Novel strategy of sirolimus plus thymalfasin and huaier granule on tumor recurrence of hepatocellular carcinoma beyond the UCSF criteria following liver transplantation: A single center experience.

Although liver transplantation (LT) lengthens the survival time of patients with hepatocellular carcinoma (HCC), LT patients exhibit a high recurrence rate; particularly those that had advanced HCC associated with the tumor biological characteristics and long-term application of immunosuppressants. A consensus on optimal prophylaxis and treatment for recurrent HCC following LT does not currently exist. The present study retrospectively analyzed data from 36 non-University of California at San Francisco criteria-eligible patients with advanced HCC who underwent LT, and then treated them with sirolimus (SRL)-based therapy with thymalfasin and huaier granules (SRL+, n=18), or with tacrolimus-based therapy (controls; n=18). The SRL+ group had significantly longer recurrence times (P=0.008) and survival times (P<0.0001) (OS, 1-year: 100%, 3-year: 94.4%, 5-year: 77.8%; DFS, 1-year: 88.9%, 3-year: 55.6%, 5-year: 50.0%). Furthermore, compared with pre-LT values and the control group, the SRL+ group had significantly lower serum α-fetoprotein (AFP) levels (both P<0.0001) and percentage of Forkhead box P3 (FoxP3)+ Treg lymphocytes (P<0.001) during the first year. In the SRL+ group, FoxP3+/cluster of differentiation (CD)8+ Treg lymphocyte percentages decreased significantly following LT (P<0.001); however, CD8+/CD3+ T-cells significantly increased (P<0.001). Levels of serum AFP and FoxP3+ Treg cells increased when tumors relapsed, and decreased to near-normal when relapse foci were cured or stabilized. SRL+ therapy may decrease AFP and Treg levels, while increasing CD8+ T cells, indicating an associated mechanism among them. In conclusion, SRL+ therapy appears to be safe and effective in preventing HCC recurrence following LT with no significant adverse events, and warrants further investigation.

Subchondral bone derived mesenchymal stem cells display enhanced osteo-chondrogenic differentiation, self-renewal and proliferation potentials.

Rabbit mesenchymal stem cells (MSCs) are important seed cells in regenerative medicine research, particularly in translational research. In the current study, we showed that rabbit subchondral bone is a reliable source of MSCs. First, we harvested subchondral bone (SCB) from the rabbit knee-joint and initiated the MSC culture by cultivating enzyme-treated SCB. Adherent fibroblast-like cells that outgrew from SCB fulfill the common immuno-phenotypic criteria for defining MSCs, but with low contamination of CD45+ hematopoietic cells. Interestingly, differentiated SCB-MSCs expressed osteogenic and chondrogenic markers at significantly higher levels than those in bone marrow cell suspension-derived MSCs (BMS-MSCs) (P<0.05). No differences in the expression of adipogenic markers between SCB-MSC and BMS-MSC (P>0.05) were observed. Moreover, the results of the colony forming unit-fibroblast assay and sphere formation assay demonstrated that the SCB-MSCs had increased self-renewal potential. SCB-MSCs expressed higher levels of the stemness markers Nanog, OCT4, and Sox-2 compared to in BMS-MSCs (P<0.05). Furthermore, the results of both the CCK-8-based assay and CFSE dilution assay showed that SCB-MSCs exhibited enhanced proliferative capacity. In addition, SCB-MSCs exhibited higher phosphorylation of extracellular signal-related kinase/mitogen-activated protein kinase signaling, which is closely related to MSC proliferation. In conclusion, we identified SCB-MSCs as a novel stem cell population that met the requirements of MSCs; the unique properties of SCB-MSC are important for the potential treatment of tissue damage resulting from disease and trauma.

Radial shockwave treatment promotes human mesenchymal stem cell self-renewal and enhances cartilage healing.

BACKGROUND: Shockwaves and mesenchymal stem cells (MSCs) have been widely accepted as useful tools for many orthopedic applications. However, the modulatory effects of shockwaves on MSCs remain controversial. In this study, we explored the influence of radial shockwaves on human bone marrow MSCs using a floating model in vitro and evaluated the healing effects of these cells on cartilage defects in vivo using a rabbit model. METHODS: MSCs were cultured in vitro, harvested, resuspended, and treated with various doses of radial shockwaves in a floating system. Cell proliferation was evaluated by growth kinetics and Cell Counting Kit-8 (CCK-8) assay. In addition, the cell cycle and apoptotic activity were analyzed by fluorescence activated cell sorting. To explore the "stemness" of MSCs, cell colony-forming tests and multidifferentiation assays were performed. We also examined the MSC subcellular structure using transmission electron microscopy and examined the healing effects of these cells on cartilage defects by pathological analyses. RESULTS: The results of growth kinetics and CCK-8 assays showed that radial shockwave treatment significantly promoted MSC proliferation. Enhanced cell growth was also reflected by an increase in the numbers of cells in the S phase and a decrease in the numbers of cells arrested in the G0/G1 phase in shockwave-treated MSCs. Unexpectedly, shockwaves caused a slight increase in MSC apoptosis rates. Furthermore, radial shockwaves promoted self-replicating activity of MSCs. Transmission electron microscopy revealed that MSCs were metabolically activated by shockwave treatment. In addition, radial shockwaves favored MSC osteogenic differentiation but inhibited adipogenic activity. Most importantly, MSCs pretreated by radial shockwaves exhibited an enhanced healing effect on cartilage defects in vivo. Compared with control groups, shockwave-treated MSCs combined with bio-scaffolds significantly improved histological scores of injured rabbit knees. CONCLUSIONS: In the present study, we found that radial shockwaves significantly promoted the proliferation and self-renewal of MSCs in vitro and safely accelerated the cartilage repair process in vivo, indicating favorable clinical outcomes.

Long-term Outcome of Multiple Small-diameter Drilling Decompression Combined with Hip Arthroscopy versus Drilling Alone for Early Avascular Necrosis of the Femoral Head.

BACKGROUND: Avascular necrosis of femoral head (AVNFH) typically presents in the young adults and progresses quickly without proper treatments. However, the optimum treatments for early stage of AVNFH are still controversial. This study was conducted to evaluate the therapeutic effects of multiple small-diameter drilling decompression combined with hip arthroscopy for early AVNFH compared to drilling alone. METHODS: This is a nonrandomized retrospective case series study. Between April 2006 and November 2010, 60 patients (98 hips) with early stage AVNFH participated in this study. The patients underwent multiple small-diameter drilling decompression combined with hip arthroscopy in 26 cases/43 hips (Group A) or drilling decompression alone in 34 cases/55 hips (Group B). Patients were followed up at 6, 12, and 24 weeks, and every 6 months thereafter. Radiographs were taken at every follow-up, Harris scores were recorded at the last follow-up, the paired t-test was used to compare the postoperative Harris scores. Surgery effective rate of the two groups was compared using the Chi-square test. RESULTS: All patients were followed up for an average of 57.6 months (range: 17-108 months). Pain relief and improvement of hip function were assessed in all patients at 6 months after the surgery. At the last follow-up, Group A had better outcome with mean Harris' scores improved from 68.23 ± 11.37 to 82.07 ± 2.92 (t = -7.21, P = 0.001) than Group B with mean Harris' scores improved from 69.46 ± 9.71 to 75.79 ± 4.13 (t = -9.47, P = 0.037) (significantly different: t = -2.54, P = 0.017). The total surgery effective rate was also significantly different between Groups A and B (86.0% vs. 74.5%; χ2 = 3.69, P = 0.02). CONCLUSION: For early stage of AVNFH, multiple small-diameter drilling decompression combined with hip arthroscopy is more effective than drilling decompression alone.

[Effects of Shock Wave on the Proliferation and Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells].

OBJECTIVE: To explore the effects of the shock wave on the capacity of mesenchymal stem cells(MSCs) to proliferate and differentiate into osteoblasts. METHODS: MSCs were isolated from the bone marrow of healthy donors. The human bone marrow MSCs(BM-MSCs) were divided into 3 groups including blank control group,osteoinduced group and shock wave group. The MSCs in blank control group were cultured with common mediam; the MSCs in osteoinduced group were treated with osteogenic agents and cultured; the MSCs in shock wave group were cultured with common medium and stimulated by shock wave. The morphology of MSCs in each groups were observed by micoscopy; the CCK-8 was used to detect the proliferation ability of MSCs; the alkaline phosphatase staining and von Kossa staining were used to evaluale the differentiation potential of MSCs in each groups. RESULTS: The results of CCK-8 revealed the shock wave could promote cell proliferation as compared with blank control group. The results of alkaline phosphatase and Von Kossa staining showed that the shock wave displayed a stronger ability to promote the human BMMSC differentiation into osteoblasts cells in comparison with the osteoinduced group. The blank control group was weakly positively stainined. CONCLUSION: The shock wave treatment can promote proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.

[Clinical research of popliteal cyst excision combined with debridement of the knee under arthroscopy with local anesthesia].

OBJECTIVE: To evaluate the clinical effects of popliteal cyst excision combined with debridement of the knee under arthroscopy with local anesthesia. METHODS: From February 2009 to August 2014, 52 patients with popliteal cysts were treated in our hospital, including 34 males and 18 females with an average age of 43.6 years old ranging from 14 to 62 years old; 29 cases were on the right knee and 23 on the left knee. Preoperative diagnosis was performed according to MRI findings and to determine whether other knee disorders were associated with the disease, 52 patients were diagnosed as popliteal cyst before operation; 23 cases of simple meniscus injury or cartilage injury, 18 cases of osteoarthritis, 7 cases of cruciate ligament injury, 2 cases of gouty arthritis, 1 case of rheumatoid arthritis, 1 case of pigmented villonodular arthritis. The preoperative clinical manifestations involved knee swelling in 21 cases, knee joint pain in 16 cases, joint lock in 8 cases, leg weakness in 4 cases, and knee joint snapping in 4 cases. Cyst size was 4.0 cm× 3.3 cm to 6.2 cm× 5.3 cm. According to the Rauschning and Lindgren standards, 1 case was grade I, 9 cases were grade II, 42 cases were class III. The rehabilitation plan should be made according to the patient's specific injuries and intraoperative management. During the postoperative follow-up, the postoperative curative effect was evaluated by the standard grading of Rauschning and Lindgren of popliteal cyst. RESULTS: All the incisions healed by first intention, and no complications occurred. All the cases were followed up from 25 to 64 months (averaged 39.6 months). For the Rauschning and Lindgren grading, 43 cases were grade 0, 8 cases were grade I, 1 case were grade II. CONCLUSIONS: Local anesthesia under arthroscopy and popliteal cyst removal knee joint cavity debridement is effective, less trauma, quick recovery, short term effect is good. The recurrence rate is also low because of the primary disease of the knee joint is also treated. After the operation, the rehabilitation plan should be made according to the intraoperative treatment and actively trained so as to recover at an early date.