Clinical significance, molecular characterization, and immune microenvironment analysis of coagulation-related genes in clear cell renal cell carcinoma.

Background: Numerous studies have revealed a tight connection between tumor development and the coagulation system. However, the effects of coagulation on the prognosis and tumor microenvironment (TME) of clear cell renal cell carcinoma (ccRCC) remain poorly understood. Methods: We employed the consensus clustering method to characterize distinct molecular subtypes associated with coagulation patterns. Subsequently, we examined variations in the overall survival (OS), genomic profiles, and TME characteristics between these subtypes. To develop a prognostic coagulation-related risk score (CRRS) model, we utilized the least absolute shrinkage and selection operator Cox regression and stepwise multivariate Cox regression analyses. We also created a nomogram to aid in the clinical application of the risk score, evaluating the relationships between the CRRS and the immune microenvironment, responsiveness to immunotherapy, and targeted treatment. The clinical significance of PLAUR and its biological function in ccRCC were also further analyzed. Results: There were significant differences in clinical features, prognostic stratification, genomic variation, and TME characteristics between the two coagulation-related subtypes. We established and validated a CRRS using six coagulation-related genes that can be employed as an effective indicator of risk stratification and prognosis estimation for ccRCC patients. Significant variations in survival outcomes were observed between the high- and low-risk groups. The nomogram was proficient in predicting the 1-, 3-, and 5-year OS. Additionally, the CRRS emerged as a novel tool for evaluating the clinical effectiveness of immunotherapy and targeted treatments in ccRCC. Moreover, we confirmed upregulated PLAUR expression in ccRCC samples that was significantly correlated with poor patient prognosis. PLAUR knockdown notably inhibited ccRCC cell proliferation and migration. Conclusion: Our data suggested that CRRS may be employed as a reliable predictive biomarker that can provide therapeutic benefits for immunotherapy and targeted therapy in ccRCC.

Manual Therapy in Primary Dysmenorrhea: A Systematic Review and Meta-Analysis.

Objective: This research aimed to assess the effectiveness of manual therapy in alleviating pain among women undergoing primary dysmenorrhea (PD). Methods: All randomized controlled trials (RCTs) regarding manual therapy for PD were searched from online databases, spanning from their inception to July 2023. The identified literature underwent a thorough screening process, and the data were meticulously extracted and analyzed using RevMan 5.3. Subsequently, the included studies underwent Cochrane's quality assessment and meta-analysis. The evidence obtained was then assessed using the grading of recommendations, assessment, development, and evaluation (GRADE) approach. Results: 32 RCTs, involving 2566 women were finally included for analysis. The overall quality of the concluding evidence was generally rated as low or very low. Performance bias and blind bias were found to be the main risk of bias of the included studies. In comparison to no treatment, manual therapy demonstrated a significant increase in pain relief in short-term (n=191, MD=1.30, 95% CI: 0.24~2.37). The differences in the effects of manual therapy and the placebo on pain intensity may not be statistically significant (n=255, MD=0.10, 95% CI: -0.37~0.58). In contrast to NSAIDs, manual therapy exhibited superior pain alleviation (n=507, MD=3.01, 95% CI: 1.08~4.94) and a higher effective rate (n=1029, OR=4.87, 95% CI: 3.29~7.20). Importantly, no severe adverse events were reported across all studies, indicating a relatively safe profile for manual therapy. Conclusion: Manual therapy presented promise in effectively relieving menstrual pain with minimal adverse events in short term, outperforming both no treatment and NSAIDs. However, this conclusion is tempered by the low quality of the included RCTs, highlighting the necessity for more robust trials to validate it.

Exploring the association between Frailty Index and low back pain in middle-aged and older Chinese adults: a cross-sectional analysis of data from the China Health and Retirement Longitudinal Study (CHARLS).

OBJECTIVES: This study explored the association between the Frailty Index (FI) and low back pain (LBP) in middle-aged and older Chinese adults. We hypothesised that a higher FI correlates with increased LBP prevalence. DESIGN: Cross-sectional analysis. SETTING: The study used data from the China Health and Retirement Longitudinal Study (CHARLS) across various regions of China. PARTICIPANTS: The analysis included 6375 participants aged 45 and above with complete LBP and FI data from the CHARLS for 2011, 2013 and 2015. We excluded individuals under 45, those with incomplete LBP data, participants with fewer than 30 health deficit items and those missing covariate data. OUTCOME MEASURES: We constructed an FI consisting of 35 health deficits. Logistic multivariable regression examined the relationship between FI and LBP, using threshold analysis to identify inflection points. Sensitivity analyses were performed to ensure the robustness of the findings. RESULTS: Of the participants, 27.2% reported LBP. A U-shaped association was observed between FI and LBP, with the highest quartile (Q4, FI ≥0.23) showing more than a twofold increased risk of LBP (OR=2.90, 95% CI: 2.45-3.42, p<0.001). Stratified analysis showed a significant association in participants under 60, particularly in the lowest FI quartile (OR=1.43, 95% CI: 1.14 to 1.79). Sensitivity analysis upheld the robustness of the primary results. CONCLUSIONS: The findings suggest a complex relationship between frailty and LBP, highlighting the need for early screening and tailored interventions to manage LBP in this demographic. Further research is necessary to understand the mechanisms of this association and to validate the findings through longitudinal studies.

Effect of Traditional Chinese Non-Pharmacological Therapies on Knee Osteoarthritis: A Narrative Review of Clinical Application and Mechanism.

Knee osteoarthritis (KOA) stands as a degenerative ailment with a substantial and escalating prevalence. The practice of traditional Chinese non-pharmacological therapy has become a prevalent complementary and adjunctive approach. A mounting body of evidence suggests its efficacy in addressing KOA. Recent investigations have delved into its underlying mechanism, yielding some headway. Consequently, this comprehensive analysis seeks to encapsulate the clinical application and molecular mechanism of traditional Chinese non-pharmacological therapy in KOA treatment. The review reveals that various therapies, such as acupuncture, electroacupuncture, warm needle acupuncture, tuina, and acupotomy, primarily target localized knee components like cartilage, subchondral bone, and synovium. Moreover, their impact extends to the central nervous system and intestinal flora. More perfect experimental design and more comprehensive research remain a promising avenue in the future.

Short-term and long-term effectiveness of acupuncture and Tuina on knee osteoarthritis: study protocol for a randomized controlled trial.

Background: The effectiveness of acupuncture and tuina in treating knee osteoarthritis (KOA) is still controversial, which limits their clinical application in practice. This study aims to evaluate the short-term and long-term effectiveness of acupuncture and tuina on KOA. Methods/design: This parallel-group, multicenter randomized clinical trial (RCT) will be conducted at the outpatient clinic of five traditional Chinese medicine hospitals in China. Three hundred and thirty participants with KOA will be randomly assigned to acupuncture, tuina, or home-based exercise group with a ratio of 1:1:1. The primary outcome is the proportion of participants achieving a minimal clinically important improvement defined as a ≥ 12% reduction on the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain dimension on short term (week 8) and long term (week 26) compared with baseline. Secondary outcomes are knee joint conditions (pain, function, and stiffness), self-efficacy of arthritis, quality of life, and psychological conditions, which will be evaluated by the WOMAC score and the Patient Global Assessment (PGA), and in addition, the respondents index of OMERACT-OARSI, Short Form 12 Health Survey (SF-12), arthritis self-efficacy scale, and European five-dimensional health scale (EQ-5D). Adverse events will be collected by self-reported questionnaires predefined. Clinical trial registration: https://www.chictr.org.cn.

Development of CORE-CM core outcome domain sets for trials of Chinese medicine for lumbar spinal stenosis.

OBJECTIVES: Most Asian countries have employed Chinese medicine (CM) and Western medicine to treat lumbar spinal stenosis (LSS). Evidence synthesis and comparison of effectiveness are difficult since outcomes examined and presented through trials possess heterogeneity. This study aimed to solve the outcome problems for CM clinical trials in LSS by building a core outcome set (COS). METHODS: To achieve an agreement on a set of core outcome domains, a four-phase study was carried out. First, we identified candidate outcome domains by systematically reviewing trials. In addition, we identified outcome domains associated with patients by conducting semistructured interviews with patients. Next, outcome domains were processed through a national two-round Delphi survey, in which 18 patients and 21 experts were recruited. Finally, the above domains were converted as a core outcome domain set based on a consensus meeting, in which 24 stakeholders were recruited. RESULTS: Seventeen outcome subdomains were identified by the systematic review and interviews. The Delphi survey assigned a priority to four outcome domains in the first round and four outcomes additionally in the second round. The core outcome domains were determined through discussion and redefinition of outcomes in the consensus meeting: pain and discomfort, health-related quality of life, lumbar function, activities of daily living, measures of walking, patient global assessment, adverse events and CM-specific outcomes. CONCLUSION: COS-CM-LSS is likely to enhance the consistency of outcomes reported in clinical trials. In-depth research should be conducted for the exploration of the best methods to examine the above outcomes.

Non-pharmaceutical Chinese medical therapies for degenerative lumbar spinal stenosis: A systematic review and meta-analysis of randomized controlled trials.

OBJECTIVE: The objective of the study was to assess the effectiveness of utilizing Non-Pharmaceutical Chinese Medical (NPCM) therapy singularly or in combination for the treatment of Degenerative Lumbar Spinal Stenosis (DLSS). METHODS: The comprehensive search for all randomized controlled trials regarding NPCM therapies for the treatment of DLSS was performed through online databases searches, commencing from their inception to January 1st, 2023. The relevant literature underwent a thorough screening process, and the data was meticulously extracted and subjected to analysis through the implementation of RevMan 5.3 software. The Cochrane Risk of Bias tool was employed to assess the potential risk of bias. The synthesis of evidence was performed Grading of Recommendations Assessment, Development, and Evaluation. RESULTS: The extensive search procedure produced 5674 records, including data from 37 studies of 38 comparisons (2965 participants). Moderate evidence was obtained demonstrating that the application of acupuncture for a duration of 6-8 weeks was significantly superior to sham acupuncture in terms of intermediate-term (6 months) alleviation of back pain (2 trials, n = 128; MD, -1.08; 95% CI, -1.81∼-0.34) and improvement in lumbar function (2 trials, n = 128; MD, -1.40; 95% CI, -2.93∼-0.13). The available low evidence suggested that, as compared to sham acupuncture, acupuncture was effective in reducing short-term (3 months) back pain and enhancing lumbar function but had no impact on leg pain. A trial with low risk of bias found that acupuncture was more effective than sham acupuncture in enhancing disability and walking capabilities. The other studies presented inconsistent evidence with regards to the efficacy of the various interventions employed. CONCLUSIONS: Evidence of low-to-moderate quality suggests that for DLSS patients, the implementation of acupuncture in comparison to sham acupuncture presents favorable outcomes in terms of short- and intermediate-term alleviation of back pain, improvement in lumbar function, enhancement of disability and walking capacity. The conclusion regarding the efficacy of other NPCM therapies was not obtained due to the insufficient quality of the available evidence. REGISTRATION: PROSPERO CRD42022307631.

A Protocol for a Single-Centered, Pragmatic, Randomized, Controlled, Parallel Trial Comparing Comprehensive Nonsurgical Therapy Options for Individuals with Lumbar Spinal Stenosis.

Aim: Lumbar spinal stenosis (LSS) is a long-term degenerative disease. Considering the risks and advantages of the patient's age range and the characteristics of the condition, non-surgical treatment is recommended. To determine the best first-line non-surgical therapy for LSS, few studies have examined different non-surgical therapies. Therefore, the main objective of this study is to determine whether the selection of comprehensive Chinese medicine (CM) treatment for LSS is more successful than non-surgical conservative treatment. Patients and Methods: In this two-armed, parallel, single-centered, pragmatic randomized controlled study, 94 LSS participants will be randomized to receive 24 sessions of comprehensive CM therapy or conservative treatment for 3 months, with follow-up assessments at 6, 9, 12, and 15 months. The primary outcome will be based on the success rate of the Zurich Claudication Questionnaire (ZCQ) for the most clinical important difference (MCID) at 3 and 15 months. Secondary outcomes include Numerical Rating Scale (NRS) scores for back and leg pain, ZCQ scores, Oswestry Disability Index scores for lumbar dysfunction, and Short-Form 12 scores for health-related quality of life at 3, 6, 9, 12, and 15 months. Adverse events and incidences of surgery will be reported anytime during the trial and follow-up. Conclusion: This protocol examines the comparative efficacy of comprehensive CM therapy compared with conventional care through a pragmatic randomized controlled trial to present data to facilitate clinical or policy decision-making. The outcomes will make it easier to decide which patient-centered treatments to prioritize for LSS.

Tuina for peripherally-induced neuropathic pain: A review of analgesic mechanism.

Peripherally-induced neuropathic pain (pNP) is a kind of NP that is common, frequent, and difficult to treat. Tuina, also known as massage and manual therapy, has been used to treat pain in China for thousands of years. It has been clinically proven to be effective in the treatment of pNP caused by cervical spondylosis, lumbar disc herniation, etc. However, its analgesic mechanism is still not clear and has been the focus of research. In this review, we summarize the existing research progress, so as to provide guidance for clinical and basic studies. The analgesic mechanism of tuina is mainly manifested in suppressing peripheral inflammation by regulating the TLR4 pathway and miRNA, modulating ion channels (such as P2X3 and piezo), inhibiting the activation of glial cells, and adjusting the brain functional alterations. Overall, tuina has an analgesic effect by acting on different levels of targets, and it is an effective therapy for the treatment of pNP. It is necessary to continue to study the mechanism of tuina analgesia.

Entropy Generation Methodology for Defect Analysis of Electronic and Mechanical Components-A Review.

Understanding the defect characterization of electronic and mechanical components is a crucial step in diagnosing component lifetime. Technologies for determining reliability, such as thermal modeling, cohesion modeling, statistical distribution, and entropy generation analysis, have been developed widely. Defect analysis based on the irreversibility entropy generation methodology is favorable for electronic and mechanical components because the second law of thermodynamics plays a unique role in the analysis of various damage assessment problems encountered in the engineering field. In recent years, numerical and theoretical studies involving entropy generation methodologies have been carried out to predict and diagnose the lifetime of electronic and mechanical components. This work aimed to review previous defect analysis studies that used entropy generation methodologies for electronic and mechanical components. The methodologies are classified into two categories, namely, damage analysis for electronic devices and defect diagnosis for mechanical components. Entropy generation formulations are also divided into two detailed derivations and are summarized and discussed by combining their applications. This work is expected to clarify the relationship among entropy generation methodologies, and benefit the research and development of reliable engineering components.

Photoacoustic-Enabled Self-Guidance in Magnetic-Hyperthermia Fe@Fe3 O4 Nanoparticles for Theranostics In Vivo.

Magnetic nanoparticles have gained much interest for theranostics benefited from their intrinsic integration of imaging and therapeutic abilities. Herein, c(RGDyK) peptide PEGylated Fe@Fe3 O4 nanoparticles (RGD-PEG-MNPs) are developed for photoacoustic (PA)-enabled self-guidance in tumor-targeting magnetic hyperthermia therapy in vivo. In the αv ß3 -positive U87MG glioblastoma xenograft model, the PA signal of RGD-PEG-MNPs reaches its maximum in the tumor at 6 h after intravenous administration. This signal is enhanced by 2.2-folds compared to that of the preinjection and is also 2.2 times higher than that in the blocking group. It demonstrates the excellent targeting property of RGD-PEG-MNPs. With the guidance of the PA, an effective magnetic hyperthermia to tumor is achieved using RGD-PEG-MNPs.

Combination of mesenchymal stem cell injection with icariin for the treatment of diabetes-associated erectile dysfunction.

The present study was aimed to examine whether icariin, a traditional Chinese medicine, could improve therapeutic effects of adipose derived mesenchymal stem cells (ADSCs) for diabetes-associated erectile dysfunction (DMED). DMED were induced in rats by intraperitoneal injection of streptozotocin and confirmed by erectile function measurement. Then, rats of diabetic ED were randomly divided to receive the treatment of saline, ADSCs, icariin or ADSCs combined with icariin respectively. Compared with the treatment by ADSCs or icariin alone, intracavernosum injection of ADSCs combined with the following daily gastric gavage of icariin significantly augmented the value of ICP and ICP/MAP (p<0.01). Meanwhile, the survival of transplanted ADSCs was much improved due to the application of icariin. Similarly, immunofluorescent staining analysis demonstrated that the improved erectile tissue structure by combination of ADSCs and icariin was significantly associated with the increased expression of endothelial markers (vWF) (p<0.01) and smooth muscle markers (α-SMA) (p<0.01). Furthermore, the structure changes in corpus cavernosum were further confirmed by the Masson's trichrome staining. To explore the possible mechanism underlying icariin-enhanced therapeutic efficacy of MSCs, we employed an in vitro testing system by introducing H2O2 to imitate oxidative stress condition considering the oxidative environment faced by engrafted ADSCs and anti-oxidative capacity of icariin. In vitro, we found that the addition of icariin considerably reduced the apoptosis of ADSCs, and attenuated the intracellular reactive oxygen species (ROS), the superoxidase dismutase (SOD) activity and the lactate dehydrogenase (LDH). Subsequently, we examined the expression of apoptosis-related proteins and explored the potential signaling pathway through which icariin promoted the survival of ADSCs against oxidative stress. It was demonstrated that icariin significantly inhibited the upregulation of apoptosis-related proteins under oxidative condition, including Bax and cleaved caspase-3, while promoted the expression of anti-apoptotic factor BCL2. These effects were accompanied with the activation of signal molecules, PI3K/Akt and STAT3. The further signal protein inhibition assays exhibited that the suppression of STAT3 abrogated the icariin-mediated anti-apoptotic effects observed above, while did not influence the expression of PI3K/Akt. However, PI3K inhibition could abrogate icariin-mediated STAT3 activation and achieved a similar effect as STAT3 inhibition. Our results suggested that icariin was an effective adjuvant for enhancing ADSC-based therapy of DMEM, which may be ascribed to their protection of ADSCs against oxidative stress via the regulation of PI3K/Akt-STAT3 signal pathway.

Evaluation of PADUA score as predictor of Warm Ischemia Time (WIT) during Laparoscopic Partial Nephrectomy (LPN).

OBJECTIVE: To evaluate the efficacy of Preoperative Aspects and Dimension Used for an Anatomical (PADUA) classification in the prediction of Warm Ischemia Time (WIT) during Laparoscopic Partial Nephrectomy (LPN). METHODS: Single centre retrospective study on 402 patients who underwent LPN between January 2011 and October 2013. WIT was correlated with each characteristics of the PADUA classification. Univariate and multivariate analysis were used to evaluated the independent predictive risk factors of WIT >20 minutes. RESULTS: The median WIT was 19.8 minutes (IQR 15-25). The PADUA risk groups (High vs Low: OR: 2.804; 95% CI 1.155-6.806; p-value <0.001; Intermediate vs Low: OR 0.971; 95% CI 0.185-5.092 p-value <0.001) and PADUA score as continuous variable (OR: 1.512; 95% CI 1.043-2.194; p-value <0.001) were the best independent risk factors of WIT >20 minutes. Another important factor related with WIT >20 minutes was a CKD stage 3 (p-value: 0.047; OR: 0.528; 95% CI:0.282-0.990). CONCLUSIONS: This study reveal that tumor' anatomical aspects may influence the WIT. The use of the nephrometry scoring systems should to be use by the surgeon in the planning of nephron sparing surgery (NSS) and should be considered for its inclusion in international guidelines.

Efficacy and Safety of Transurethral Photoselective Greenlight(™) Laser Vaporization for the Treatment of Orthotopic Ureteroceles in Adults.

OBJECTIVE: This study aimed to retrospectively evaluate the safety and efficacy of transurethral photoselective Greenlight(™) laser vaporization in adult patients with orthotopic ureterocele. MATERIALS AND METHODS: Thirty adult patients diagnosed with orthotopic urecterocele were recruited at our center. Transurethral photoselective Greenlight laser vaporization was used as the exclusive technique for endoscopic management during the study period. Information, including age, gender, mode of presentation, ureterocele size, vesicoureteral reflux, hydronephrosis status, and incidence of reoperation, were collected for evaluation. RESULTS: Our series included 12 men and 18 women. The mean patient age at presentation was 30.5 years (range, 18-62 years). The mean size of ureterocele was 18 mm (range, 10-41 mm). All patients successfully underwent transurethral photoselective Greenlight laser to vaporize the ureterocele. The operation ranged from 13 min to 38 min (mean 19.6 min). The average blood loss was <10 mL. No patient had intraoperative complications. The average postoperative hospital stay was 18.3 h. All patients were voided after postoperative catheter removal. None of the patients demonstrated any residual ureterocele and/or hydronephrosis when evaluated with ultrasonography after 3 months. Only one patient with a duplex collecting system presented asymptomatic low-grade reflux at 3 months, which was spontaneously resolved after 6 months of follow-up. All patients were free of any symptoms. No reoperative procedures were required at a mean follow-up of 14.2 months (range, 8-16). CONCLUSIONS: Transurethral photoselective Greenlight laser vaporization is safe, effective, and efficient for the management of orthotopic urecteroceles in adults. Therefore, this technique should be considered as the initial treatment in most patients.

Evaluation of laparoscopic vs robotic partial nephrectomy using the margin, ischemia and complications score system: a retrospective single center analysis.

OBJECTIVE: To evaluate differences between Laparoscopic Partial Nephrectomy (LPN) and Robot-Assisted Partial Nephrectomy (RAPN) using the Margin, Ischemia and Complications (MIC) score system and to evaluate factors related with MIC success. MATERIALS AND METHODS: Single centre retrospective study on 258 LPN and 58 RAPN performed between January 2012 and January 2014. Success was defined when surgical margins was negative, Warm Ischemia Time (WIT) was ≤ 20 minutes and no major complications occurred. Mann-Whitney-U and Pearson χ2 correlation were used to compare LPN and RAPN. A matched pair comparison was also performed. Spearman correlation (Rho) was used to evaluate the relationship between clinical, intra and post-operative and pathological patients characteristics with MIC score. A binary regression analysis was also performed to evaluate independent factors associated with MIC success. RESULTS: The MIC rate in LPN and RAPN was 55% and 65.5% respectively. No differences in clinical, intra and post-operative outcomes between groups were found. Clinical tumor size (p-value: < 0.001; OR: 0.829; 95% CI: 0.697-0.987), PADUA score (p-value: < 0.001; OR: 0.843; 95% CI: 0.740-0.960), PADUA risk groups (intermediate; p-value: < 0.001; OR: 0.416; 95% CI: 0.238- 0.792; high: p-value: < 0.001; OR: 0.356; 95% CI: 0.199- 0.636), WIT (p-value: < 0.001; OR: 0.598; 95% CI: 0.530- 0.675) were independently associated with MIC. eGFR (< 60 vs ≥ 60 ml/min per 1.73 m2: p-value: < 0.001; OR: 3.356; 95% CI: 1.701-6.621) and Fuhrman nuclear grade (p-value: 0.014; OR: 1.798; 95% CI:1.129-2.865) were also independently associated with MIC. CONCLUSIONS: MIC score system is a simple and useful tool to report and to compare different surgical approach.

Hypoxia precondition promotes adipose-derived mesenchymal stem cells based repair of diabetic erectile dysfunction via augmenting angiogenesis and neuroprotection.

The aim of the present study was to examine whether hypoxia preconditioning could improve therapeutic effects of adipose derived mesenchymal stem cells (AMSCs) for diabetes induced erectile dysfunction (DED). AMSCs were pretreated with normoxia (20% O2, N-AMSCs) or sub-lethal hypoxia (1% O2, H-AMSCs). The hypoxia exposure up-regulated the expression of several angiogenesis and neuroprotection related cytokines in AMSCs, including vascular endothelial growth factor (VEGF) and its receptor FIK-1, angiotensin (Ang-1), basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF), stromal derived factor-1 (SDF-1) and its CXC chemokine receptor 4 (CXCR4). DED rats were induced via intraperitoneal injection of streptozotocin (60 mg/kg) and were randomly divided into three groups-Saline group: intracavernous injection with phosphate buffer saline; N-AMSCs group: N-AMSCs injection; H-AMSCs group: H-AMSCs injection. Ten rats without any treatment were used as normal control. Four weeks after injection, the mean arterial pressure (MAP) and intracavernosal pressure (ICP) were measured. The contents of endothelial, smooth muscle, dorsal nerve in cavernoursal tissue were assessed. Compared with N-AMSCs and saline, intracavernosum injection of H-AMSCs significantly raised ICP and ICP/MAP (p<0.05). Immunofluorescent staining analysis demonstrated that improved erectile function by MSCs was significantly associated with increased expression of endothelial markers (CD31 and vWF) (p<0.01) and smooth muscle markers (α-SMA) (p<0.01). Meanwhile, the expression of nNOS was also significantly higher in rats receiving H-AMSCs injection than those receiving N-AMSCs or saline injection. The results suggested that hypoxic preconditioning of MSCs was an effective approach to enhance their therapeutic effect for DED, which may be due to their augmented angiogenesis and neuroprotection.

Effects of continuous passage on immunomodulatory properties of human adipose-derived stem cells.

Human adipose-derived stem cells (hADSCs) have the ability to influence immune response, and hence are key cell sources for tissue repair and regeneration. In this study we explored the effect of continuous passage on the immunomodulatory properties of hADSCs to provide some advises for large-scale production of hADSCs for clinical applications. We found that after continuous passage, the specific surface markers expression levels as well as the adipogenic and osteogenic differentiation capacities of hADSCs had no obvious changes. However, the secretion levels of IL-10 and HGF reduced dramatically along with passage numbers. Furthermore, the INF-γ level was found higher in which medium peripheral blood mononuclear cells were co-cultured with hADSCs with higher passage numbers. Also, the in vivo experiments showed that the peritonitis model mice, which were injected with higher passage numbers of hADSCs, tended to have higher levels of inflammation. All these together indicated that continuous passage has only minor effect on the cell phenotypes but will impair the immunomodulatory properties of hADSCs. This suggests that hADSCs could be prepared by continuous passage, but only those cells of lower passage numbers would be ideal therapeutic tools.

[Suppression of tumor growth in renal cancer treatment with tumor vaccination after haploidentical bone marrow cell reconstitution].

OBJECTIVE: To investigate whether whole tumor cell vaccination strategies in combination with bone marrow transplantation (BMT) can stimulate graft-versus-tumor effect (GVT). METHODS: Twenty-six BALB/c mice were randomly divided into 3 groups: BMT group (group A, n = 10), BMT + vaccination group (group B, n = 10), control group (group C, n = 6). (BALB/c × C57BL/6) F1 mice [CB6F1, H-2K(b/d)] were used as donors. BALB/c mice of group C were only inoculated with Renca cell (2.6 × 10(6)). Mice of group A and B were conditioned with 8 Gy irradiation, followed by infusion by bone marrow cell of CB6F1 mice on day 1, then inoculated with Renca cell (2.6 × 10(6)) on day 8. All mice of group B were immunized subcutaneous on the back with 5 × 10(5) irradiated Renca tumor cells on day 9 and day 16. All mice of group C were inoculated with Renca cell (2.6 × 10(6)) on day 8. In group A and B, all mice were analyzed by fluorescence activated cell sorter (FACS) on day 14, and 28 day after BMT. Mice were killed on day 32 after inoculation with tumor cell and collected blood sample. All tumors were taken out to be weighed and then fixed in 10% buffered formalin, embedded in paraffin, and cut into 5 µm slices. The slices were stained with HE and examined by TdT mediated-dUTP nick end labeling (TUNEL). Liver, skin, intestine, and spleen were biopsied for histopathological examination. RESULTS: The results of chimera showed that engraftments of group A, B were full donor chimerism, and the chimerism of those remained above 90% and preserved even after 28 days. The tumor weight, tumor volume increment in the group B was lower than group A and C (P < 0.05). The tumor suppressing rates of the group A and B were 54%, 60% respectively. The area ratio of tumor necrosis and apoptosis index (AI) of the tumor in the group B were higher than group A and C (P < 0.05). Graft-versus-host disease was not observed in each group. CONCLUSION: The mechanism of GVT after haploidentical allogeneic bone marrow transplantation with tumor vaccination may be the promotion of tumor necrosis and apoptosis.

[Apoptosis and expression of caspase-3 protein in germ cells of varicocele rats].

OBJECTIVE: To explore the relationship between the expression of caspase-3 in testicular germ cells of rats with experimental left varicocele (ELV) and apoptosis of germ cells. METHODS: Twenty-four male SD rats were randomly divided into three groups with eight animals each: sham-operation group (SOG), 30-day post-operation group (PG1) and 60-day psot-operation group (PG2). ELV model was established by the partial ligation of the left renal vein. To detect apoptosis of germ cells and expression of caspase-3, TUNEL assay and immunohistochemistry (SABC) were used respectively. RESULTS: The number of caspase-3 positive germ cells per tubular cross section in left and right testes of rats in SOG, PG1, PG2 were 0.1175 +/- 0.0129, 0.2463 +/- 0.0421, 0.2938 +/- 0.0511 and 0.1650 +/- 0.0192, 0.2538 +/- 0.0219, 0.2775 +/- 0.0343, respectively. Compared with SOG, the expression of caspase-3 in bilateral testes of rats in PG1 and PG2 were increased, and the differences were statistically significant(P = 0.0115 and P = 0.0144). CONCLUSION: Expression of caspase-3 protein increased in germ cells of rats with ELV, which may be one of the molecular mechanisms related to excessive testicular germ cell apoptosis.