[Application of cardiac magnetic resonance imaging in subclinical systemic sclerosis primary heart involvement].

Objectives: Primary cardiac involvement (SSc-PHI) in systemic sclerosis is an important prognostic factor. We aimed to characterize and identify subclinical SSc-PHI using cardiovascular MRI to determine whether disease severity and serum biomarkers are associated with subclinical SSc-PHI. Methods: A total of 26 patients with SSc who had no history of cardiovascular disease or pulmonary hypertension underwent 3 T-enhanced cardiovascular MRI. Measurements included native T1, extracellular volume, advanced gadolinium enhancement, T2 mapping, and left ventricular volume function. Troponin T and N telencephalic natriuretic peptide precursors were also determined. Results: LGE was observed in 13 of 26 patients (50.0%), suggesting focal fibrosis, and T2 mapping was significantly higher in the dcSSc group than in the lcSSc group (P=0.009). Left ventricular volume and function were within the normal range in all patients, but final systolic left ventricular volume was significantly higher in dcSSc than in lcSSc (P=0.021). The modified Rodnan skin score (mRSS) was significantly higher in patients with LGE focal fibrosis (P=0.019). Logistic regression analysis confirmed the association between mRSS and LGE (OR=1.224, P=0.037). In multivariate analysis, T2 mapping was negatively correlated with disease course, and was correlated with dcSSc and fingertip ulcer (R2=0.711, P=0.018, P=0.013, P=0.030). Troponin T was correlated with T2 mapping (r=0.555, P=0.049). Conclusions: Subclinical SSc-PHI is characterized by diffuse and focal myocardial fibrosis, but preserves myocardial systolic function. Subclinical SSC-Phi is associated with TNT, SSc disease severity, and complex peripheral vascular disease. These data provide information for identifying individuals at risk of SSc-PHI.

[Distribution characteristics of tumor infiltrating lymphocytes in EBV-associated lymphoepithelioma-like carcinoma and their clinical significance].

Objective: To investigate the association between the distribution of tumor infiltrating lymphocytes (TIL) in EBV associated lymphoepitheliomatoid carcinoma (LELC) and the pathological subtypes of LELC, as well as the clinical significance of TIL distribution. Methods: The LELC patients with sufficient tumor tissues, complete clinical data and positive EBER, who visited Zhejiang Cancer Hospital, Hangzhou, China from January 2006 to October 2018, were selected. Various immunohistochemical markers (CD20, CD138, CD4, CD8, CD56 and FOXP3) were examined for TIL typing. Two pathologists reviewed the hematoxylin and eosin (HE) staining sections and interpreted the immunohistochemical results. Correlation analysis was used to evaluate the relationship between the distribution of TIL subgroups and LELC's pathological characteristics. Survival analyses were conducted to study the prognostic values of TIL subgrouping. Results: A total of 102 patients with EBV related LELC were included. 46 of them were classic LELC (c-LELC) with rich interstitial TIL, and 56 were non-classic LELC (n-LELC) with relatively fewer interstitial TIL. The results of TIL analysis showed that all subtypes of c-LELC were rich in TIL, with B lymphocytes as the dominant subgroup. The number of TIL in n-LELC was fewer than that in c-LELC, with T lymphocytes as the dominant subgroup. There was no significant difference in the distribution of plasma cells between the two groups. Survival analysis showed that the total number of TIL, and the infiltrations of CD20+B cells, CD4+T cells, and FOXP3+Treg cells were associated with better overall survivals (P=0.004, 0.003, 0.008 and 0.025, respectively) and disease-free survivals (P=0.011, 0.003, 0.038 and 0.041, respectively) in patients with LELC. Conclusions: The morphologic subtypes of EBV-related LELC have different tumor immune characteristics. The total number of TIL in the stroma of c-LELC is significantly higher than that of n-LELC. Interestingly, B lymphocytes are the dominant TIL in c-LELC, while T lymphocytes are the dominant TIL in n-LELC. The infiltration of TIL, CD20+B cells, CD4+T cells and FOXP3+Treg cells in LELC may suggest a better prognosis.

[The incidence and causes of Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia].

Objective: To observe the incidence of Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia and to explore the risk factors for its occurrence. Methods: The data of patients suffered from Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia from June 2018 to May 2021 were retrospectively reviewed. The type of procedure, surgeon, age and gender were selected as matching factors, 4 patients without Tapia syndrome were selected as control group for each case. The radiological parameters including mandibular-vertebral distance, thyroid-vertebral distance, thyroid cartilage-vertebral distance, and C2-C7 lordotic Cobb angle were measured on lateral radiographs of the cervical spine. The above parameters were measured on neutral, over-flexion and over-extension radiographs. The difference between the Tapia group and the control group were analyzed. Results: There were 9 patients (0.37%) suffered from Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia in 2 431 patients, and it happened in 0.67 days (0-2 days) after the operation. There were 3 males and 6 females with a mean age of (61±5) years. The clinical manifestations was tongue extension deviation in 8 cases (88.9%), dysarthria in 6 cases (66.7%), dysphagia in 3 cases (33.3%), tongue stiffness in 3 cases (33.3%), hoarseness in voice and pharyngeal discomfort in 1 case (11.1%). All of the symptoms were relieved in all patients at 3 months postoperative follow-up. In neutral position, the mandibular-vertebral distance was (7.19±3.96) mm in the control group and it was (3.98±3.01) mm in Tapia group (P<0.05). From neutral position to hyperflexion position, the distance between mandible and vertebral body was reduced from 3.98 mm to 1.95 mm in the Tapia group and decreased for 51.0%, and it decreased from 7.19 mm for 31.8% to 4.90 mm in the control group. Conclusions: The incidence of Tapia syndrome after posterior cervical spine surgery under oral tracheal intubation general anesthesia is low. A smaller mandibular-vertebral distance on pre-operative cervical spine lateral view radiograph maybe a risk factor for Tapia syndrome after posterior cervical surgery under oral tracheal intubation general anesthesia.

[Efficacy comparison of minimally invasive atlantoaxial mass fusion and open atlantoaxial fusion in the treatment of atlantoaxial dislocation].

Objective: To compare the clinical effects of minimally invasive intermuscular atlantoaxial lateral mass fusion (Mis-PALF) and open atlantoaxial fusion in patients with atlantoaxial dislocation. Methods: The clinical data of patients with atlantoaxial dislocation who received Mis-PALF operation (17 cases) or open atlantoaxial fusion (88 cases, as control) in the Third Hospital of Peking University from September 2015 to September 2021 were analyzed retrospectively. In Mis-PALF group, there were 9 males and 8 females, aged (45.8±19.8) years; and there were 48 males and 40 females in the control group, aged (50.0±13.9) years. The operation time, perioperative blood loss, postoperative body temperature, postoperative pain [assessed with visual analogue scale (VAS)], postoperative additional analgesic drugs, postoperative hospitalization time, the improvement rate of Japanese Orthopedic Association (JOA) scores of spinal cord function in three-months follow-up and complications were compared between the two groups. Results: Mis-PALF group had less perioperative blood loss than control group [(111.8±35.9)ml vs (362.9±18.6)ml, P<0.01], shorter hospitalization time [(3.06±0.63) days vs (4.24±0.14) days, P<0.01] and fewer additional analgesic drugs (3/17 vs 56/88, P<0.01). There was no significant difference between the Mis-PALF and control group in operation time [(125±7)min vs (113±8)min, P=0.525], patients with fever(11/17 vs 37/88, P=0.086) or postoperative pain (1/17 vs 13/88, P=0.357), the improvement rate of JOA score (66.9%±28.8% vs 74.2%±28.6%, P=0.409) and complications rate (1/17 vs 3/88, P=1.000). Conclusion: Mis-PALF can significantly reduce the perioperative blood loss, shorten the postoperative hospitalization time and the additionally use of analgesic drugs without increasing complications.

[Application progress of liquid chromatography tandem mass spectrometry in precision medicine].

Liquid chromatography tandem mass spectrometry (LC-MS/MS) is an analytical method that combines high separation of liquid chromatography with high selectivity and sensitivity of mass spectrometry. In recent years, LC-MS/MS has been widely used in clinical practice, including screening of inherited disorders, determination of endogenous compounds and analysis of biomarkers. LC-MS/MS is of great value to the precision prevention, diagnosis and treatment of some diseases due to its accurate data. This article not only illustrates the advantages of LC-MS/MS in precision medicine, but also prospects the future trend of LC-MS/MS in clinical practice, which expects to promote the development of clinical LC-MS/MS in the prevention, diagnosis and treatment of diseases.

[Primary ovarian squamous cell carcinoma: clinicopathological features and prognostic analysis of fifteen cases].

Objective: To assess the clinical features and treatment outcomes in patients with primary ovarian squamous cell carcinoma (POSCC). Methods: Fifteen patients with primary ovarian squamous cell carcinoma diagnosed from January 2009 to December 2018 in Cancer Hospital of the University of Chinese Academy of Sciences were collected. The expression of p16, hMLH1, hMSH2, hMSH6 and PMS2 in POSCC was detected by immunohistochemistry, and the status of high-risk human papillomavirus (HPV) by RNAscope test. Results: Squamous cell carcinoma with different degrees of differentiation was found in 15 cases, including three cases with high differentiation and 12 cases with medium to low differentiation. There were four cases with in situ squamous cell carcinoma, four cases with teratoma, one case with endometrial carcinoma/atypical hyperplasia, and one case with endometriosis. p16 was expressed in five cases (5/15), indicating coexisting high-risk HPV infection. There was no high-risk HPV infection in the remaining 10 cases, and p16 staining was negative. There was no deficient mismatch repair protein in all cases. The overall survival time (P=0.038) and progression free survival (P=0.045) of patients with high-risk HPV infection were longer than those without HPV infection. Conclusions: POSCC is more commonly noted in postmenopausal women and often occurs unilaterally. Elevated serological indexes CA125 and SCC are the most common finding. Morphologically, the tumors show variable degrees of differentiation, but the current data suggest that the degree of differentiation cannot be used as an independent prognostic index. High-risk HPV infection may be associated with the occurrence of POSCC, and that the prognosis of POSCC patients with HPV infection is better than that of patients without infection.

[Relationship between onco-immunological and morphologic characteristics of lymphoepithelioma-like carcinoma and lymphocyte subtypes of peripheral blood].

Objective: To study the relationship between the onco-immunological and morphologic characteristics of lymphoepithelioma-like carcinoma (LELC) and peripheral blood lymphocyte subtypes and its clinical significance. Methods: The pathologic and clinical data of 117 LELC patients who were admitted to the Tumor Hospital of the University of Chinese Academy of Sciences from 2006 to 2018 were collected. The histological classification was based on previously reported morphological classification method. The onco-immunological and morphologic characteristics of the tumors such as lymphoid follicle formation and interstitial fibrous hyperplasia, patient's peripheral blood lymphocyte subtypes and prognosis data were collected. The relationship between various factors and their impact on prognosis were analyzed. Results: There were 117 patients, including 61 females and 56 males. The male to female ratio was 0.9∶1.0. The age of onset was 24-89 years (median 52 years). Primary sites included head and neck (68 cases), lungs (26 cases), stomach (15 cases), and others (eight cases). Morphologically, 54 cases were type Ⅰ, 62 cases were type Ⅱ, and one case could not be classified. The onco-immunological and morphologic features of the LELC tumors showed a continuous spectrum. Interstitial TILs were noted from focally to diffuse, and the interstitial fibrous tissues were from hardly visible to obvious sclerotic. Formation of lymphoid follicles was seen in 42 patients; obvious fibrosis was seen in 31 cases. Data of peripheral blood lymphocyte subtyping by flow cytometry were available in 73 cases. These data included CD3+total T cells, CD3+CD4+helper T cells, CD3+CD8+cytotoxic T cells, CD3-CD56+natural killer (NK) cells, CD3-CD19+B cells, CD4+CD45RA-T helper induction subgroup, CD4+CD45RA+ T suppression induction subgroup, CD4+CD45RO+memory T cell subgroup, CD45RA+CD45RO+activated T cell subgroup, CD8+CD38+activated cytotoxic T cell, and CD25+lymphocytes and CD44+lymphocyte. The proportion of lymphocytes of each subtype was normal in most patients, but the proportion of CD44+lymphocytes in 61 cases (83.6%) was increased; the proportion of T cell suppression induced subgroups was decreased in 53 cases (72.6%). Correlation analysis found a significant correlation between clinical stage and NK cells (P=0.023); tumor histologic type and cytotoxic T cells were significantly positively correlated (P=0.012); while tumor cell morphologic differentiation was significantly related to total T cells (P=0.003) and NK cells (P=0.026); Formation of interstitial lymphoid follicles was positively correlated with memory T cell subsets (P=0.025); Tumor interstitial fibrosis was significantly positively correlated with T suppression-induced subpopulations (P=0.004), and was significantly negatively correlated with total T cells (P=0.023) and with the expression of CD44 adhesion molecules (P=0.003). Survival analysis found that lymphoid follicle formation was a favorable prognostic factor for LELC (P=0.001). Conclusions: The onco-immunological and morphologic features in LELC show a continuous spectrum; the tumor clinicopathological characteristics and onco-immunological morphology are closely related to peripheral blood T lymphocyte subtypes, and the formation of interstitial lymphoid follicles is a favorable prognostic factor for LELC.

[The prediction of disease progression by 18Fluorodeoxyglucose-positron emission computed tomography/CT in patients with dermatomyositis and interstitial lung disease].

To investigate the predictive value of [18F]fluorodeoxyglucose-positron emission computed tomography(PET)/CT for disease progression in patients with dermatomyositis (DM) and interstitial lung diseases (ILD). Sixty-seven DM patients who underwent [18F] FDG-PET/CT imaging were retrospectively analyzed from January 2012 to September 2017 at PLA General Hospital. Their clinical manifestations and imaging characteristics were recorded. Compared with those chronically progressed (C-ILD), patients with rapid progression (RP-ILD) had significantly higher erythrocyte sedimentation rate (ESR) and standardized uptake value (SUV) in lungs (P<0.05). In patients with RP-ILD, SUV in lungs was positively correlated with age, disease course, and ESR. Receiver operating characteristic curve analysis suggested that when lung SUV cut off value was 2.25, the sensitivity and specificity to predict disease progression was 77.8% and 72.8%, respectively. Old age, longer disease course, low creatine kinase level, higher ESR, and high SUV are prognostic factors for DM-associated ILD.

[The 478 th case: multiple myalgia with intramuscular nodules].

A 28-year-old man was admitted to the first medical center of Chinese People's Liberation Army General Hospital because of multiple myalgia and intramuscular nodules for more than 2 months. The patient complained of dysphagia, fever and weight loss 2 months ago. Magnetic resonance imaging and biopsy revealed nodular fasciitis. Inflammatory indicators including C-reactive protein, erythrocyte sedimentation rate, platelet count and fibrinogen were slightly elevated. Urine occult blood was positive. Abdominal ultrasound revealed left hydronephrosis. Because nodular fasciitis could not explain the whole situation, a needle biopsy of intramuscular nodules was performed. Pathological examination revealed intramuscular metastatic adenocarcinoma with poor differentiation. Gastric endoscope and positron emission tomography-computed tomography confirmed the diagnosis of advanced gastric adenocarcinoma with extensive metastases of esophagus, lymph nodes, muscles, ureter and bone. Although chemotherapy was given, the patient died of disease progression six months later.

[Family history of rheumatic diseases in patients with rheumatoid arthritis: a large scale cross-sectional study].

OBJECTIVE: To determine the associations between the family history of rheumatic diseases and clinical features in patients with rheumatoid arthritis (RA). METHODS: In total, eight hundred and ninety patients with RA were enrolled. The demographic and clinical data were collected, including gender, age, height, body weight, age of disease onset, history of smoking and drinking, family history of rheumatic diseases, clinical and laboratory features, pain and global visual analogue scale (VAS), and multi-dimensional health assessment questionnaire (MDHAQ). Finally, 803 patients were completed the dataset and were included in the study. RESULTS: In this cohort, the male/female ratio was 1:3.5, and the age of onset was (45.09±14.50) years. A total of 123 (15.32%) patients were accompanied with family history of rheumatic diseases, including RA, spondyloarthritis, Sjögren's syndrome, systemic lupus erythematosus and systemic sclerosis. The percentages of first degree, second degree and both first and second degree relatives were 91 (73.98%), 22 (17.89%), and 10 (8.13%) respectively. The most common disease was RA (70.73%), followed by other rheumatic diseases (21.95%), and RA combined with other rheumatic diseases (7.32%). The clinical and laboratory characteristics were compared between the patients with and without family history. The onset-age of the subjects was significantly different between those with and without family history of rheumatic diseases (39.97 ±13.68 vs. 46.01±14.46; P<0.01), which meant that the onset-age in patients with family history was 6.04 years earlier than that in patients without family history. The patients with family history had higher positive rate of rheumatoid factor (RF) compared with those without family history (78.48% vs. 66.67%, P<0.05). By adjusting with gender, body mass index (BMI), smoking and alcohol drinking, anti-cyclic citrullinated peptide (CCP) antibody and RF level, the age at disease onset in the patients with family history was 4.54 years earlier than that in the patients without family history (ß=-4.54; 95%CI:-8.70, -0.38; P<0.05). Further hierarchical regression analysis showed that, the age at onset of the RA patients with family history was 10.02 years earlier than that without family history among the smoking patients (ß= -10.02; 95%CI:-17.60, -2.43; P=0.01), while the age at onset of the RA patients with family history was 3.27 years earlier than that without family history among the never smoking patients (ß=-3.27; 95%CI:-8.37, 1.82; P=0.21). CONCLUSION: The family history of rheumatic diseases is a risk factor for early onset of RA, and may interact with smoking.

[Effect of GPX4 on proliferation and metastasis of renal clear cell carcinoma and its relationship with expression of IGF-1R and COX-2].

Objective: To investigate the effect of human glutathione peroxidase 4 (GPX4) on the proliferation and metastasis of renal clear cell carcinoma and its relationship with the expression of IGF-1R and COX-2. Methods: Culture of human normal tubular cell line HK-2 and human renal clear cell carcinoma Caki-1, A498, Caki-2, 786-o in vitro. Detection of GPX4 mRNA and protein expression in different cell lines by quantitative real-time PCR (RT-PCR) and Western blot assay. Overexpression of GPX4 cell lines, including blank carrier (Vector) and overexpress GPX4 (oeGPX4) group, and interference with GPX4 renal clear cell carcinoma cell lines, including random sequence (shControl), interference GPX4#1 (shGPX4#1) and interference GPX4#2 (shGPX4#2) group by lentiviral transfection. RT-PCR technology and Western blot were used to detect the expression of GPX4, IGF-1R and COX-2 mRNA and protein. CCK-8 assay was used to detect the relative proliferation of cells at 0, 24, 48, 72 and 96 h in each group. Transwell invasion and migration assay to detect the invasion and migration ability of cells of each group. Results: GPX4 is highly expressed in renal clear cell carcinoma cell lines compared to human normal tubular cell lines; The expression of GPX4, IGF-1R and COX-2 mRNA was significantly increased in oeGPX4 cells compared with Vector cells, the expression of GPX4,IGF-1R and COX-2 mRNA was significantly decreased in shGPX4#1 and shGPX4#2 compared with shControl cells; oeGPX4 cells significantly increased proliferative capacity compared to Vector cells at 72 and 96 h, the proliferation of shGPX4#1 and shGPX4#2 cells was significantly lower than that of shControl cells at 72 and 96 h; The number of invading and migrating cells of oeGPX4 cells was significantly higher than that of Vector cells, the number of invasive and migrating cells in shGPX4#1 and shGPX4#2 cells was significantly lower than that in shControl cells. Conclusion: GPX4 is highly expressed in renal clear cell carcinoma cells, which is positively correlated with the expression of IGF-1R and COX-2, and can promote cell proliferation and metastasis in vitro.

Mechanistic overview of immune checkpoints to support the rational design of their combinations in cancer immunotherapy.

Checkpoint receptor blockers, known to act by blocking the pathways that inhibit immune cell activation and stimulate immune responses against tumor cells, have been immensely successful in the treatment of cancer. Among several checkpoint receptors of immune cells, cytotoxic T-lymphocyte-associated protein-4 (CTLA-4), programmed cell death protein-1 (PD-1), T-cell immunoglobulin and ITIM domain (TIGIT), T-cell immunoglobulin-3 (TIM-3) and lymphocyte activation gene 3 (LAG-3) are the most commonly targeted checkpoints for cancer immunotherapy. Six drugs including one CTLA-4 blocker (ipilimumab), two PD-1 blockers (nivolumab and pembrolizumab) and three PD-L1 blockers (atezolizumab, avelumab and durvalumab) are approved for the treatment of different types of cancers including both solid tumors such as melanoma, lung cancer, head and neck cancer, bladder cancer and Merkel cell cancer as well as hematological tumors such as classic Hodgkin's lymphoma. The main problem with checkpoint blockers is that only a fraction of patients respond to the therapy. Insufficient immune activation is considered as one of the main reason for low response rates and combination of checkpoint blockers has been proposed to increase the response rates. The combination of checkpoint blockers was successful in melanoma but had significant adverse events. A combination that is selected based on the mechanistic differences between checkpoints and the differences in expression of checkpoints and their ligands in the tumor microenvironment could have a synergistic effect in a given cancer subtype and also have a manageable safety profile. This review aims to help in design of optimal checkpoint blocker combinations by discussing the mechanistic details and outlining the subtle differences between major checkpoints targeted for cancer immunotherapy.

[Septic arthritis in systemic lupus erythematosus: a retrospective analysis of 6 cases].

Base on the clinical characteristics of septic arthritis in a group of systemic lupus erythematosus patients, this study has found out that high systemic lupus erythematosus disease activity index, leucopenia, high cumulative dose of glucocorticoid, methylprednisolone intravenous pulse therapy and joint cavity puncture were closely correlated with septic arthritis. Once septic arthritis is suspected, culture specimens should be collected and appropriate antibiotics are suggested immediately. Also, surgical drainage is a very useful approach.

[The application of (18)F fluorodeoxyglucose-positron emission tomography/computed tomography in the diagnosis and treatment of dermatomyositis].

OBJECTIVE: To investigate the value of (18)F fluorodeoxyglucose-positron emission tomography/computed tomography ((18)F FDG-PET-CT) in the diagnosis and the evaluation of disease activity and remission of dermatomyositis(DM). METHODS: DM patients who were admitted to the Department of Rheumatology, the People's Liberation Army General Hospital (PLAGH) and underwent (18)F FDG-PET-CT examination were retrospectively reviewed from January 2012 to May 2015.Gender and age matched healthy controls (HC) were also enrolled.The standardized uptake value (SUV) of proximal limb girdle muscles in both groups were recorded and compared, so as between patients with DM or subclinical DM.The correlation between myodynamia of proximal limb girdle muscle, creatine kinase(CK), CK-MB, serum ferritin and SUV of each muscle group were analyzed. RESULTS: There were 58 patients with DM and 29 controls consecutively recruited in the study.The SUV of upper arms (1.814±0.830) g/ml, shoulders (2.134±0.797) g/ml and hips (1.883±0.683) g/ml in patients with classic DM were significantly higher than those with subclinical DM [(0.938±0.218) g/ml, (1.152±0.315) g/ml, (0.945±0.249) g/ml; P<0.05]. SUV of muscles in newly diagnosed patients was (1.051±0.031) g/ml, which was higher than that in subclinical patients.But the difference was not statistically significant (P>0.05). The average SUV of evaluated muscles in DM group (2.033±0.858) g/ml was significantly higher than that in controls (1.076±0.167) g/ml (P<0.001). Receiver operating characteristic(ROC) analysis revealed the area under the curve(AUC) of abnormal SUV detected by (18)F FDG-PET-CT for diagnosing DM was 0.953.The myodynamia of upper arms and SUV was negatively correlated (rs=-0.440, P=0.031). However, the level of serum creatine kinase and SUV was positively correlated (rs=0.500, P=0.013). The average SUV of patients whose time to remission was less than 3 months (1.746±0.466) g/ml was obviously less than that of patients with 3 to 6 months to obtain remission (2.815±0.848) g/ml (P=0.015). CONCLUSIONS: The SUV of proximal limb girdle muscles detected by (18)F FDG-PET-CT has a positive diagnostic value for DM.Moreover, the SUV in upper arms is correlated with the muscle strength and the level of creatine kinase, which reflect disease activity. (18)F FDG-PET-CT might be an alternative method to evaluate the response of treatment.

Isorhamnetin inhibits Prevotella intermedia lipopolysaccharide-induced production of interleukin-6 in murine macrophages via anti-inflammatory heme oxygenase-1 induction and inhibition of nuclear factor-κB and signal transducer and activator of transcription 1 activation.

BACKGROUND AND OBJECTIVE: Interleukin-6 (IL-6) is a key proinflammatory cytokine that has been considered to be important in the pathogenesis of periodontal disease. Therefore, host-modulatory agents directed at inhibiting IL-6 appear to be beneficial in terms of attenuating periodontal disease progression and potentially improving disease susceptibility. In the current study, we investigated the effect of the flavonoid isorhamnetin on the production of IL-6 in murine macrophages stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen implicated in inflammatory periodontal disease, and its mechanisms of action. MATERIAL AND METHODS: Lipopolysaccharide from P. intermedia ATCC 25611 was isolated using the standard hot phenol-water method. Culture supernatants were collected and assayed for IL-6. We used real-time PCR to quantify IL-6 and heme oxygenase-1 (HO-1) mRNA expression. The expression of HO-1 protein and the levels of signaling proteins were monitored using immunoblot analyses. The DNA-binding activity of nuclear factor-κB (NF-κB) was analyzed using ELISA-based assay kits. RESULTS: Isorhamnetin significantly down-regulated P. intermedia LPS-induced production of IL-6 as well as its mRNA expression in RAW264.7 cells. Isorhamnetin up-regulated the expression of HO-1 at both gene transcription and translation levels in cells stimulated with P. intermedia LPS. In addition, inhibition of HO-1 activity by tin protoporphyrin IX blocked the inhibitory effect of isorhamnetin on IL-6 production. Isorhamnetin failed to prevent LPS from activating either c-Jun N-terminal kinase or p38 pathways. Isorhamnetin did not inhibit NF-κB transcriptional activity at the level of inhibitory κB-α degradation. Isorhamnetin suppressed NF-κB signaling through inhibition of nuclear translocation and DNA binding activity of NF-κB p50 subunit and attenuated signal transducer and activator of transcription 1 signaling. CONCLUSION: Although further research is required to clarify the detailed mechanism of action, we propose that isorhamnetin may contribute to blockade of the host-destructive processes mediated by IL-6 and could be a highly efficient modulator of the host response in the treatment of inflammatory periodontal disease. Further research in animal models of periodontitis is required to better evaluate, the potential of isorhamnetin as a novel agent for treating periodontal disease.

Effects of Y27632 on keratinocyte procurement and wound healing.

A number of Rho-kinase inhibitors have been developed for various clinical applications. We examined the effects of the Rho-kinase inhibitor Y27632 on keratinocyte proliferation and migration, and found that it promoted primary human keratinocyte proliferation and migration in both monolayer and skin explant cultures. In addition, topical application of Y27632 enhanced cutaneous wound closure in the majority of wounds in mice. The growth and migration effects of Y27632 appeared to be specific to keratinocytes compared with dermal fibroblasts, and required intact Jun kinase function. Y27632 seems to be a promising agent for keratinocyte procurement and wounding healing.

Longitudinal Exposure-Response Modeling of Multiple Indicators of Alzheimer's Disease Progression.

BACKGROUND: Progression in Alzheimer's disease manifests as changes in multiple biomarker, cognitive, and functional endpoints. Disease progression modeling can be used to integrate these multiple measures into a synthesized metric of where a patient lies within the disease spectrum, allowing for a more dynamic measure over the range of the disease. OBJECTIVES: This study aimed to combine modeling techniques from psychometric research (e.g., item response theory) and pharmacometrics (e.g., hierarchical models) to describe the multivariate longitudinal disease progression for patients with mild-to-moderate Alzheimer's disease. Additionally, we aimed to extend the subsequent model to make it suitable for clinical trial simulation, with the inclusion of covariates, to explain variability in latent progression (i.e., disease progression) and to aid in the assessment of enrichment strategies. DESIGN: Multiple longitudinal endpoints in the Alzheimer's Disease Neuroimaging Initiative database were modeled. This model was validated internally using visual predictive checks, and externally by comparing data from the placebo arms of two Phase 2 crenezumab studies, ABBY (NCT01343966) and BLAZE (NCT01397578). SETTING: The Alzheimer's Disease Neuroimaging Initiative began in 2004: the initial 5-year study (ADNI-1) was extended by 2 years in 2009 by a Grand Opportunities grant (ADNI-GO), and in 2011 and 2016 by further competitive renewals of the ADNI-1 grant (ADNI-2 and ADNI-3, respectively). This work studies natural progression data from patients with confirmed Alzheimer's disease. The Phase 2 ABBY and BLAZE trials evaluated the safety and efficacy of crenezumab in patients with mild-to-moderate Alzheimer's disease. PARTICIPANTS: From the Alzheimer's Disease Neuroimaging Initiative database, 305 subjects who had a baseline diagnosis of mild-to-moderate Alzheimer's disease were included in modeling. From the ABBY and BLAZE studies, 158 patients were included from the studies' placebo arms. MEASUREMENTS: Longitudinal cognitive and functional assessments modeled included the Clinical Dementia Rating (both as Sum of Boxes and individual item scores), the Mini-Mental State Examination, the Alzheimer's Disease Assessment Scale - Cognitive Subscale, the Functional Activities Questionnaire, the Montreal Cognitive Assessment, and the Rey Auditory Verbal Learning Test. Also included were the imaging variable fluorodeoxyglucose-positron emission tomography and the following magnetic resonance imaging volumetrics: entorhinal, fusiform, hippocampal, intra-cranial, mid-temporal, ventricular, and whole brain. RESULTS: Applying item response theory approaches in this longitudinal setting showed clinical assessments informing a common disease scale in the following order (from early disease to late disease): Rey Auditory Verbal Learning Test, Functional Activities Questionnaire, Montreal Cognitive Assessment, Alzheimer's Disease Assessment Scale - Cognitive Subscale 12, Clinical Dementia Rating - Sum of Boxes, and Mini-Mental State Examination. The Clinical Dementia Rating communication and home-and-hobbies items were most informative at earlier disease stages, while memory, orientation, and personal care informed the disease status at later stages. A clinical trial simulation model was developed and accurately described within-sample longitudinal distribution of endpoints. Simplifying the model to use only baseline age, MMSE, and APOEε4 status as predictors, out-of-sample mean progression of ADAS-Cog and CDR Sum of Boxes in the ABBY and BLAZE placebo arms was accurately described; however, the variability in these endpoints was underpredicted and suggests possibility for further model refinement when extrapolating from the ADNI sample to trial data. Clinical trial simulations were performed to exemplify use of the model to investigate hypothetical disease modification effects on the multivariate, longitudinal progression on the Alzheimer's Disease Assessment Scale - Cognitive Subscale and the Clinical Dementia Rating - Sum of Boxes. CONCLUSIONS: The latent variable structure of item response theory can be extended to capture a variety of scales that are common assessments and indicators of disease status in mild-to-moderate Alzheimer's disease. These models are not intended to support causal inferences, but they do successfully characterize the observed correlation between endpoints over time and result in concise numerical indices of disease status that reflect the totality of evidence from considering the endpoints jointly. As such, the models have utility for a variety of tasks in clinical trial design, including simulation of hypothetical drug effects, interpolation of missing data, and assessment of in-sample information.

Anti-inflammatory effects and the underlying mechanisms of action of daidzein in murine macrophages stimulated with Prevotella intermedia lipopolysaccharide.

BACKGROUND AND OBJECTIVE: Host modulatory agents directed at inhibiting specific proinflammatory mediators could be beneficial in terms of attenuating periodontal disease progression and potentially enhancing therapeutic responses. The aim of this study was to investigate whether daidzein could modulate the production inflammatory mediators in macrophages stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen implicated in periodontal disease, and to delineate underlying mechanisms of action. MATERIAL AND METHODS: LPS was extracted from P. intermedia ATCC 25611 cells by the standard hot phenol-water method. The amounts of nitric oxide (NO) and interleukin-6 (IL-6) secreted into the culture medium were assayed. A real-time PCR was performed to quantify inducible nitric oxide synthase (iNOS) and IL-6 mRNA expression. We used immunoblot analysis to characterize iNOS protein expression, phosphrylation of c-Jun N-terminal kinase (JNK) and p38, degradation of inhibitory κB-α (IκB-α), nuclear translocation of nuclear factor-κB (NF-κB) subunits and phosphorylation of signal transducer and activator of transcription 1 (STAT1). The DNA-binding activity of NF-κB was assessed by using ELISA-based kits. RESULTS: Daidzein significantly inhibited the production of NO and IL-6, as well as their mRNA expression, in P. intermedia LPS-treated RAW264.7 cells. The JNK and p38 pathways were not involved in the regulation of LPS-induced NO and IL-6 release by daidzein. Daidzein inhibited the degradation of IκB-α induced by P. intermedia LPS. In addition, daidzein suppressed NF-κB transcriptional activity via regulation of the nuclear translocation and DNA-binding activity of NF-κB p50 subunit and blocked STAT1 phosphorylation. CONCLUSION: Although additional studies are required to dissect the molecular mechanism of action, our results suggest that daidzein could be a promising agent for treating inflammatory periodontal disease. Further research in animal models of periodontitis is necessary to better evaluate the potential of daidzein as a novel therapeutic agent to treat periodontal disease.

[Study on the effect of AIDS preventive cascade services provided by social organizations for men who have sex with men in China].

Objective: To analyze the effect and main gaps of each stage in the AIDS prevention cascade for men who have sex with men (MSM) provided in intervention projects supported by the China AIDS Fund for non-governmental organizations (CAFNGO) and provide suggestions to improve the quality of cascade services and project management. Methods: Data were collected through the CAFNGO management information system and field interviews to analyze the differences in the number of MSM receiving HIV testing and confirming tests, the newly reported patients, and the number of antiviral treatment (ART) referrals of newly established reported patients among different social organization service areas. A service chain chart was also drawn. Results: Between 2016 and 2020, 1 508 MSM intervention projects were funded by CAFNGO, including 1 183 234 MSM being mobilized to receive HIV testing. However, only 68.8% (1 183 234/1 719 139) of the testing capacity of social organizations was covered by these projects. As a result, 55 783 HIV-positive MSM were detected in preliminary screening, and only 86.6% (48 327/55 783) received confirming tests. The proportion of newly reported infections was 3.8% (45 347/1 183 234). The ratio of antiviral treatment (ART) referrals for newly reported patients between 2017 and 2020 was 89.8% (32 719/36 444). 75.8%(1 143/1 508) of total MSM intervention projects were implemented by community-based organizations (Non-registered civil affairs departments). In comparison, organizations registered in civil affairs departments took up 24.2% (365/1 508) of the total MSM intervention projects. No significant difference was noticed in the proportion of newly reported infected (3.8% and 3.8%) and the ratio of ART referrals (89.7% and 89.9%) between community-based organizations and registered organizations' projects. But these two proportions are significantly different between these two types of organizations in some areas in China. Conclusions: The AIDS prevention cascade established in CAFNGO has effectively promoted the early detection and treatment of infected MSM. However, CAFNGO needs more financial support to extend testing coverage for MSM. Meanwhile, confirmation testing for positives in preliminary screening and ART referrals needs to be improved for newly reported patients. In addition, various capacity building needs to be provided for different social organizations.