[Regulatory Mechanism of Mangiferin Combined with Bortezomib on Malignant Biological Behavior of Burkitt Lymphoma and Its Effect on Expression of CXC Chemokine Receptors].

OBJECTIVE: To analyze the effects of mangiferin combined with bortezomib on the proliferation, invasion, apoptosis and autophagy of human Burkitt lymphoma Raji cells, as well as the expression of CXC chemokine receptors (CXCRs) family, and explore the molecular mechanism between them to provide scientific basis for basic research and clinical work of Burkitt lymphoma. METHODS: Raji cells were intervened with different concentrations of mangiferin and bortezomib alone or in combination, then cell proliferation was detected by CCK-8 assay, cell invasion ability was detected by Transwell chamber method, cell apoptosis was detected by Annexin V/PI double-staining flow cytometry, apoptosis, autophagy and Akt/mTOR pathway protein expression were detected by Western blot, and the expression changes of CXCR family was detected by real-time quantitative PCR (RT-qPCR). RESULTS: Different concentrations of mangiferin intervened Raji cells for different time could inhibit cell viability in a concentration- and time-dependent manner (r =-0.682, r =-0.836). When Raji cells were intervened by combination of mangiferin and bortezomib, compared with single drug group, the proliferation and invasion abilities were significantly decreased, while the apoptosis level was significantly increased (P <0.01). Mangiferin combined with bortezomib could significantly up-regulate the expression of pro-apoptotic protein Bax and down-regulate the expression of anti-apoptotic protein Bcl-2 after intervention in Raji cells. Caspase-3 was also hydrolyzed and activated, and then induced the apoptosis of Raji cells. Mangiferin combined with bortezomib could up-regulate the expression of LC3Ⅱ protein in Raji cells, and the ratio of LC3Ⅱ/LC3Ⅰ in cells was significantly up-regulated compared with single drug or control group (P <0.01). Mangiferin combined with bortezomib could significantly inhibit the phosphorylation levels of Akt and mTOR, inhibit the proliferation and invasion of Raji cells by inhibiting Akt/mTOR pathway, and induce cell autophagy and apoptosis. Mangiferin and bortezomib could down-regulate the expressions of CXCR4 and CXCR7 mRNA after single-agent intervention in Raji cells, and the down-regulations of CXCR4 and CXCR7 mRNA expression were more significant when the two drugs were combined (P <0.01). Mangiferin alone or combined with bortezomib had no significant effect on CXCR5 mRNA expression in Raji cells (P >0.05), while the combination of the two drugs could down-regulate the expression of CXCR3 (P <0.05). CONCLUSION: Mangiferin combined with bortezomib can synergistically inhibit the proliferation and invasion of Raji cells, and induce autophagy and apoptosis. The mechanism may be related to the inhibition of Akt/mTOR signaling pathway, down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bax, and the inhibition of the expression of CXCR family.

Photocatalytic C-C Bond Cleavage and Fluorosulfonylation of Strained Cycloalkanols for Carbonyl-Containing Aliphatic Sulfonyl Fluorides.

In this report, we present a photocatalytic ring-opening fluorosulfonylation of strained cycloalkanols with sulfur dioxide and NFSI under mild conditions for the synthesis of carbonyl-containing aliphatic sulfonyl fluorides. The synthetic potential of the carbonyl-containing aliphatic sulfonyl fluoride products has been examined by diverse transformations, including SuFEx reactions and Baeyer-Villiger oxidation reactions. Mechanistic studies demonstrate that the reaction operates through a radical C-C bond cleavage/SO2 insertion/fluorination cascade process.

Aminofluorosulfonylation of ß,γ-Unsaturated Hydrazones with Sulfur Dioxide and N-Fluorobenzenesulfonimide: Accessing Pyrazoline-Functionalized Aliphatic Sulfonyl Fluorides.

An efficient aminofluorosulfonylation strategy was developed for the synthesis of various pyrazoline-functionalized aliphatic sulfonyl fluorides using ß,γ-unsaturated hydrazones with sulfur dioxide and NFSI as the starting materials under mild conditions. The sulfonyl fluoride products could be successfully transformed into the corresponding sulfonate esters and amides via the sulfur(VI) fluoride exchange (SuFEx) click reactions. Preliminary mechanistic investigations demonstrate that the reaction operates through a radical cyclization/SO2 insertion/fluorination cascade process.

[The Value of T Cell subsets and Cytokine Levels Changes in the Clinical Diagnosis, Treatment and Prognosis Evaluation of Multiple Myeloma].

OBJECTIVE: To explore the correlation between the changes of T lymphocyte subsets and cytokines in patients with MM and immune function status, biochemical indicators, and their relationships with clinical stage and prognosis, which is expected to provide a scientific basis for the prognosis analysis and condition monitoring of MM patients. METHODS: The clinical data of 89 MM patients in two hospitals were collected, and 36 healthy people without tumor or infectious diseases were selected as the control group. Flow cytometry and enzyme-linked immunosorbent assay (ELISA) were used to detect the changes of core members of peripheral blood T lymphocyte subsets and cytokine levels, respectively. At the same time, automatic biochemical analyzer and automatic blood cell analyzer were used to detect serum ß2-microglobulin (ß2-MG), lactate dehydrogenase (LDH), albumin (ALB), creatinine (CRE) and hemoglobin (HGB) levels, and the relationship between T lymphocyte subsets and the above indexes and their clinical significance were analyzed. RESULTS: The proportions of NK cells and CD8+T lymphocytes in the peripheral blood of MM patients were significantly higher than that of the control group (P<0.01), the proportion of CD4+T and the ratio of CD4+/CD8+ were lower than those of the control group (P<0.05); however, there was no significant difference in the numbers CD3+T cells compared with the control group (P>0.05). The proportion of CD4+T and ratios of CD4+/CD8+ in MM patients were lower than those of normal controls, and were negatively correlated with MM staging (r=-0.964, r=-0.653), that is, the later the MM staging, the more obvious their levels were reduced, while CD8+T and NK cells were positively correlated with MM staging (r=0.891, r=0.728), that is, the later the MM staging, the more significant their levels increased. The levels of Treg cells (CD4+CD25highCD127low/-T cells/CD4+T cells) of MM patients in the disease stage Ⅰ, Ⅱ and Ⅲ were (5.87±0.92)%, (7.97±1.32)%, (11.52±4.71)% respectively, the difference was statistically significant compared with control group (P<0.05), and the level of Treg cells in MM patients with stage III was significantly higher than that in controls and patients with other disease stages (P<0.01). The proportion of Treg cells (CD4+CD25highCD127low/-T cells/CD4+T cells) in MM patients was positively correlated with the concentration of ß2-MG and LDH (r=0.793, r=0.536), but had no significant correlation with HGB, ALB and CRE. The serum levels of IL-6, IL-10 and TNF-α in MM patients were significantly higher than those in the control group (P<0.05), which were closely related to MM staging(r=0.839, r=0.917, r=0.746), that is, the later the MM staging, the higher the levels; The serum IFN-γ level was negatively correlated with the stage of MM (r=-0.689), and its level gradually decreased with the increase of the disease stage and degree (P<0.01). There was no significant correlation between the levels of IL-2 and IL-4 and the disease stage, but they were all up-regulated compared with the control group (P<0.05). CONCLUSION: The abnormal regulation of the core members of T lymphocyte subsets and the levels of various cytokines are closely related to the disease progression and poor prognosis of MM patients, which is an effective indicator for the disease monitoring of MM patients.

NIS-Promoted Selective Amino-Diazidation and Amino-Iodoazidation of O-Homoallyl Benzimidates: Synthesis of Vicinal Diazido 1,3-Oxazines and Vicinal Iodoazido 1,3-Oxazines.

Amines, especially those with multi-nitrogen moieties, are widespread in natural products and biologically active compounds. Thus, the development of direct and efficient methods to introduce multiple nitrogen-containing fragments into compounds in one step is highly desirable yet challenging. Herein, we report an NIS-promoted selective amino-diazidation and amino-iodoazidation of O-homoallyl benzimidates with NaN3. By using this protocol, a variety of vicinal diazido-substituted 1,3-oxazines and vicinal iodoazido-substituted 1,3-oxazines were directly synthesized in a controllable manner. Preliminary mechanistic investigations revealed that the reaction operates through a NIS-promoted four-step cascade process. The developed method has the merits of metal-free, excellent functional group compatibility, simple operation, and mild conditions.

[Diagnostic accuracy of MALDI-TOF mass spectrum in identification of oral Candidiasis isolates].

PURPOSE: To investigate the diagnostic accuracy of matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) in the identification of clinical oral Candida isolates. METHODS: In the first experiment, 170 suspected oral candidiasis patients were included. Unstimulated saliva was collected and cultured on solid plate (Sabouraud's agar) for 48 hours. Clinical isolates were analyzed by both PCR, which was used as golden standard, and MALDI-TOF MS. The diagnostic accuracy of MALDI-TOF MS for both C.albicans and non-C.albicans were investigated. In the second experiment, we tried to identify Candida isolates directly from positive liquid culture in a new cohort of 42 patients. MALDI-TOF MS identification of candida isolates were performed on positive liquid medium cultures directly. The data were analyzed using SPSS 18.0 software package. RESULTS: Totally, 230 clinical Candida isolates were isolated from 212 oral candidiasis patients enrolled. Out of the 230 analyzed clinical isolates, C. albicans (65.65%, 151/230) was the most frequently isolated species, followed by C. glabrata (11.74%, 27/230), and other candida species. The sensitivity and specificity of MALDI-TOF in identification of C. albicans were 93.33% and 92.73% respectively, and those for non-C. albicans were 83.64% and 89.2% respectively. Oral rinse followed by culturing in Sabouraud's liquid medium for 24 hours yield the diagnostic accuracy of 78.42% in MALDI-TOF MS identification. CONCLUSIONS: C. albicans is the most frequent Candida spp. in oral candidiasis. MALDI-TOF MS is an effective and rapid method for identification of Candida spp. The on-plate based method generate much higher diagnostic accuracy than liquid culture based one.

Efficacy of nystatin for the treatment of oral candidiasis: a systematic review and meta-analysis.

OBJECTIVE: To systematically review and assess the efficacy, different treatment protocols (formulation, dosage, and duration), and safety of nystatin for treating oral candidiasis. METHODS: Four electronic databases were searched for trials published in English till July 1, 2015. Randomized controlled trials comparing nystatin with other antifungal therapies or a placebo were included. Clinical and/or mycological cure was the outcome evaluation. A meta-analysis or descriptive study on the efficacy, treatment protocols, and safety of nystatin was conducted. RESULTS: The meta-analysis showed that nystatin pastille was significantly superior to placebo in treating denture stomatitis. Nystatin suspension was not superior to fluconazole in treating oral candidiasis in infants, children, or HIV/AIDS patients. The descriptive investigations showed that administration of nystatin suspension and pastilles in combination for 2 weeks might achieve a higher clinical and mycological cure rate, and using the nystatin pastilles alone might have a higher mycological cure rate, when compared with using nystatin suspensions alone. Nystatin pastilles at a dose of 400,000 IU resulted in a significantly higher mycological cure rate than that administrated at a dose of 200,000 IU. Furthermore, treatment with nystatin pastilles for 4 weeks seemed to have better clinical efficacy than treatment for 2 weeks. Descriptive safety assessment showed that poor taste and gastrointestinal adverse reaction are the most common adverse effects of nystatin. CONCLUSION: Nystatin pastille was significantly superior to placebo in treating denture stomatitis, while nystatin suspension was not superior to fluconazole in treating oral candidiasis in infants, children, or HIV/AIDS patients. Indirect evidence from a descriptive study demonstrated that administration of nystatin pastille alone or pastille and suspension in combination is more effective than that of suspension alone; prolonged treatment duration for up to 4 weeks can increase the efficacy of nystatin. More well designed and high quality randomized control studies are needed to confirm these findings.

[Validation of the American College of Rheumatology classification criteria for primary Sjogren's syndrome in Chinese patients].

OBJECTIVE: To validate the American College of Rheumatology Classification (ACR) Criteria (2012) for the diagnosis of primary Sjogren's syndrome in Chinese patients. METHODS: All patients involved in this study came from the Department of Oral Medicine Peking University Stomatology Hospital. They were devided into two groups of pSS and non-pSS according to the diagnoses made by two experts. Both groups of the patients had completed medical records kept in the hospital. RESULTS: A total of 239 pSS patients (160 with labial salivary gland biopsy, and 79 without biopsy) and 52 age-matched non-pSS patients [(55.17±14.295),and (55.90±13.38) years old, P>0.05] (9 with biopsy, and 43 without biopsy) were involved in this study. The sensitivity and specificity of ACR criteria in diagnosing pSS were 90.37 % and 88.46 % respectively. The positive and negative likelihood ratios were 7.83 and 0.109, respectively. The sensitivities of ACR criteria in diagnosing pSS patients with and without labial biopsy were 88.75% and 93.67 %, respectively, with specificities of 88.89% and 88.37%, respectively. The most sensitive item adopted in ACR criteria was the ocular staining score with a sensitivity of 85.77%, and the most specific item was the labial salivary gland biopsy, with a specificity of 88.89%. CONCLUSION: The sensitivity and specificity of ACR criteria in diagnosing Chinese pSS patients were relatively high, and may serve as the diagnosis criteria in research and clinical practice. However, the ACR criteria need to be validated and further revised in the future .

[Detection of oral fungal flora in patients with primary Sjogren's syndrome by real-time PCR].

OBJECTIVE: To compare the quantity of the main important Candida species in the oral cavity between the patients with primary Sjogren's syndrome (pSS) and no dry mouth healthy controls,and to explore the discriminative species of fungi between the two groups. METHODS: In this study, 25 pSS patients from Department of Oral Medicine of Peking University School and Hospital of Stomatology were enrolled, with 25 residents from a community in Beijing as the control group. All the participants were examined for the oral mucosa status, and 5 Candida DNA loads were compared between the two groups by use of the FQ-PCR technique. RESULTS: Candida albicans was the dominant Candida in both of the groups. In addition, we found that the species of Candida tropicalis and Parapsilosis were more detected in the pSS group by use of the FQ-PCR technique. Quantitative of Candida tropicalis and Candida parapsilosis in pSS group (10.6 ± 1.07, 9.47 ± 4.86) was significantly higher than that in healthy controls group (8.30 ± 3.82, 5.24 ± 6.05), the difference was statistically significant (P <0.05). Quantitative of fluconazole-sensitive strains in pSS group (12.21 ± 0.82) compared with which in healthy controls group (11.53 ± 0.81) was significantly increased (P <0.01). CONCLUSION: Candida albicans was the dominant Candida in both of the groups. Candida tropicalis and Candida parapsilosis were more detected in the pSS group.

[Association of HAb18G with clinicopathologic features and prognosis in non-small cell carcinoma of lung].

OBJECTIVE: To study the association between HAb18G expression, tumor parameters, metastatic potential and prognosis in non-small cell lung carcinoma (NSCLC). METHODS: Immunohistochemical study for HAb18G protein using SP methods was carried out in 144 cases of NSCLC. Nineteen cases of benign lung lesions and 41 cases of normal lung tissue were used as controls. The intensity (positive unit/PU) of HAb18G expression was assessed quantitatively by image analysis software. The results were correlated with tumor parameters, metastatic potential and follow-up data. RESULTS: The intensity of HAb18G protein expression was significantly higher in NSCLC than that in controls (P = 0.000). In squamous cell carcinomas and adenocarcinomas, the expression of HAb18G protein in well-differentiated tumors was lower than that in moderately to poorly differentiated tumors (P = 0.001). Tumors of TNM stage IV had stronger expression than tumors of lower stages (P = 0.000). HAb18G PU was greater in tumors with lymph node metastasis than those without nodal metastasis (P = 0.045). The PU value of tumors with maximal diameter greater than 5 cm was higher than that of the smaller tumors (P = 0.000). It was also higher in male than in female patients (P = 0.046). There was no association between HAb18G protein expression and age of patients, history of smoking, tumor types and gross morphology (P > 0.05). The five-year survival rate in cases with low HAb18G protein expression was higher than that in cases with high expression (P = 0.006). Univariate analysis indicated that patients with high HAb18G protein expression carried a poor prognosis (P = 0.007). Multivariate analysis showed that expression of HAb18G protein was an independent prognostic factor in patients with NSCLC (P = 0.032, relative risk 3.962). CONCLUSIONS: HAb18G protein expression is associated with tumor progression and prognosis. It may represent a useful biomarker for prognostic evaluation.

[Preliminary study of the relationship between tumor like Sjögren's syndrome and malignant lymphoma].

OBJECTIVE: To investigate the clinical and laboratory characteristics of tumor like Sjögren's syndrome (TLSS) patients and non-tumor like Sjögren's syndrome (NTLSS) and the incidence of lymphoma in patients of Sjögren's syndrome (SS). METHODS: A retrospective analysis was carried out in 199 primary SS (including TLSS) patients who were recruited in Peking University School and Hospital of Stomatology from 1998 to 2010. Clinical and laboratory information were collected. The patients were divided into two groups: TLSS (n = 25) and NTLSS (n = 174). Clinical and laboratory characteristics were compared between these two groups by a statistical analysis. RESULTS: Of the 25 TLSS patients, 23 had enlargements of parotid glands and 2 had enlargements of submandibular glands. There were significant differences of salivary scintigraphy appearance (P = 0.018), hypergammaglobulinemia (P = 0.014), rheumatoid factor positive rate (P = 0.001), formation of the ectopic germinal centers (P = 0.014), double positive rate of anti-SSA antibody and anti-SSB antibody (P < 0.001) between the TLSS and NTLSS patients. Among the 25 TLSS patients, 3 developed lymphomas, accounting for 1.5% (3/199) of the total 199 patients and 12% (3/25) of the TLSS patients. Lymphoma subtypes included one diffused large B-cell lymphoma and two mucosa-associated lymphoid tissue lymphoma. There was no lymphoma detected in NTLSS patients. CONCLUSIONS: There are clinical and laboratory differences between TLSS and NTLSS patients, with a more tendency to develop lymphomas in TLSS patients.

[Clinical assessment of oral diagnostic items in 2002 classification criteria for primary Sjogren's syndrome].

OBJECTIVE: To evaluate the diagnostic value of each item in 2002 international classification criteria for primary Sjogren's syndrome (pSS) in clinically diagnosed cases. METHODS: All patients were from the Department of Oral Medicine of Peking University School of Stomatology from 2005 to 2010. Their clinical manifestations and lab testing results met the standard criteria and the diagnosis was established according to international classification criteria (2002). We retrospectively collected all of the clinical and information and did the data analysis. RESULTS: A total of 148 pSS patients were included in the study. When the oral evaluations were performed, 98.0% of the patients complained dry mouth, 96.6% of them had decreased non-stimulated salivary flow rate, and 60.1% had positive parotid sialography results, which was consistent with diagnosis. It was found out in the lab testing that 79.7% of the patients had positive anti-SSA antibody, and 75.0% of them had elevated globulin level. 20.3% of the patients were finally diagnosed by performing the lower lip biopsy. CONCLUSION: When applying 2002 international classification criteria for the diagnosis of pSS patients, oral symptoms and unstimulated salivary flow rate are feasible and sensitive, which play an important and indicative role in the diagnosis of pSS.

[Development of an immunochromatographic strip test for rapid detection of Vibrio vulnificus].

OBJECTIVE: To establish a rapid strip test for detection of Vibrio vulnificus. METHOD: Anti-Vibrio vulnificus polyclonal antibodies were obtained from the rabbits immunized with Vibrio vulnificus (ATCC27562). Colloidal gold was prepared through reducing HAuCl(4)·4H(2)O by sodium citrate and conjugated with the polyclonal antibodies as the detecting reagent. The polyclonal antibodies and sheep anti-rabbit immunoglobulins were separately coated onto the same nitrocellulose membrane for sample detection and quality-control, respectively. The nitrocellulose membrane, gold conjugate pad, sample pad, filter paper and absorbent pad were assembled to prepare the strips. The detection specificity and sensitivity of this strip were evaluated. RESULTS: The strip test for detecting Vibrio vulnificus yielded results in 20 to 30 min. The detection sensitivity of the test was 2×10(6) CFU/ml. The strip showed no cross-reaction with other bacterial strains. The strips remained stable after preservation at 4 degrees celsius; for 4 months. CONCLUSION: With a high specificity and sensitivity, this strip test is applicable in the detection of Vibrio vulnificus.

[Clinical and pathological analysis of oral manifestations of 40 patients with secondary syphilis].

OBJECTIVE: To analyze the clinical and pathological features of 40 patients with secondary syphilis. METHODS: A total of 40 cases of secondary syphilis confirmed by serology were collected during 1994-2004 and were first diagnosed on presentation with oral lesions. RESULTS: The white patch in oral mucosa was found in 32 cases with painless or slight pain in most cases. The most common site of the lesion was the tongue. The histological examination on eight cases was initially misdiagnosed as oral candidosis or lichen planus, but confirmed as syphilis after serology revealed nonspecific inflammation with intraepithelial microabscess and dense perivascular infiltration of lymphocytes and plasma cells in connective tissue. The symptoms showed dramatic improvement in 16 cases after benzathine penicillin treatment. CONCLUSIONS: The oral manifestations of syphilis have specific clinical and pathological feature and attention should be paid to the suspicious oral lesions when patients are first presented in a dental office.

[Histopathological features of oral pemphigus vegetans].

OBJECTIVE: To investigate the clinicopathological features of oral pemphigus vegetans. METHODS: Seven cases of pemphigus vegetans involving oral mucosa were included in this study. The paraffin sections were analyzed by routine light microscope. RESULTS: There were 5 females and 2 males in this group, with an average age of 52.7 years. The course of diseases was from 3 months to 10 years. Oral manifestations included verrucous or papilliform hyperplasia of oral mucosa, with extensive edema and exudation. Similar skin lesions were found in 5 patients. Under the microscope, pemphigus vegetans showed epithelial thickening with prolonged rete pegs, intraepithelial clefts in suprabasal cells. Eosinophilic microabscesses was often seen within the epithelium. Other microscopic changes were intraepithelial edema and eosinophils infiltration, chronic inflammatory cells infiltration in the lamina propria. CONCLUSION: The microscopic features pemphigus vegetans described in this study are of valuable for pathological diagnosis of oral pemphigus vegetans.