Suppressor of cytokine signaling 1 (SOCS1) silencing and Hep-2 sensitizing dendritic cell vaccine in laryngocarcinoma immunotherapy.

OBJECTIVE: Many studies have recently suggested that dendritic cell (DC) vaccine contributes to the immunotherapy of various types of human tumors. It has been proved that the tumor antigen sensitizing and the gene silencing are effective methods for the preparation of the DC vaccines. The aim of this study is to investigate the specific anti-laryngocarcinoma immune response for the suppression of cytokine signaling1 (SOCS1) silencing and Hep-2 sensitizing DC. MATERIALS AND METHODS: The dendritic cells derived from peripheral blood mononuclear cells were induced by cytokines GM-CSF, IL-4, and TNF-α in vitro, and the morphological characteristics of dendritic cells were observed under a microscope, indicating that they successfully differentiated into dendritic cells. The RNA interference vector was used to transfect dendritic cells. The expression of SOCS1 was detected by Western blot and the effective target sequence for inhibiting the expression of SOCS1 was screened. The expressions of CD83, CD86, and HLA-DR on dendritic cells were detected by flow cytometry. The content of IFN-γ in the supernatant was analyzed by enzyme-linked immunosorbent assay (ELISA). Methyl thiazolyl tetrazolium (MTT) was used to evaluate the ability of dendritic cells to stimulate T cell proliferation and induce the killing activity of cytotoxic T cells. RESULTS: The result of PCR and Western blot analysis shows that the expression of SOCS1 significantly decreased under the influence of the 5th interference sequence. The flow cytometric analysis results show that SOCS1 silencing and Hep-2 sensitizing dendritic cells had high expressions of CD83 (85.61±0.96)%, CD86 (96.86±1.20)%, and HLA-DR (98.02±0.94)%. The DC vaccine could increase the production of IFN-γ according to the ELISA assay results. The MTT assay results show that the DC vaccine could also stimulate the proliferation of the T cells and effectively and eventually enhance the specific killing effect of CTL. CONCLUSIONS: SOCS1 silencing and Hep-2 sensitizing DC vaccine could induce an effective and specific anti-laryngocarcinoma immune response.

[Efficacy and mechanism of loading dose clopidogrel in patients with transient ischemic attack and minor stroke].

Objective: To assess outcome, safety and possible mechanism of loading dose clopidogrel in patients with transient ischemic attack (TIA) and minor stroke. Methods: We reviewed patients with confirmed TIA and minor stroke admitted between July 2016 and December 2017 into the First Affiliated Hospital of Soochow University. Loss-of-function allele carriers of CYP2C19 were included and randomly divided into loading dose group (first dose of 300 mg clopidogrel) and standard dose group (first dose of 75 mg clopidogrel), 100 mg aspirin was gave at the same time, followed by aspirin 100 mg/d plus clopidogrel 75 mg/d maintaining for 20 days. Platelet aggregation (maximum aggregation ratio, MAR) induced by Adenosine diphosphate (ADP) was examined before and 3 days after administration. The National Institutes of Health Stroke Scale (NIHSS) score method was employed to assess the NIHSS scores before and after treatment in each group of patients; the modified Rankin Scale (mRS) was used to assess the 3-month functional outcome. Results: There was no significant difference in baseline data between the two groups (P>0.05).The proportion of early neurological function improvement in the two groups was 75.0% and 54.8%, and the difference was statistically significant (χ(2)=4.498, P=0.034). The 3-month prognosis was 79.5% and 61.3%, and the difference was statistically significant (χ(2)=4.000, P=0.045). Adverse events: 1 case in the loading dose group, 1 case in the standard dose group, the difference was not statistically significant (2.3% vs 1.6%, χ(2)=0.061, P=0.806). After 3 days of antiplatelet therapy, the MAR of the loading dose group decreased (11%±8%), and the MAR of the standard dose group decreased (9%±4%), the difference was statistically significant (P=0.013).In the loading dose group, there were 32 (72.7%)CYP2C19*2 carriers and 42 (95.5%)CYP2C19*2+*3 carriers; early neurological function improvement in 33 cases, accounting for 93.8% and 76.2%, respectively, and the difference was statistically significant (χ(2)=4.122, P=0.042). There were 35 patients with good prognosis in 3 months, accounting for 96.9% and 81.0%, respectively. The difference was statistically significant (χ(2)=4.310, P=0.038); MAR of CYP2C19*2 carrier was decreased (15%±5%), and MAR of CYP2C19*2+*3 carrier was decreased (12%±8%). The difference was statistically significant (P=0.039). Conclusions: Loading dose clopidogrel can improve the clinical prognosis of minor stroke/TIA without increasing the risk of bleeding. Loading dose clopidogrel may improve the prognosis of minor stroke/TIA by decreasing MAR of CYP2C19*2 carriers.

Immunoexpression of aquaporins 1, 2, 3 and 4 in kidney of yak (Bos grunniens) on the Qinghai-Tibetan Plateau.

Aquaporins (AQP) 1, 2, 3 and 4 belong to the aquaporin water channel family and play an important role in urine concentration by reabsorption of water from renal tubule fluid. Renal AQPs have not been reported in the yak (Bos grunniens), which resides in the Qinghai Tibetan Plateau. We investigated AQPs 1-4 expressions in the kidneys of Yak using immunohistochemical staining. AQP1 was expressed mainly in the basolateral and apical membranes of the proximal tubules and descending thin limb of the loop of Henle. AQP2 was detected in the apical plasma membranes of collecting ducts and distal convoluted tubules. AQP3 was located in the proximal tubule, distal tubule and collecting ducts. AQP4 was located in the collecting ducts, distal straight tubule, glomerular capillaries and peritubular capillaries. The expression pattern of AQPs 1-4 in kidney of yak was different from other species, which possibly is related to kidney function in a high altitude environment.

[Research progress of xerostomia assessment tools].

Xerostomia is an important problem affecting oral health and quality of life, and effective assessment is the basis of management of xerostomia. This review summarizes the subjective and objective assessment tools for xerostomia, so as to provide references for the evaluation and management of xerostomia.

B vitamin treatments modify the risk of myocardial infarction associated with a MTHFD1 polymorphism in patients with stable angina pectoris.

BACKGROUND: Methylenetetrahydrofolate dehydrogenase (MTHFD1) catalyzes three sequential reactions that metabolize derivatives of tetrahydrofolate (THF) in folate-dependent one-carbon metabolism. Impaired MTHFD1 flux has been linked to disturbed lipid metabolism and oxidative stress. However, limited information is available on its relation to the development of atherothrombotic cardiovascular disease. METHODS AND RESULTS: We explored the association between a MTHFD1 polymorphism (rs1076991 C > T) and acute myocardial infarction (AMI), and potential effect modifications by folic acid/B12 and/or vitamin B6 treatment in suspected stable angina pectoris patients (n = 2381) participating in the randomized Western Norway B Vitamin Intervention Trial (WENBIT). During the median follow-up of 4.9 years 204 participants (8.6%) suffered an AMI. After adjusting for established CVD risk factors, the MTHFD1 polymorphism was significantly associated with AMI (HR: 1.49; 95% CI, 1.23-1.81). A similar association was observed among patients allocated to treatment with vitamin B6 alone (HR: 1.53; 95% CI, 1.01-2.31), and an even stronger relationship was seen in patients treated with both vitamin B6 and folic acid/B12 (HR: 2.35; 95% CI, 1.55-3.57). However, no risk association between the MTHFD1 polymorphism and AMI was seen in patients treated with placebo (HR: 1.29; 95% CI, 0.86-1.93) or folic acid/B12 (1.17; 95% CI, 0.83-1.65). CONCLUSION: A common and functional MTHFD1 polymorphism is associated with increased risk of AMI, although the risk seems to be dependent on specific B vitamin treatment. Further studies are warranted to elucidate the possible mechanisms, also in order to explore potential effect modifications by nutritional factors.

Association between polymorphisms in the adiponectin gene (APM-1) and atherosclerotic cerebral infarction in a Hainan Chinese Han population.

We investigated the association between polymorphisms in the adiponectin gene (APM-1) and atherosclerotic cerebral infarction (ACI) in a Chinese Han population of Hainan Province. Polymerase chain reaction-restriction fragment length polymorphism and gene sequencing were used to analyze the distribution of APM-1 +45T/G and +276G/T genotypes and their alleles in 120 ACI patients and 120 healthy controls. No statistical correlation was found in the frequency and distribution of the genotype 45T/G between the ACI group and the control group. Genotypic frequencies of GG, GT, and TT at the APM-1 +276 locus were 70.0% (84/120), 25.0% (30/120), and 5.0% (6/120), respectively, in the ACI group, while these values were 52.5% (63/120), 37.5% (45/120), and 10.0% (12/120), respectively, in the control group. The frequency of the G allele was 82.5% (198/240) in the ACI group and 71.25% (171/240) in the control group. The T allele frequency was 17.5% (42/240) in the ACI group and 28.75% (69/240) in the control group. Polymorphisms at the APM-1 -276 locus in the case-controlled groups showed significant differences in the genotype distribution and al-lele frequency between the 2 groups (P = 0.041). The occurrence of ACI in the Hainan Chinese Han population may be associated with +276G/T polymorphisms but not with +45T/G polymorphisms in the APM-1 gene.

Prognostic value of clinical and immunological markers in acute phase of SFTS virus infection.

SFTS virus (SFTSV) is a novel bunyavirus that causes severe fever with thrombocytopenia syndrome (SFTS), an emerging infectious disease that occurred in China in recent years, with an average case fatality rate of 10-12%. Intervention in the early clinical stage is the most effective measure to reduce the mortality rate of disease. To elucidate the natural course of and immune mechanisms associated with the pathogenesis of SFTSV, 59 laboratory-confirmed SFTS patients in the acute phase, who were hospitalized between October 2010 and September 2011, were enrolled in this study, and the patients sera were dynamically collected and tested for SFTSV viral RNA load, 34 cytokines or chemokines and other related laboratory parameters. All clinical diagnostic factors in the acute phase of SFTS were evaluated and assessed. The study showed that the severity of the disease in 11 (18.6%) patients was associated with abdominal pain (p 0.007; OR = 21.95; 95% CI, 2.32-208.11) and gingival bleeding (p 0.001; OR=122.11; 95% CI, 6.41-2328). The IP-10, TNF-α, IL-6, IL-10, granzyme B and HSP70 levels were higher over the 7-8 days in severe cases, accompanied by altered AST, CK and LDH levels. HSP70 (p 0.012; OR=8.29; 95% CI, 1.58-43.40) was independently correlated with the severity of the early acute phase of SFTSV infection. The severity of SFTS can be predicted based on the presence of symptoms such as abdominal pain and gingival bleeding and on the level of HSP70 in the acute phase of the disease.

Telbivudine treatment is associated with high hepatitis B e antigen seroconversion and immune modulatory effects in chronic hepatitis B patients.

Chronic hepatitis B (CHB) is characterized by an impaired immune response to hepatitis B virus. Among the nucleos(t)ides used in CHB treatment, telbivudine is associated with the highest rates of hepatitis B e antigen (HBeAg) seroconversion rates, which are similar to those observed with pegylated interferon (PegIFN). Besides direct antiviral effect, modulation of the immune system may be an additional benefit for telbivudine-treated patients. Indeed, there is much clinical data indicating an IFN-like behaviour for telbivudine in contrast to other oral nucleos(t)ides, such as high HBeAg seroconversion, similar hepatitis B surface antigen (HBsAg) decline and biphasic viral kinetics. Clinical studies, animal models and in vitro studies suggest that both the innate and adaptive immune system responses contribute to high HBeAg seroconversion during telbivudine treatment through modulation of the function and/or expression of CD4+/CD8+ T cells, Th1/Th2, Treg, PD-1/PD-L1, Th17, IL-21 and TFH. The results described in this review suggest that the antiviral effect of telbivudine may be attributable not only to direct suppression of hepatitis B virus, but also to immunoregulatory effects. Hypothetically, telbivudine shares some common signal pathways with IFN.

Rapid downregulation of programmed death-1 and interferon-γ-inducible protein-10 expression is associated with favourable outcome during antiviral treatment of chronic hepatitis B.

The dynamics of programmed death-1 (PD-1) as well as cytokine/chemokine expression and its correlation with virological response in patients with chronic hepatitis B (CHB) is unclear. This study was conducted in 29 treatment-naïve patients undergoing telbivudine treatment for 52 weeks. The results showed that PD-1 expression on both CD4+ and CD8+ T cells was positively correlated with hepatitis B virus (HBV) DNA levels (r = 0.621, P < 0.0001; r = 0.548, P = 0.002, respectively), and in virological responders, this decrease was directly correlated with a decrease in HBV DNA levels (r = 0.664, P = 0.002; r = 0.572, P = 0.01, respectively). Furthermore, at the end of 52 weeks, in virological responders, the decreased rate in the frequency of PD-1+ CD8+ T cells was significantly higher than in non-virological responders (58.3% vs 25.7%, P = 0.001), and at weeks 24 and 52, in virological responders, PD-1 expression on CD4+ and CD8+ T cells was lower than in non-virological responders (P = 0.01 and P = 0.035; P < 0.0001 and P < 0.0001, respectively). In 34 cytokines/chemokines detected in serum, IP-10 expression was positively correlated with viral load, level of ALT and PD-1 expression on CD8+ and CD4+ T cells at baseline (r = 0.36, P = 0.055, r = 0.635, P < 0.0001, r = 0.414, P = 0.026, and r = 0.402, P = 0.030, respectively). Moreover, the decrease in IP-10 in serum directly correlated with a decrease in ALT levels (r = 0.751, P < 0.0001). At weeks 24 and 25, IP-10 expression was significantly lower than baseline in virological responders (both P = 0.005); however, this was not observed in nonresponders. Based on the above findings, PD-1 and IP-10 may be used as predictors for virological response, and blockade of their pathway may improve the outcome of patients with CHB.

The cranial cervical ganglion and its branches in the White yak (Bos grunniens).

The heads of 12 White yaks (four castrated, four male and four female, 3-8 years old) were dissected to study the shape, location and branches of the cranial cervical ganglion macroscopically. The ganglion was a greyish arciform structure, with a mean length of 17.3 mm, a width of 8.0 mm and a thickness of 3.9 mm, located on the rostrolateral surface of m. longus capitis. Approximately 5% of the ganglion was covered laterally by the tympanic bulla and the rest by the m. stylohyoideus. The branches of the cranial cervical ganglion included the internal and external carotid nerves, the sympathetic trunk and communicating branches to the glossopharyngeal nerve, the pharyngeal branch of the vagus nerve and the hypoglossal nerve. In one specimen, the left cranial cervical ganglion was fusiform and only covered by the m. stylohyoideus. Gender differences of the cranial cervical ganglion in the White yak were not observed.

Relationship between hypertension and hearing disorders in the elderly.

OBJECTIVE: To investigate the relationship between hypertension and hearing disorders of the elderly. DESIGN: Prospective case-control study. SETTING: Lin Yi People's Hospital, Shandong Province, the People's Republic of China, from October 1995 to June 1997. PARTICIPANTS: Fifty five hypertensive patients and fifty five normal elderly subjects (control group) between the ages of fifty five and eighty nine years old were enrolled in the study. All patients were male and were divided into three groups according to age as follows: Group A: 55-64; Group B: 65-74; Group C: 75-89. INTERVENTIONS AND MAIN OUTCOME MEASURES: SIMENS DA-64 audiometer and MEDSON 2250 auditory brainstem response audiometry (ABR) were utilised to determine the degree of hearing disorders. Serum triglycerides and cholesterol were also tested in our central laboratory. RESULTS: Auditory thresholds collected during audiometric tests increased gradually in proportion with age, especially in the hypertensive group (p < 0.05). The results of the ABR demonstrated that the latencies of wave V, I-VIPL and III-VIPL were also prolonged compared with normal elderly subjects. The patient's duration of illness and the complications of hypertension had an influence on hearing disorders in relation to the ageing process. CONCLUSION: Hypertension, with a high level of blood triglycerides and cholesterol, worsens the hearing of the elderly. The hearing disorder in elderly people is a result of long duration of illness and the complications of the hypertension.

[Demonstration on Z-D phenomenon in the occurrence of infectious diseases].

A systematic analysis, including retrospective test by means of correlation analysis and extrapolation test was carried out to demonstrate a new hypothesis named Z-D phenomenon which states that the skew of a seasonal case distribution curve of an infectious disease may appear to be the omen of its epidemic trend in the following year. A total number of 353 data sets belong to 17 notifiable infectious diseases in 29 provinces and the whole nation were chosen. Data from retrospective test showed that 91.2% (322/353) of the correlation coefficients of between the skew of the seasonal distribution curve of the previous year and the morbidity variation of its following year were negative. The extrapolation test showed that 68.1% (203/295) of the data sets accorded with the actually epidemic situation at the level of r < -0.5. The results proved the existence of Z-D phenomenon may serve as an indicator for the purpose of forecasting. Using this systematic analysis, we found that Z-D phenomenon was more expressive in viral hepatitis, pertussis, epidemic cerebrospinal meningitis and scarlet fever than in other diseases.

Enhanced anti-tumor effect of an IFN-gamma-EGF fusion protein.

The novel fusion proteins harboring human or mouse interferon combined with epidermal growth factor receptor binding domain were constructed using methods of genetic and protein engineering. The fusion proteins were assayed to retain complete antiviral activities. The EGF receptor binding moiety of the fusion proteins exhibited competitive binding against 125I-EGF for EGF receptors on A431 cells. The fusion proteins were shown to be more potent in inhibiting the growth of cultured target carcinoma cells than interferon-gamma alone. Experimental data derived from mouse B16 malignant melanoma models indicates that the weight of tumor in mice treated with IFN fusion proteins was significantly smaller than that of mice treated with interferon-gamma alone. The work here is unprecedented in the world and provides a reliable evidence to support the upcoming clinical employment of a class of interferons that specifically target tumor cell.

Localization of the gene responsible for the op (osteopetrotic) defect in rats on chromosome 10.

Osteopetrosis, a skeletal disorder of inadequate bone resorption with an abnormal increase in skeletal mass, results from a variety of independent single gene mutations that affect osteoclast differentiation and/or function. The osteopetrotic defect, op, is one of four spontaneous, nonallelic mutations in rats that result in osteopetrosis. In intercross progeny of (BN/SsN x LEW/SsN. +/op) F1 carriers, we mapped this locus by linkage analysis with microsatellite markers to rat chromosome 10. The linkage group contained, as well as op, 15 anonymous DNA loci and 9 DNA loci associated with genes (interleukin-3, myosin heavy chain [skeletal, embryonic], asialoglycoprotein receptor [hepatic lectin]-1, vesicle-associated membrane protein [synaptobrevin-2], sex hormone binding globulin, aldolase C, nitric oxide synthase [inducible], erythroblastic leukemia avian viral oncogene homolog-2, and proline-rich protein). The markers for these loci include nine not previously reported. The op locus mapped to the end of the chromosome 10 linkage group, within 1 cM of the anonymous DNA locus, D10Mit6. Based on its location, the op gene is likely to be distinct from seven described mutations in mice as well as three other mutations in rats. These results may permit a positional cloning strategy to be undertaken to identify the gene and mutation underlying the op defect.

Genetic maps of polymorphic DNA loci on rat chromosome 1.

Genetic linkage maps of loci defined by polymorphic DNA markers on rat chromosome 1 were constructed by genotyping F2 progeny of F344/N x LEW/N, BN/SsN x LEW/N, and DA/Bkl x F344/Hsd inbred rat strains. In total, 43 markers were mapped, of which 3 were restriction fragment length polymorphisms and the others were simple sequence length polymorphisms. Nineteen of these markers were associated with genes. Six markers for five genes, gamma-aminobutyric acid receptor beta3 (Gabrb3), syntaxin 2 (Stx2), adrenergic receptor beta1 (Adrb1), carcinoembryonic antigen gene family member 1 (Cgm1), and lipogenic protein S14 (Lpgp), and 20 anonymous loci were not previously reported. Thirteen gene loci (Myl2, Aldoa, Tnt, Igf2, Prkcg, Cgm4, Calm3, Cgm3, Psbp1, Sa, Hbb, Ins1, and Tcp1) were previously mapped. Comparative mapping analysis indicated that a large portion of rat chromosome 1 is homologous to mouse chromosome 7, although the homologs of two rat genes are located on mouse chromosomes 17 and 19. Homologs of the rat chromosome 1 genes that we mapped are located on human chromosomes 6, 10, 11, 12, 15, 16, and 19.

Linkage maps of rat chromosomes 15, 16, 17, 19, and X.

Linkage maps of rat chromosomes 15, 16, 17, 19, and X were constructed by multipoint genetic linkage analysis of 22 polymorphic markers in 40 F2 progeny of Fischer (F344/N) and Lewis (LEW/N) inbred rat strains. These markers are associated with eight genes (angiotensin receptor A, M3 muscarinic acetylcholine receptor, heme oxygenase, endothelin receptor A, haptoglobin, tyrosine aminotransferase, phosphoribosylpyrophosphate synthetase subunit II, and 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase) and 14 anonymous loci. Linkage analysis placed the markers into five linkage groups covering 11.7,7.9,11.6,42.5, and 5.1cM. These linkage groups were assigned to rat chromosomes 15, 16, 17, 19, and X, respectively, either by mouse x rat somatic cell hybrid analysis or based on previously identified locations of severalloci. In polymorphism analysis, these markers exhibited two to nine different alleles in 16 inbred rat strains.

[Levels of VIIIR:Ag, VIII:C and fibrinogen in plasma of patients with lung cancer].

Levels of VIIIR: Ag, VIII: C and fibrinogen (Fbg) in plasma have been measured in 80 patients with primary lung cancer. The results demonstrate that the plasma levels of VIIIR: Ag, VIII:C and Fbg in patients with lung cancer increase significantly in comparison with normal control group (P < 0.001). The levels increase along with the disease progressing. After treatment, all the three levels have decreased in treatment effective group. It is concluded that the measurements of plasma levels of VIIIR: Ag, VIII:C and Fbg are important in judging the condition of patients, to prognose and instruct the treatment for patients with lung cancer.