A GP130-Targeting Small Molecule, LMT-28, Reduces LPS-Induced Bone Resorption around Implants in Diabetic Models by Inhibiting IL-6/GP130/JAK2/STAT3 Signaling.

In this study, we examined the effect of the GP130-targeting molecule, LMT-28, on lipopolysaccharide- (LPS-) induced bone resorption around implants in diabetic models using in vitro and rat animal experiments. First, LMT-28 was added to osteoblasts stimulated by LPS and advanced glycation end products (AGEs), and nuclear factor-κB receptor-activating factor ligand (RANKL) and associated pathways were evaluated. Then, LMT-28 was administered by gavage at 0.23 mg/kg once every 5 days for 2 weeks to type 2 diabetic rats with peri-implantitis induced by LPS injection and silk ligature. The expression of IL-6 and RANKL was evaluated by immunohistochemistry, and the bone resorption around implants was evaluated by microcomputed tomography. The results showed that LMT-28 downregulated the expression of RANKL through the JAK2/STAT3 signaling pathway in osteoblasts stimulated by LPS and AGEs, reduced bone resorption around implants with peri-implantitis, decreased the expression of IL-6 and RANKL, and decreased osteoclast activity in type 2 diabetic rats. This study confirmed the ability of LMT-28 to reduce LPS-induced bone resorption around implants in diabetic rats.

Optimization and performance evaluation of double-stranded probe in real-time PCR.

BACKGROUND: TaqMan probe-based real-time PCR (qPCR/RT-qPCR) has been widely used in various fields because of its high sensitivity and specificity. However, TaqMan probes are associated with a relatively higher background signal, and hence negatively affect the detection results. METHODS: Double-stranded probes (DSPs) were designed for the high sensitive detection of hepatitis B virus (HBV) DNA and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA using qPCR/RT-qPCR. Old DSPs (ODSPs) consist of different lengths of positive and negative strands with complementary oligonucleotides. We systematically optimized ODSPs length, the free energy of hybridization (ΔG) between complementary oligonucleotides, and the length of sticky ends, and the novel DSPs performances were evaluated in comparison with other types of probes. RESULTS: By using similar length positive and negative strands, controlling ΔG between complementary oligonucleotides to approximately -30 kcal/mol, and maintaining the sticky end length at 4-6 nt, the analytical performances of DSPs were significantly improved. Compared with other types of probes, DSPs are advantageous in fluorescence signal intensity and sensitivity. CONCLUSION: DSPs can further improve the detection sensitivity and the detection rate of low-concentration samples in molecular diagnosis.

Evaluation of longitudinal trajectory of functional tricuspid regurgitation on the risk of right ventricular dysfunction after mitral valve replacement.

OBJECTIVE: Functional tricuspid regurgitation (FTR) levels can vary over time and its longitudinal changing patterns may predict right ventricular dysfunction (RVD) risk. We aim to identify different trajectories of FTR in those who received mitral valve replacement (MVR) and investigate the association between longitudinal trajectory groups and RVD risk in a cohort study. METHODS AND RESULTS: A prospective cohort study, reported usual FTR levels at baseline in 2005-2015 and the participants of MVR have been followed up for 5-6 years, approximately every 1 year, and so far, the data have been collected across five subsequent phases. Five-year longitudinal trajectories of FTR were identified using group-based trajectory modeling (GBTM). We identified 3 distinct trajectories using a GBTM, labeled by initial value and changing pattern: stable group (258/378, 68.2%), increasing-slow group (67/378, 17.6%) and increasing-fast group (53/378, 14.2%). Treating the stable group as the reference, the age- and sex-adjusted odds ratio (OR) was 25.84 (95% confidence interval [CI]: 11.78-56.65) for the increasing-slow group and 139.94 (95% CI: 45.47-430.68) for the increasing-fast group by logistic regression model. After adjustment for every potential confounding factors, the OR is 14.21 (95% CI: 4.36-46.33) and 49.34 (95% CI: 8.88-273.87), respectively. CONCLUSIONS: The longitudinal trajectories of worsening FTR were mostly associated with increased risk of RVD outcomes, which is independent of other factors including FTR levels. These findings have implications for intervention and prevention of RVD among individuals who received MVR.

Establishment and Validation of a Predictive Model for Long-Term Severe Functional Tricuspid Regurgitation after Mitral Valve Replacement.

BACKGROUND: The objective was to develop and validate an individualized nomogram to predict severe functional tricuspid regurgitation (S-FTR) after mitral valve replacement (MVR) via retrospective analysis of rheumatic heart disease (RHD) patients' pre-clinical characteristics. METHODS: Between 2001-2015, 442 MVR patients of RHD were examined. Transthoracic echocardiography detected S-FTR, and logistic regression model analyzed its independent predictors. R software established a nomogram prediction model, and Bootstrap determined its theoretical probability, which subsequently was compared with the actual patient probability to calculate the area under the curve (AUC) and calibration plots. Decision curve analysis (DCA) identified its clinical utility. RESULTS: Ninety-six patients developed S-FTR during the follow-up period. Both uni- and multivariate analyses found significant correlations between S-FTR occurrence with gender, age, atrial fibrillation (AF), pulmonary arterial hypertension (PH), left atrial diameter (LAD), and tricuspid regurgitation area (TRA). The individualized nomogram model had the AUC of 0.99 in internal verification. Calibration test indicated high agreement of predicted and actual S-FTR onset. DCA also showed that utilization of those six aforementioned factors was clinically useful. CONCLUSION: The nomogram for the patient characteristics of age, gender, AF, PH, LAD, and TRA found that they were highly predictive for future S-FTR onset within 5 years. This predictive ability therefore allows clinicians to optimize postoperative patient care and avoid unnecessary tricuspid valve surgeries.

[Repair impact of vibration exercise with different frequencies on articular cartilage of rats with early knee osteoarthritis and its JNK/NF-κB, SOX9 mechanisms].

Objective: To investigate the repair effect and JNK/NF-κB,SOX9 mechanisms of vibration exercise with different frequencies on articular cartilage in rats with early knee osteoarthritis. Methods: Forty-eight adult male SD rats were randomly divided into six groups(n=8):model control group(MC),high frequency vibration group 1 (GP1,60 Hz),high frequency vibration 2 group (GP2,40 Hz),medium frequency vibration group (ZP,20 Hz),minor frequency group(DP,10 Hz)and normal control group(NC). Except for NC group,the rats in each group were made into early knee osteoarthritis model after six weeks of knee joint cavity injection of papain solution and 2% mixture l-cysteine on the 1st,4 th and 7th day. Each exercise group was subjected vibration to 40 minutes a day with amplitude of 2～5 mm and 5 days a week. Four weeks later, the articular cartilage of the lateral femoral condyle of the both back leg knee joints were detected by HE staining,serine O staining and Mankin scores for morphological observation. The expression levels of JNK,NF-κB p65 and Sox9 mRNA in articular cartilage of the medial femoral condyle were detected by RT-qPCR,and the protein expressions of JNK,NF-κB p65 and Sox9 were detected by Western blot. Results: Compared with the NC group,the Mankin score in other groups was significantly higher (P＜0.01). Compared with the MC group,the Mankin score of each vibration group was significantly lower(P＜0.05),the mRNA and protein expressions of JNK and NF-κB p65 in each vibration training group were significantly lower (P＜0.01),the expressions of Sox9 mRNA and protein in vibration training group were increased significantly (P＜0.01). Compared with the higher frequency group,the Mankin score,the mRNA and protein expressions of JNK and NF-κB p65 of lower frequency group were significantly lower (P＜0.05 or P＜0.01). But the expressions of Sox9 mRNA and protein were significantly higher (P＜ 0.05 or P＜0.01). Conclusion: Vibration exercise of different frequencies may present varying degrees of cartilage repair impact in rats with early knee osteoarthritis,and the cartilage repair by low-frequency vibration training is better than that by high-frequency vibration. This can be one of the mechanisms on controlling collagen synthesis by down-regulating JNK/NF-κB expression and increasing SOX9 activity of OA articular cartilage.

Left atrial strain predicts risk and prognosis in patients with acute coronary syndrome: A retrospective study with external validation.

Objectives: To explore the association between left atrial (LA) strain and the GRACE score in patients with acute coronary syndrome (ACS) and to investigate the utility of LA function in predicting short-term adverse cardiovascular events post ACS. Methods: This retrospective study included ACS patients who underwent coronary angiography (CAG) in two independent cohorts from October 2020 to July 2022. The patients were classified into low-intermediate risk group and high-risk group based on the GRACE score. All participants underwent a transthoracic echocardiogram, with LA strain analysis before CAG. Correlation analysis was used to determine the relationship between LA strain and the GRACE score. The predictive value of LA strain was examined utilizing the area under the curve (AUC). Participants were followed for 10.5 ± 2.9 months for the primary endpoint of major adverse cardiovascular events (MACE). Results: A total of 229 patients were included in this study, including 196 in the primary group and 33 in the validation group. Spearman's correlation analysis showed there was a moderate negative correlation between the GRACE and left atrial reservoir strain (LASr) in both the primary (r = -0.63, P < 0.001) and validation (r = -0.73, P < 0.001) cohorts. Receiver operator characteristic (ROC) curve analysis showed that the AUC of LASr for prediction of the high-risk group was 0.86. Taking LASr 19.6% as the cut-off value, the sensitivity and specificity were 0.71 and 0.92, respectively. The cut-off value of 19.6% remains good at identifying high-risk group in the validation group (AUC = 0.87, sensitivity: 77.8%, specificity: 95.8%). Furthermore, 49 patients reached the endpoint in the primary cohort during the follow-up. On multivariable regression analysis, LASr (P = 0.03) was the independent echocardiographic predictor for the primary endpoint, rather than left atrial volume index (LAVI). Conclusions: LASr can identify high-risk patients with ACS as defined by the GRACE score and may be superior to Max LAVI in predicting incidents of MACE in the short-term following ACS.

Enhanced Separation Efficiency and Purity of Circulating Tumor Cells Based on the Combined Effects of Double Sheath Fluids and Inertial Focusing.

Circulating tumor cells (CTCs) play a crucial role in solid tumor metastasis, but obtaining high purity and viability CTCs is a challenging task due to their rarity. Although various works using spiral microchannels to isolate CTCs have been reported, the sorting purity of CTCs has not been significantly improved. Herein, we developed a novel double spiral microchannel for efficient separation and enrichment of intact and high-purity CTCs based on the combined effects of two-stage inertial focusing and particle deflection. Particle deflection relies on the second sheath to produce a deflection of the focused sample flow segment at the end of the first-stage microchannel, allowing larger particles to remain focused and entered the second-stage microchannel while smaller particles moved into the first waste channel. The deflection of the focused sample flow segment was visualized. Testing by a binary mixture of 10.4 and 16.5 µm fluorescent microspheres, it showed 16.5 µm with separation efficiency of 98% and purity of 90% under the second sheath flow rate of 700 µl min-1. In biological experiments, the average purity of spiked CTCs was 74% at a high throughput of 1.5 × 108 cells min-1, and the recovery was more than 91%. Compared to the control group, the viability of separated cells was 99%. Finally, we validated the performance of the double spiral microchannel using clinical cancer blood samples. CTCs with a concentration of 2-28 counts ml-1 were separated from all 12 patients' peripheral blood. Thus, our device could be a robust and label-free liquid biopsy platform in inertial microfluidics for successful application in clinical trials.

A comparative study on the use of palm bark as a supplementary carbon source in partially saturated vertical constructed wetland: Organic matter characterization, release-adsorption kinetics, and pilot-scale performance.

In this study, a comparative investigation of palm bark and corncob (a well-investigated material) for enhancing nitrogen removal efficiency in partially saturated vertical constructed wetland (PSVCW) was performed to evaluate an effective and cost-effective supplementary carbon source. The characteristics of the released organic matter and the release processes were analyzed through optical property characterization and a first-order release-adsorption model, respectively, and the nitrogen removal performance was evaluated in a series of pilot-scale PSVCWs. Results showed that the amount of organic matter released per unit mass of corncob was larger than that released per unit mass of palm bark under the same pretreatment conditions (control, heat, and alkaline pretreatment). The organic matter released from corncob has a higher apparent molecular weight and a higher degree of aromatic condensation than those of the organic matter released from palm bark, whereas the organic matter released from palm bark has higher and more stable bioavailability. Moreover, palm bark showed a more significant improvement of release capacity with the heat and alkaline pretreatment methods. Pilot-scale studies revealed that PSVCW using palm bark as the supplementary carbon source has a longer replacement cycle and higher total nitrogen (TN) removal efficiency than that using corncob, indicating that palm bark can be considered an effective and inexpensive supplementary carbon source. This study provides initial guidance for the ongoing research on supplementary carbon sources for improving nitrogen removal efficiency in constructed wetlands.

[Effect of electronic moxibustion on memory function in patients with amnestic mild cognitive impairment].

OBJECTIVE: To observe the effect of electronic moxibustion on memory function in the patients with amnestic mild cognitive impairment (aMCI). METHODS: A total of 59 aMCI patients were randomized into an electronic moxibustion group (30 cases) and a placebo moxibustion group (29 cases). In the electronic moxibustion group, the electronic moxibustion was applied to Baihui (GV 20), Dazhui (GV 14), Mingmen (GV 4) and Taixi (KI 3), 45 ℃ in temperature, 20 min each time. The treatment was given once a day, 5 times a week. The treatment for 4 weeks was as one course and 2 courses were required totally. In the placebo moxibustion group, the moxa-free patch was used, 38 ℃ in temperature. The acupoint selection and the treatment frequency were same as the electronic moxibustion group. Before and after treatment, Rivermead behavior memory test (RBMT) was adopted to evaluate the global memory function of the patients in the two groups and the N-back task test was adopted to evaluate working memory function separately. Additionally, the mini-mental state examination (MMSE) and its immediate memory, Montreal cognitive assessment (MoCA) and its delay recall were adopted to evaluate the global cognitive function and memory function. RESULTS: In the electronic moxibustion group, after treatment, RBMT score, N-back accuracy rates, MMSE and MoCA scores and the scores of immediate memory and delay recall were improved significantly as compared with those before treatment (P<0.01). In the placebo moxibustion group, the accuracy rates of 1-back and 2-back task and the scores of immediate memory and delay recall were improved obviously as compared with those before treatment (P<0.05, P<0.01). After treatment, the improvements of RBMT score, the accuracy rates of N-back task and MMSE and MoCA scores in the electronic moxibustion group were higher than those in the placebo moxibustion group (P<0.05). CONCLUSION: Electronic moxibustion improves memory function in the patients with amnestic mild cognitive impairment.

A Systems Pharmacology Approach for Identifying the Multiple Mechanisms of Action for the Rougui-Fuzi Herb Pair in the Treatment of Cardiocerebral Vascular Diseases.

Cardiocerebral vascular diseases (CCVDs) are the main reasons for high morbidity and mortality all over the world, including atherosclerosis, hypertension, myocardial infarction, stroke, and so on. Chinese herbs pair of the Cinnamomum cassia Presl (Chinese name, rougui) and the Aconitum carmichaelii Debx (Chinese name, fuzi) can be effective in CCVDs, which is recorded in the ancient classic book Shennong Bencao Jing, Mingyibielu and Thousand Golden Prescriptions. However, the active ingredients and the molecular mechanisms of rougui-fuzi in treatment of CCVDs are still unclear. This study was designed to apply a system pharmacology approach to reveal the molecular mechanisms of the rougui-fuzi anti-CCVDs. The 163 candidate compounds were retrieved from Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP). And 84 potential active compounds and the corresponding 42 targets were obtained from systematic model. The underlying mechanisms of the therapeutic effect for rougui-fuzi were investigated with gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Then, component-target-disease (C-T-D) and target-pathway (T-P) networks were constructed to further dissect the core pathways, potential targets, and active compounds in treatment of CCVDs for rougui-fuzi. We also constituted protein-protein in interaction (PPI) network by the reflect target protein of the crucial pathways against CCVDs. As a result, 21 key compounds, 8 key targets, and 3 key pathways were obtained for rougui-fuzi. Afterwards, molecular docking was performed to validate the reliability of the interactions between some compounds and their corresponding targets. Finally, UPLC-Q-Exactive-MSE and GC-MS/MS were analyzed to detect the active ingredients of rougui-fuzi. Our results may provide a new approach to clarify the molecular mechanisms of Chinese herb pair in treatment with CCVDs at a systematic level.

[Professor ZHANG Qing-ping's experience in the treatment of vascular dementia with acupuncture and moxibustion].

In the aspects of the acupoint prescription, acupuncture techniques and typical cases, the characteristics and experience of professor ZHANG Qing-ping in the clinical treatment of vascular dementia with acupuncture and moxibustion were introduced. Vascular dementia is treated on the base of etiology, focusing on the brain as the root cause, with the reinforcing and the promoting as the dominant principle of acupuncture. The treatment is emphasized on tonifying the kidney to fill up marrow, strengthening the spleen to nourish the brain and promoting the collateral circulation to improve the intelligence. Concerning to the general situation of the disease, regulating mind stressed in treatment. In clinical practice, Baihui (GV 20) is the chief acupoint to benefit the brain. Yuan-source points and the eight influential points are predominated to tonify the functions of zangfu organs and fill up vessels. Moxibustion is adopted to resolve the stagnation and promote circulation in meridian as well as regulate qi and blood circulation. Specially, the moxibustion technique for resolving the stagnation and promoting circulation in meridian achieves the satisfactory therapeutic effects in clinical treatment.

Survival following radiotherapy in young women with localized early-stage breast cancer according to molecular subtypes.

BACKGROUND: To evaluate the significance and benefit of radiotherapy (RT) in young early-stage breast cancer patients according to different molecular subtypes. METHODS: We conducted a retrospective cohort study utilizing the Surveillance, Epidemiology, and End Results database with known hormone receptor (HoR) and human epidermal growth factor receptor 2 (HER2) status. Female patients aged 18-45, received RT treatment, and diagnosed with stage T1-3, N0-3, M0 primary breast cancer between 2010 and 2013 were identified. RESULTS: Of all the 23 148 included patients, 14 708 (63.54%), 3385 (14.62%), 1225 (5.29%), and 3830 (16.55%) were diagnosed with luminal-A (HoR + HER2-), luminal-B (HoR + HER2+), HER2-enriched (HoR-HER2+), and triple-negative (HoR-HER2-) breast cancer, respectively. RT was significantly correlated with improved overall survival (OS, HR: 0.295; 95% CI:0.138-0.63, P = 0.002) and breast cancer-specific survival (BCSS, HR: 0.328; 95% CI: 0.153-0.702, P = 0.004) in HER2-enriched patients. In addition, a significantly prolonged OS was also observed when RT was given to luminal-A (HR: 0.696; 95% CI: 0.538-0.902, P = 0.006) and luminal-B (HR: 0.385; 95% CI:0.199-0.744, P = 0.005) breast cancer patients compared to those without RT. Multivariable-adjusted analyses showed that HER2 was a significant favorable factor for RT benefit in breast cancer patients. CONCLUSIONS: RT could offer significant survival benefit in luminal-A, luminal-B, and especially HER2-enriched young early-stage breast cancer female patients. The results enabled clinicians to predict the benefits of RT and improve evidence-based treatment for breast cancer patients.