Objective: To investigate the pattern of lymph node metastasis (LNM) in patients with locally advanced (T₃, T₄) laryngeal squamous cell carcinoma (LALSC) and provide reference for the delineation of clinical target volume. Methods: Clinical data of 272 patients with LALSC treated in our hospital from 2000 to 2017 were retrospectively analyzed. All patients underwent bilateral neck dissection (at least level Ⅱ-Ⅳ). The LNM ratio of each node level was calculated. The risk factors of LNM were identified by univariate and multivariate logistic regression analyses. Results: LNM was found in 156 of 272 patients (57.1%). According to the location of primary lesions, all patients were divided into group A (n=72; unilateral without midline involvement), group B (n=86; unilateral with midline involvement) and group C (n=114; giant or central). In group A, the LNM ratio at ipsilateral level Ⅱ, Ⅲ and Ⅳ was 36.3%, 26.4% and 6.9%, whereas 13.9%, 8.3% and 1.4% at the contralateral level, respectively. In group B, the LNM ratio at ipsilateral level Ⅱ, Ⅲ and IV was 1.9%, 29.1% and 11.6%, whereas 18.6%, 14.0% and 1.2% at the contralateral level, respectively. In group C, the LNM ratio at the left neck level Ⅱ, Ⅲ and Ⅳ was 24.6%, 23.7% and 2.6%, whereas 21.9%, 26.3% and 6.1% at the right neck, respectively. Bilateral LNM ratio did not significantly differ between group A and group B/C (15.3%, 25.0%, P=0.093). Ipsilateral level Ⅲ metastasis (OR=2.929, 95%CI 1.041-8.245, P=0.042) and clinical N stage (OR=0.082, 95%CI 0.018-0.373, P=0.001) were associated with contralateral LNM. Ipsilateral level Ⅱ(P=0.043) or Ⅲ(P=0.009) metastasis were risk factors of the ipsilateral level Ⅳ metastasis. Conclusions Neck levels Ⅱ and Ⅲ are the high-risk LNM regions, whereaslevels Ⅳ and V are the low-risk areas. Ipsilateral level Ⅱ or Ⅲ metastases are the risk factors of ipsilateral level Ⅳ and contralateral cervical LNM. Contralateral neck LNM rarely occurs in cN₀ stage patients.

Objective To investigate the clinical,biochemical and genetic features of hereditary hypomagnesaemia with secondary hypocalcaemia.Methods Two boys came from different Chinese families.They were hospitalized at the Peking University First Hospital between 2014 and 2016 at the age of 9 years and 1 year and 2 months because of epilepsy and psychomotor retardation.Clinical investigation,laboratory examination,and medical imaging were performed for the etiological study.Whole-genome sequencing was used for the genetic analysis of the patients.Mutations of TRPM6 gene were confirmed by means of Sanger sequencing.Results Patient 1 presented with recurrent seizures and psychomotor retardation from the age of 3 months.Vision loss and psychomotor regression were noticed from the age of 9 years,accompanied with hypertension.Serum magnesium and total calcium were significandy decreased to 0.13-0.15 mmol/L and 1.43-2.00 mmol/L,respectively in patient 1.Serum potassium was reduced to 1.85-3.25 mmol/L.Blood parathyroid hormone was also decreased.On the TRPM6 gene of patient 1,2 novel non-sense mutations,c.2771G ＞ A (p.Trp924Ter) and c.115C ＞ T (p.Gln39Ter) were identified.Patient 2 presented with seizures and psychomotor retardation at the age of 2 weeks.Both of his serum magnesium (0.17-0.35 mmol/L) and serum total calcium (1.32-1.34 mmol/L) were significantly decreased.Blood parathyroid hormone was decreased.Two novel mutations (c.1239G ＞ A,p.W413X and c.146G ＞ A,p.C49Y) were found in the TRPM6 gene of patient 2.Severe hypomagnesaemia,hypocalcaemia and TRPM6 gene mutations confirmed the diagnosis of hereditary hypomagnesaemia with secondary hypocalcaemia in the 2 patients.After the large-dose supplement of magnesium sulfate,progressive clinical improvements were observed in the 2 patients.However,because of the severe brain damage,patient 1 still had psychomotor retardation.Patient 2 completely recovered.Conclusions Hereditary hypomagnesaemia with secondary hypocalcaemia is a severe inherited metabolic disease.Early diagnosis and large-dose magnesium supplement are the key to the good prognosis of the patients.In this study,2 Chinese children with the clinical onset of epilepsy and psychomotor retardation are reported.The diagnosis is made by way of blood biochemical assay and gene analysis.Four novel mutations on their TRPM6 gene are identified.

Objective To investigate the relationship of wall motion abnormality and myocardium viability with chronic total occlusion (CTO) lesions by cardiac magnetic resonance (CMR) imaging. Methods We identified 128 patients with CTO lesion of at least one coronary artery confirmed by coronary arteriography at Beijing Anzhen Hospital between December 2014 to November 2017. All of the patients received CMR examination after admission. We analyzed the CMR images according to the AHA/American College of Cardiology 17-segment model, and recorded the left ventricular ejection fraction (LVEF), end-systolic volume (ESV) and end-distolic volume (EDV) calculated by CMR. Results In the myocardium regions corresponding to the 149 CTO lesions, only 11.5％ presented transmural myocardial infarction. A 58.6％ of the myocardial segments showed no delayed enhancement. Patients with delayed enhancement degree over 75％ had the lowest LVEF and the largest EDV and ESV. Conclusion CMR showed that patients with CTO lesions had different degrees of myocardial infarction. Patients with transmural myocardial infarction had significant cardiac function decline and ventricular remodeling. Only a fraction of patients had transmural infarction, suggesting that a majority of the CTO patients would benefit from treatment.

The mammalian central nervous system (CNS) is considered an immune privileged system as it is separated from the periphery by the blood brain barrier (BBB). Yet, immune functions have been postulated to heavily influence the functional state of the CNS, especially after injury or during neurodegeneration. There is controversy regarding whether adaptive immune responses are beneficial or detrimental to CNS injury repair. In this study, we utilized immunocompromised SCID mice and subjected them to spinal cord injury (SCI). We analyzed motor function, electrophysiology, histochemistry, and performed unbiased RNA-sequencing. SCID mice displayed improved CNS functional recovery compared to WT mice after SCI. Weighted gene-coexpression network analysis (WGCNA) of spinal cord transcriptomes revealed that SCID mice had reduced expression of immune function-related genes and heightened expression of neural transmission-related genes after SCI, which was confirmed by immunohistochemical analysis and was consistent with better functional recovery. Transcriptomic analyses also indicated heightened expression of neurotransmission-related genes before injury in SCID mice, suggesting that a steady state of immune-deficiency potentially led to CNS hyper-connectivity. Consequently, SCID mice without injury demonstrated worse performance in Morris water maze test. Taken together, not only reduced inflammation after injury but also dampened steady-state immune function without injury heightened the neurotransmission program, resulting in better or worse behavioral outcomes respectively. This study revealed the intricate relationship between immune and nervous systems, raising the possibility for therapeutic manipulation of neural function via immune modulation.

PURPOSE: The chemical structure of tubulosine has been known since the mid-1960s. However, little is known about its biological and pharmacological functions. The aim of this study was to investigate the novel functions of tubulosine in cancer treatment, specifically in breast cancer. METHODS: An Unpaired (Upd)-induced Drosophila cell line and interleukin (IL)-6-stimulated human breast cancer cell lines were used to investigate the biological and pharmacological activities of tubulosine in vitro. To investigate the activities of tubulosine, we performed molecular and cellular experiments such as Western blot and reverse transcription polymerase chain reaction analyses, immunoprecipitation and terminal deoxynucleotidyl transferase dUTP nick end labeling assays, and immunofluorescence staining using breast cancer cell lines. RESULTS: Tubulosine exhibited anticancer activity in IL-6-stimulated human breast cancer cells. Moreover, tubulosine reduced the tyrosine phosphorylation level and transcriptional activity of signal transducer and activator of transcription (STAT) protein at 92E in Upd-induced Drosophila cells. Additionally, tubulosine suppressed IL-6-induced Janus kinase 2 (JAK2)/STAT3 signaling, resulting in decreased viability and induction of apoptotic cell death in breast cancer cells. Interestingly, inhibition of IL-6-induced JAK2/STAT3 signaling by tubulosine was associated with the blocking of IL-6 receptor (IL-6R) and glycoprotein 130 (gp130) binding. CONCLUSION: Tubulosine exhibits anticancer activity through functional inhibition of IL-6-induced JAK2/STAT3 signaling by targeting IL-6Rα/gp130 binding in breast cancer cells. These findings suggest that tubulosine may hold promise for the treatment of inflammation-associated cancers, including breast cancer.
Blotting, Western ; Breast Neoplasms ; Cell Death ; Cell Line ; DNA Nucleotidylexotransferase ; Drosophila ; Fluorescent Antibody Technique ; Glycoproteins ; Humans ; Immunoprecipitation ; In Vitro Techniques ; Interleukin-6 ; Interleukins ; Janus Kinase 2 ; Phosphorylation ; Phosphotransferases ; Polymerase Chain Reaction ; Receptors, Interleukin-6 ; Reverse Transcription ; STAT3 Transcription Factor ; Transducers ; Tyrosine

PURPOSE: Rituximab with cyclophosphamide, doxorubicin, vincristine, and prednisone administered every 3 weeks (R-CHOP-21) is the standard care for diffuse large B-cell lymphoma (DLBCL). It is unknown whether the dose-dense R-CHOP (R-CHOP-14) could improve the outcome of the disease in Asian population. MATERIALS AND METHODS: Newly diagnosed DLBCL patients were centrally, randomly assigned (1:1) to receive R-CHOP-14 or R-CHOP-21. R-CHOP-14 was administered every 2 weeks, and R-CHOP-21 was administered every 3 weeks. Primary end point was disease-free survival (DFS). Secondary end points included overall survival (OS), progression-free survival (PFS), response rate and toxicities. RESULTS: Seven hundred and two patients were randomly assigned to receive R-CHOP-14 (n=349) or R-CHOP-21 (n=353). With a median follow-up of 45.6 months, the two groups did not differ significantly in 3-year DFS (79.6% for R-CHOP-14 vs. 83.2% for R-CHOP-21, p=0.311), 3-year OS (77.5% for R-CHOP-14 vs. 77.6% for R-CHOP-21, p=0.903), or 3-year PFS (63.2% for R-CHOP-14 vs. 66.1% for R-CHOP-21, p=0.447). Patients with an International Prognostic Index (IPI) score ≥ 2 had a poorer prognosis compared to those with an IPI score < 2. Grade 3/4 hematologic and non-hematologic toxicities were manageable and similar between R-CHOP-14 and R-CHOP-21. CONCLUSION: R-CHOP-14 did not improve the outcome of DLBCL compared to R-CHOP-21 in Asian population. With manageable and similar toxicities, both of the two regimens were suitable for Asian DLBCL patients. For high-risk patients with IPI ≥ 2, new combination regimens based on R-CHOP deserve further investigation to improve efficacy.
Asian Continental Ancestry Group ; B-Lymphocytes ; Cyclophosphamide ; Disease-Free Survival ; Doxorubicin ; Follow-Up Studies ; Humans ; Lymphoma, B-Cell ; Prednisone ; Prognosis ; Rituximab ; Vincristine

To establish a novel colloidal gold immunochromatography assay (GICA) for rapid, sensitive and accurate detection of Haemophilus influenzae infection by using the outer membrane protein P6 as detection target. First, the linear antigen epitope located in the extracellular domain of the P6 protein (GenBank accession number: AGH02799) was predicted by bioinformatics analysis. The region (62-75 aa of the protein) with strong antigen specificity was chosen and synthesized. Two rabbits were then immunized by the polypeptides (14 aa) for production of polyclonal antibodies. Then, the recombinant P6 proteins were also obtained to produce polyclonal antibodies. Finally, based on the two antibodies, a novel colloidal GICA for detection of Haemophilus influenzae infection was established and the specificity, sensitivity, repeatability and stability of this method were evaluated. At the same time, the method was tested in clinical simulation, and the plate culture method was used to verify its accuracy. The test strip for Haemophilus influenzae infection was successfully prepared. The detection limit of the test strip was as low as 1×105 CFU/mL and the whole process can be completed within 15 minutes. The strip specifically recognized Haemophilus influenzae and did not react with nine of other common respiratory pathogens such as Streptococcus pneumoniae, Moraxella catarrhalis, Mycoplasma pneumonia, and Legionella pneumophila. And the strips could be stored at 25 °C for at least 6 months without losing sensitivity or specificity. The coincidence rate between the results of 200 clinical samples and the plate culture method was 90.5%. Haemophilus influenzae protein P6, which possessed a high degree of surface antigen accessibility and antigencity, could be used as a marker for Haemophilus influenzae detection. The immunochromatographic colloidal gold test strip which bears the features of rapidity, convenience and sensitivity provides a unique tool for the on-site surveillance and diagnosis of Haemophilus influenzae infection in clinical test.
Animals ; Chromatography, Affinity ; instrumentation ; Diagnostic Tests, Routine ; standards ; Gold Colloid ; chemistry ; Haemophilus Infections ; diagnosis ; Haemophilus influenzae ; Humans ; Limit of Detection ; Rabbits ; Sensitivity and Specificity

To study the effects of saikosaponin b2( SS-b2) on inflammatory factors and energy metabolism against lipopolysaccharide/galactosamine( LPS/Gal N) induced acute liver injury in mice. Mice were randomly divided into normal group( equal amount of normal saline),model group( 100 g·kg~(-1) LPS and 400 mg·kg~(-1) Gal N),low,medium,high dose group of SS-b2( SS-b25,10,20 mg·kg~(-1)·d-1) and positive control group( dexamethasone,10 mg·kg~(-1)). All of the groups except for the normal group were treated with LPS/Gal N though intraperitoneally injection to establish the acute liver injury model. The organ indexes were calculated. The levels of serum transaminases( ALT and AST) and the activities of ATPase( Na+-K+-ATPase,Ca2+-Mg2+-ATPase) in liver were detected. The activity of tumor necrosis factor-α( TNF-α),interleukin-1β( IL-1β) and interleukin-6( IL-6) were determined by the enzyme-linked immunosorbent assay( ELISA). The contents of lactate dehydrogenase( LDH) in liver were determined by micro-enzyme method. HE staining was used to observe the histopathological changes of the liver. Histochemical method was used to investigate the protein expression of liver lactate dehydrogenase-A( LDH-A). The protein expressions of Sirt-6 and NF-κB in the liver were detected by Western blot. According to the results,compared with the model group,there were significant changes in organ indexes in the high-dose group of SS-b2( P<0. 05). The level of ALT,AST,TNF-α,IL-1β,IL-6 and the activities of LDH in serum of mice with liver injury were significantly reduced in the medium and high dose groups of SS-b2( P<0. 01). With the increase of the concentration of SS-b2,the range of hepatic lesions and the damage in mice decreased. The activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in liver of mice were significantly enhanced in each dose group( P<0. 01). The expression of NF-κB in liver tissues was significantly down-regulated in the medium and high dose group( P<0. 01). Meanwhile,the expression of Sirt-6 protein in the liver of mice with acute liver injury was significantly increased in each dose group( P<0. 01).In summary,SS-b2 has a significant protective effect on LPS/Gal N-induced acute liver injury in mice,which may be related to the down-regulation of NF-κB protein expression and up-regulation of Sirt-6 protein expression to improve inflammatory injury and energy metabolism.
Animals ; Chemical and Drug Induced Liver Injury ; drug therapy ; Cytokines ; metabolism ; Energy Metabolism ; Galactosamine ; Inflammation ; drug therapy ; Lipopolysaccharides ; Liver ; drug effects ; Mice ; NF-kappa B ; metabolism ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Random Allocation ; Saponins ; pharmacology ; Sirtuins ; metabolism

OBJECTIVE: To evaluate the effect of intranasal immunization with CTA1-DD as mucosal adjuvant combined with H3N2 split vaccine. METHODS: Mice were immunized intranasally with PBS (negative control), or H3N2 split vaccine (3 μg/mouse) alone, or CTA1-DD (5 μg/mouse) alone, or H3N2 split vaccine (3 μg/mouse) plus CTA1-DD (5 μg/mouse). Positive control mice were immunized intramuscularly with H3N2 split vaccine (3 μg/mouse) and alum adjuvant. All the mice were immunized twice, two weeks apart. Then sera and mucosal lavages were collected. The specific HI titers, IgM, IgG, IgA, and IgG subtypes were examined by ELISA. IFN-γ and IL-4 were test by ELISpot. In addition, two weeks after the last immunization, surivival after H3N2 virus lethal challenge was measured. RESULTS: H3N2 split vaccine formulated with CTA1-DD could elicit higher IgM, IgG and hemagglutination inhibition titers in sera. Furthermore, using CTA1-DD as adjuvant significantly improved mucosal secretory IgA titers in bronchoalveolar lavages and vaginal lavages. Meanwhile this mucosal adjuvant could enhance Th-1-type responses and induce protective hemagglutination inhibition titers. Notably, the addition of CTA1-DD to split vaccine provided 100% protection against lethal infection by the H3N2 virus. CONCLUSION CTA1-DD could promote mucosal, humoral and cell-mediated immune responses, which supports the further development of CTA1-DD as a mucosal adjuvant for mucosal vaccines.
Adjuvants, Immunologic ; Administration, Intranasal ; Animals ; Cholera Toxin ; Female ; Immunity, Humoral ; Influenza A Virus, H3N2 Subtype ; immunology ; Influenza Vaccines ; Mice, Inbred BALB C ; Nasal Mucosa ; immunology ; Random Allocation ; Recombinant Fusion Proteins

To study the correlation between ultra high performance liquid chromatography( UPLC) fingerprint of Smilax china and its anti-pelvic inflammatory effect,and to explore the pharmacodynamic material basis of S. china against pelvic inflammatory disease.UPLC fingerprints of 10 batches of S. china from different habitats were established,and the values of SOD,MDA,TNF-α,and IL-6 in rats with pelvic inflammation were measured. The weight of each single pharmacodynamics index to the total efficacy was determined by analytic hierarchy process,and the contribution of each peak in fingerprints to the each single pharmacodynamics index and total efficacy was analyzed by the grey relational analysis. Then the structures of chemical constituents at the identified peaks were confirmed by comparing with the reference substance. The 27 common characteristic peaks of UPLC fingerprints were all related to the anti-pelvic inflammation effect of S. china,of which 13 peaks were identified as peak 2( 3,5-dihydroxy-2-methylbenzoic acid-3-O-glucoside),peak 3( chlorogenic acid),peak 5( 2,7,4-trihydroxydihydroflavone-5-O-glucoside),peak 6( 7,4-dihydroxydihydroflavonol-5-O-glucoside),peak 7( taxifolin-7-O-glucoside),peak 9( taxifolin),peak 10( polydatin),peak 11( oxyresveratrol),peak 12( astilbin),peak15( resveratrol),peak 16( quercitrin),peak 18( engeletin) and peak 24( kaempferol). The correlation degree of 21 peaks and the total efficacy was greater than 0. 8,and the top 10 ranked by correlation degree were as follows: peak 1,3,7,19,18,17,4,11,16,and 21. The results showed that the anti-pelvic inflammation effect of S. china was achieved by the combined action of pharmacodynamic substances. In order to control the quality of S. china and its prepared slices more effectively,the index components of content detection should be selected reasonably.
Animals ; China ; Chromatography, High Pressure Liquid ; Female ; Pelvic Inflammatory Disease ; drug therapy ; Phytochemicals ; pharmacology ; Plant Extracts ; pharmacology ; Rats ; Smilax ; chemistry