confirmed that JDJM could improve lesion in KOA rabbits, and suppress the expression levels of IL-1β, TNF-α, Caspase 3 and BAX in serum and articular fluid. AKT1 expression(including mRNA and protein)in articular cartilage was also down-regulated. Conclusions Based on the results of network pharmacology and animal experiment, JDJM may relieve KOA severity by anti-inflammatory and anti-apoptotic effects through a variety of molecular signaling pathways.

This study aimed to investigate the effect of methyl eugenol(ME) on hypoxia/reoxygenation(H/R)-induced injury of human renal tubular epithelial HK-2 cells and its mechanism. The viability of HK-2 cells cultured with different concentrations of ME and exposed to H/R was detected by cell counting kit-8(CCK-8) assay. The effect of ME on the morphology of HK-2 cells was observed under an inverted microscope. The content of intracellular reactive oxygen species in different groups was detected after 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA) fluorescence staining. Cell apoptosis was determined by flow cytometry. Changes in mitochondrial membrane potential were monitored by JC-1 dye. The concentrations of nuclear factor erythroid 2 related factor 2(Nrf2), heme oxygenase-1(HO-1), and nicotinamide adenine dinucleotide phosphatase oxidase 4(Nox4) were measured by Western blot, followed by the assay of Nrf2 concentration changes in cytoplasm and nucleus by confocal fluorescence staining. The results showed that when the concentration of ME was 0-40 μmol·L~(-1), the activity of HK-2 cells was not affected. Compared with the model group, ME enhanced the activity of HK-2 cells and the cell morphology was normal. As revealed by further experiments, ME inhibited the release of reactive oxygen species and the decline in mitochondrial membrane potential of HK-2 cells after H/R injury, promoted Nrf2/HO-1 expression and Nrf2 translocation to the nucleus, and down-regulated the expression of Nox4, thereby significantly reducing apoptosis. This protective effect of ME could be reversed by the specific Nrf2 inhibitor ML385. These findings have preliminarily proved that ME effectively protected HK-2 cells against H/R injury, which might be related to its promotion of Nrf2/HO-1 signaling pathway and inhibition of Nox4. Such exploration on the possible mechanism of ME in the treatment of renal ischemia-reperfusion injury(IRI) and protection of organ function from the perspective of antioxidant stress has provided reference for related research on the treatment of acute kidney injury with traditional Chinese medicine.
Apoptosis ; Epithelial Cells/metabolism* ; Eugenol/pharmacology* ; Heme Oxygenase-1/metabolism* ; Humans ; Hypoxia ; NF-E2-Related Factor 2/metabolism* ; Oxidative Stress ; Reactive Oxygen Species ; Reperfusion Injury/drug therapy*