Enriched Peripheral Blood-Derived Mononuclear Cells for Treating Knee Osteoarthritis.

Osteoarthritis (OA) is a common chronic skeletal disease in the elderly. There is no effective therapy to reverse disease severity and knee OA (KOA) progression, particularly at the late stage. This study aims to examine the effect of peripheral blood-derived mononuclear cells (PBMNCs) on pain and motor function rescue in patients with Kellgren-Lawrence (KL) grade II to IV KOA. Participants received one intra-articular (IA) injection of autologous PBMNCs. The mononuclear cells were isolated from peripheral blood, enriched by a specialized medium (MoFi medium), and separated by Ficoll-Paque solution. The isolated and enriched PBMNCs could differentiate into M1 and M2 macrophages in vitro. The in vivo anti-inflammatory effect of the PBMNCs was similar to that of bone marrow mesenchymal stem cells, evaluated by complete Freund's adjuvant-induced arthritis in rodents. A single-arm and open-label pilot study showed that patients' knee pain and motor dysfunction were significantly attenuated after the cell transplantation, assessed by visual analogue scale (VAS) and Knee injury and Osteoarthritis Outcome Score (KOOS) at 6 and 12 months post-treatment. Notably, the therapeutic effect of the PBMNCs treatment can be stably maintained for 24 months, as revealed by the KOOS scores. These preclinical and pilot clinical data suggest that IA injection of MoFi-PBMNCs might serve as a novel medical technology to control the pain and the progress of KOA.

Induction of type II collagen expression in M2 macrophages derived from peripheral blood mononuclear cells.

The human type II collagen (Col II), specifically expressed in chondrocytes, is a crucial component of the adult hyaline cartilage. We examine the potential of artificial induction of Col II in human peripheral blood mononuclear cells (PBMNCs) as a novel Col II provider. Human PBMNCs were purified and were treated with high doses of macrophage-colony stimulating factor (M-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), or granulocyte-colony stimulating factor (G-CSF) and examined the Col II expression at indicated days. Quantitative Col II expression was validated by real-time reverse transcriptase-polymerase chain reaction (RT-PCR), immunocytochemistry, and flow cytometry. We demonstrate that monocytes in PBMNCs can be artificially induced to express both Col II proteins and M2 macrophage markers by the high concentration of colony-stimulating factors, especially M-CSF and GM-CSF. The Col II proteins were detected on the cell membrane and in the cytoplasm by flow cytometry and immunocytostaining. Combination with IL-4 provided a synergistic effect with M-CSF/GM-CSF to trigger Col II expression in M2 macrophages. These CD206 and Col II double-expressing cells, named modified macrophages, share M2 macrophages' anti-inflammatory potency. We demonstrated that the modified macrophages could significantly attenuate the inflammatory progress of Complete Freund's adjuvant (CFA)-induced arthritis and collagen-induced arthritis in rodents. Here, we provide the first evidence that a modified macrophage population could ectopically express Col II and control the progress of arthritis in animals.

The Effects of Acupressure on Improving Health and Reducing Cost for Patients Undergoing Thoracoscopic Surgery.

OBJECTIVES: This study aimed to assess the effectiveness of practicing acupressure on the Shenmen and Neiguan acupoints with a view to reduce anxiety and improve the comfort and physical health of patients undergoing thoracoscopic surgery. METHODS: A total of 100 hospitalized patients undergoing thoracoscopic surgery were assigned randomly into the experimental (n = 49) and control groups (n = 51). Subjects in the experimental group received routine care plus acupressure on the Shenmen and Neiguan acupoints, while those in the control group received regular routine care. The data were collected using demographic information, physical and surgical data, the Visual Analog Scale (VAS)-A, the State-Trait Anxiety Inventory Y Form (STAI-Y1), and Shortened General Comfort Questionnaire scores. The linear mixed model was used to examine the influences of acupressure on VAS-A and STAI-Y1 scores at different time points before and after the surgery to observe group-by-time interactions. RESULTS: The mean age of the subjects was 60.97 years. All subjects had mild-to-moderate anxiety after surgery and showed a statistically significant decline in regression coefficients on the first and second days after the intervention (ß = -11.61, p = 0.002; ß = -18.71, p < 0.001). Similarly, for STAI-YI scores, the data showed a significant difference in the pre-test and post-test interactions between the two groups (ß = 4.72, p = 0.031). Conversely, acupressure did not have a statistically significant difference on comfort (F = 2.953, p = 0.057). Compared with the control subjects, the experimental subjects used less morphine and developed side effects less frequently (p < 0.01). They were also able to get out of bed after surgery 163.79 min earlier (p < 0.05). CONCLUSIONS: Acupressure is a simple and easy-to-practice treatment. Acupressure on the Shenmen and Neiguan acupoints reduces anxiety and improves recovery in patients after undergoing thoracoscopic surgery.

The effectiveness of acupressure for managing postoperative pain in patients with thoracoscopic surgery: A randomized control trial.

PURPOSE: More than 86% patients experience moderate to severe pain after thoracoscopic surgery. A combination of diverse nonpharmacological pain relief methods is a developing trend for pain management. The purpose of this study was to explore the effect of acupressure in reducing pain after thoracoscopic surgery. DESIGN: A Randomized controlled study with purpose sampling was used for this study. Patients who underwent thoracoscopic surgery at a medical center in central Taiwan were enrolled. Study data was collected from September 2020 to April 2021 after the approval of the institutional review board. A total of 100 participants were randomized into two groups (49 and 51 in the experimental and control groups, respectively). METHODS: Participants in the experimental group received acupressure at the Neiguan (PC6) and Shenmen (HT7) acupoints thrice a day for 2 days, whereas those in the control group received routine treatment and did not receive acupressure. The measurement included questionnaires for the collection of general information, physiological information, and disease rating scale. The Visual Analogue Scale-Pain (VAS-P) was used to measure the severity of pain. SPSS statistical software was used for data analysis. Independent sample t-test and chi-squared test were used for descriptive statistics, and paired t-test and linear mixed model were used to examine the effect of acupressure in alleviating pain. FINDINGS: After acupressure intervention, the pain score of the experimental group was lower than that of the control group, and this difference was significant ß = 17.76, p < 0.001 on day 1 after intervention; ß = 19.80, p < 0.001 on day 2 after intervention. The postoperative pain score in the experimental group on day 2 after intervention was significantly lower than that in the control group (t = 2.039, p = 0.044). After the subjects received acupressure, pain index significantly decreased after considering the interaction between time and group (p < 0.001). Regardless of the type of surgery, there were significant differences in pain index when the interaction between time and group was considered (p < 0.001). CONCLUSIONS: This study provided an experimental basis that acupressure can help in pain management in patients after thoracoscopic surgery, and the pain relief results become more significant as the duration of intervention increases. CLINICAL RELEVANCE: Acupressure is effective in relieving postoperative pain in any type of thoracoscopic surgery. Nurses can use acupressure to help control pain in patients after thoracoscopic surgery.

Differential modulation of methamphetamine-mediated behavioral sensitization by overexpression of Mu opioid receptors in nucleus accumbens and ventral tegmental area.

RATIONALE: Repeated administration of methamphetamine (Meth) induces behavioral sensitization which is characterized by a progressive increase in locomotor response after each injection. Previous studies have shown that Mu opioid receptors (MORs) can regulate Meth-mediated behavioral sensitization. However, the reported interactions are controversial; systemic activation of MORs either enhanced or suppressed Meth sensitization. It is possible that alteration of Meth sensitization after systemic administration of MOR ligands reflects the sum of distinct MOR reactions in multiple brain regions. OBJECTIVES: The purpose of the present study was to examine the actions of MORs on Meth sensitization after regionally selective overexpression of human MOR through an AAV6-based gene delivery system. METHOD: We demonstrated that adeno-associated virus (AAV)-MOR increased MOR immunoreactivity and binding in vitro. AAV-MOR or AAV-green fluorescent protein (GFP) was injected into the nucleus accumbens (NAc) or ventral tegmental area (VTA) of adult mice. Two weeks after viral infection, animals received Meth or saline for five consecutive days. Locomotor behavior and striatal dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) level were determined. RESULTS: Repeated administration of Meth progressively increased locomotor activity; this sensitization reaction was attenuated by intra-NAc AAV-MOR microinjections. Infusion of AAV-MOR to VTA enhanced Meth sensitization. AAV-MOR significantly enhanced DA levels in VTA after VTA infection but reduced DOPAC/DA turnover in the NAc after NAc injection. CONCLUSION: Our data suggest a differential modulation of Meth sensitization by overexpression of MOR in NAc and VTA. Regional manipulation of MOR expression through AAV may be a novel approach to control Meth abuse and psychomimetic activity.

Delayed treatment with a p53 inhibitor enhances recovery in stroke brain.

OBJECTIVE: Cerebral ischemia can activate endogenous reparative processes, such as proliferation of endogenous neural progenitor cells (NPCs) in the subventricular zone (SVZ). Most of these new cells die shortly after injury. The purpose of this study was to examine a novel strategy for treatment of stroke at 1 week after injury by enhancing the survival of ischemia-induced endogenous NPCs in SVZ. METHODS: Adult rats were subjected to a 90-minutes middle cerebral artery occlusion. A p53 inhibitor pifithrin-alpha (PFT-alpha) was administered to stroke rats from days 6 to 9 after middle cerebral artery occlusion. Locomotor behavior was measured using an activity chamber. Proliferation, survival, migration, and differentiation of endogenous NPCs were examined using quantitative reverse transcription polymerase chain reaction, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling, and immunohistochemistry. RESULTS: PFT-alpha enhanced functional recovery as assessed by a significant increase in multiple behavioral measurements. Delayed PFT-alpha treatment had no effect on the cell death processes in the lesioned cortical region. However, it enhanced the survival of SVZ progenitor cells, and promoted their proliferation and migration. PFT-alpha inhibited the expression of a p53-dependent proapoptotic gene, termed PUMA (p53-upregulated modulator of apoptosis), within the SVZ of stroke animals. The enhancement of survival/proliferation of NPCs was further found in SVZ neurospheres in tissue culture. PFT-alpha dose-dependently increased the number and size of new neurosphere formation. INTERPRETATION: Delayed treatment with a p53 inhibitor PFT-alpha is able to modify stroke-induced endogenous neurogenesis and improve the functional recovery in stroke animals.

Mesencephalic astrocyte-derived neurotrophic factor reduces ischemic brain injury and promotes behavioral recovery in rats.

Mesencephalic astrocyte-derived neurotrophic factor (MANF), also known as arginine-rich, mutated in early stage of tumors (ARMET), is a secreted protein that reduces endoplasmic reticulum (ER) stress. Previous studies have shown that MANF mRNA expression and protein levels are increased in the cerebral cortex after brain ischemia, a condition that induces ER stress. The function of MANF during brain ischemia is still not known. The purpose of this study was to examine the protective effect of MANF after ischemic brain injury. Recombinant human MANF was administrated locally to the cerebral cortex before a 60-min middle cerebral artery occlusion (MCAo) in adult rats. Triphenyltetrazolium chloride (TTC) staining indicated that pretreatment with MANF significantly reduced the volume of infarction at 2 days after MCAo. MANF also attenuated TUNEL labeling, a marker of cell necrosis/apoptosis, in the ischemic cortex. Animals receiving MANF pretreatment demonstrated a decrease in body asymmetry and neurological score as well as an increase in locomotor activity after MCAo. Taken together, these data suggest that MANF has neuroprotective effects against cerebral ischemia, possibly through the inhibition of cell necrosis/apoptosis in cerebral cortex.

9-Cis-retinoic acid reduces ischemic brain injury in rodents via bone morphogenetic protein.

Retinoic acid (RA), a biologically active derivative of vitamin A, has protective effects against damage caused by H(2)O(2) or oxygen-glucose deprivation in mesangial and PC12 cells. In cultured human osteosarcoma cells, RA enhances the expression of bone morphogenetic protein-7 (BMP7), a trophic factor that reduces ischemia- or neurotoxin-mediated neurodegeneration in vivo. The purpose of this study is to examine whether RA reduces ischemic brain injury through a BMP7 mechanism. We found that intracerebroventricular administration of 9-cis-retinoic acid (9cRA) enhanced BMP7 mRNA expression, detected by RT-PCR, in rat cerebral cortex at 24 hr after injection. Rats were also subjected to transient focal ischemia induced by ligation of the middle cerebral artery (MCA) at 1 day after 9cRA injection. Pretreatment with 9cRA increased locomotor activity and attenuated neurological deficits 2 days after MCA ligation. 9cRA also reduced cerebral infarction and TUNEL labeling. These protective responses were antagonized by the BMP antagonist noggin given 1 day after 9cRA injection. Taken together, our data suggest that 9cRA has protective effects against ischemia-induced injury, and these effects involve BMPs.

The three-item clock-drawing test: a simplified screening test for Alzheimer's disease.

The Clock-Drawing Test (CDT) has been used to screen for Alzheimer's disease (AD) as a supplement to cognitive tests that focus on memory impairment. We examined a comprehensive scoring system of the CDT in screening of AD in a Chinese population and derived a simplified scoring system. All 403 (144 AD and 259 nondemented) subjects were administered the CDT, including both the drawing part (CDT-D) and the copying part (CDT-C). The Cognitive Abilities Screening Instrument and the Clinical Dementia Rating were also administered. Stepwise discriminant analysis was used to develop a simplified CDT scoring system. The optimal CDT cutoff scores (CDT-D: 10/11; CDT-C: 12/13) show intermediate sensitivity (CDT-D: 66.7%; CDT-C: 51.4%) and specificity (CDT-D: 74.5%; CDT-C: 74.1%). The simplified 3-item CDT scoring system, with a cutoff score of 2/3, has a sensitivity of 72.9% and a specificity of 65.6%; it can be used as a quick test for AD screening.