Objective: To observe the effect of needling Hegu (LI 4) on functions of the prefrontal cortex using the functional near infrared spectroscopy (fNIRS). Methods: A total of 12 healthy volunteers were randomly assigned to an acupuncture group (n=6) and a control group (n=6). The control group received no intervention. The acupuncture group received needling Hegu (LI 4) on the right side. The real-time concentration changes of oxygenated hemoglobin (HbO2), deoxygenated hemoglobin (d-Hb) and total hemoglobin (t-Hb) in the prefrontal cortex were recorded using the fNIRS, allowing for effect analysis of needling Hegu (LI 4) on the functions of the prefrontal cortex. Results: Compared with the control group, the HbO2 and t-Hb concentrations were increased in the acupuncture group and there were between-group statistical significances (bothP<0.05). There was no between-group statistical significance in d-Hb concentration (P>0.05). Conclusion: Needling Hegu (LI 4) can extensively activate the prefrontal cortex and therefore has significant effect on the functions of the brain's prefrontal cortex.

OBJECTIVETo establish an immunological method for detecting antibodies of Penicillium marneffei.

METHODSThe recombinant Mp1p protein of Penicillium marneffei was expressed in Pichia pastoris and labeled with HRP (Mp1p-HRP) with a modified sodium periodate method. A double-antigen sandwich enzyme-linked immunosorbant assay (ELISA) was established by determining the optimal coating concentration of Mp1p protein and the concentration of the detecting protein Mp1p-HRP. The sensitivity and specificity of the assay was evaluated by detecting Mp1p antibodies in 100 serum samples from healthy donors, 15 samples from culture-confirmed penicilliosis patients, and 21 samples from patients with culture-confirmed other fungal infections.

RESULTSA double-antigen sandwich ELISA was successfully established for detecting Mp1p-specific antibody. The specificity of the assay was 100% (121/121) for detecting Mp1p-specific antibody in the sera from healthy donors and patients with other fungal infection. The detection results of the 15 serum samples from patients with culture-confirmed penicilliosis showed positivity for Mp1p antibody in 2 samples and Mp1p antigen positivity in 12 samples; combining the detection results of Mp1p antigen and antibody obviously increased the diagnostic sensitivity to 93.3% (14/15).

CONCLUSIONThe double-antigen sandwich ELISA shows a high specificity in detecting Mp1p-specific antibody, and simultaneous detection of Mp1p antigen and antibody can increase the diagnostic sensitivity for penicilliosis.

Antibodies, Fungal ; blood ; immunology ; Antigens, Fungal ; blood ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Mycoses ; blood ; diagnosis ; microbiology ; Penicillium ; immunology ; Pichia ; immunology ; Sensitivity and Specificity

OBJECTIVE： To establish a method for content determination of N-（E）-p-coumaroyltyrosine in leaves of Abrus cantoniensis， and to optimize its purification technology. METHODS： HPLC method was adopted for the content determination of N-（E）-p-coumaroyltyrosine in A. cantoniensis. The determination was performed on Hypersil BDS C18 column with mobile phase consisted of 0.1％ formic acid water （A）-methanol （B） （gradient elution） at a flow rate of 1.0 mL/min. The column temperature was set at 25 ℃. The detection wavelength was set at 300 nm， and sample size was 10 μL. Using polyamide resin as material， the yield of N-（E）-p-coumaroyltyrosine as indicators， single factor test was used to optimize the purification technology of N-（E）-p-coumaroyltyrosine， such as concentration， sample size， stationary adsorption time. RESULTS： The linear range was 2.575-51.50 μg （r＝0.999 9） for N-（E）-p-coumaroyl-tyrosine. The limit of quantification was 0.000 618 μg， and the detection limit was 0.000 129 μg. RSDs of precision， stability and reproducibility tests were all lower than 3％. The recoveries were 97.04％-102.43％ （RSD＝2.06％， n＝6）. The optimal purification technology was as follows： the concentration of the sample solution was 0.04 g /mL （by the leaves of A. cantoniensis）； sample capacity 50 mL； the sample flow rate was 1.0 mL/min； the stationary adsorption time was 20 min； the eluting solvents were ammonia containing water （containing 0.1％ acetic acid）， 20％ ethanol （containing 0.1％ acetic acid） and ammonia（pH 10）. Average yield was 98.94％，average dry paste content was 61.17 mg/g， and average dry paste purity was 19.73％ by optimal purification technology. CONCLUSIONS： Established method is simple， accurate and stable. The optimized technology is stable and feasible.

Objective: To evaluate the relationship between-590C/T polymorphisms of interleukin-4 gene and periodontitis by meta-analysis. Methods: Databases including PubMed, EMbase, Web of Science, CBM, CNKI, Wanfang and VIP, were searched from their establishment to December 31, 2016. Only case-control studies on the association between -590C/ T polymorphisms of interleukin-4 gene and periodontitis were selected, and the references of those selected studies were searched by hand. Assessment and data extraction were conducted by 2 reviewers independently, and the meta-analysis was performed with the R 3.3.1 software. Results : The meta-analysis showed that there was no association between the IL-4-590C/T polymorphism and periodontitis susceptibility (T vs C: OR=1.19, 95%CI=0.81~1.76, P =0.37). Subgroup analysis showed that the IL-4 -590 TT was associated with periodontitis in Caucasians (TT vs CT: OR=1.75, 95%CI=1.10 ~ 2.78, P = 0.02), but no significant association in Asians (P =0.63). Meanwhile, IL-4 gene polymorphisms were not significantly associated with the susceptibility of chronic periodontitis (CP) or aggressive periodontitis (AgP) ( e. g. T vs C, CP: P=0.45; AgP: P =0.45). Conclusion Based on current evidence, the polymorphisms of IL-4-590C/T are associated with the increased risk of periodontitis in Caucasians.

Objective To compare the therapeutic effects between three-dimensional conformal radiotherapy (3DCRT) and intensity-modulated radiotherapy (IMRT) in patients with stage Ⅱ/Ⅲ esophageal cancer and investigate the prognostic factors.Methods Medical record of 2 132 patients with stage Ⅱ/Ⅲ esophageal cancer who underwent definitive radiotherapy with/without chemotherapy in 10 hospitals from January 2002 to December 2016 from were retrospectively analyzed.Among these patients,37.9％ of them were aged ≥ 70 years,33.9％ with neck and upper esophageal tumors and 66.1％ with middle and lower esophageal and borderline tumors.The median gross tumor volume (GTV) and lymph node gross tumor volume (GTVnd) was 41.6 cm3.Among them,32％ were stage Ⅱ] and 68％ were stage Ⅲ.A total of 723 patients received 3DCRT and 1 409 cases received IMRT.Patients received an equivalent dose in 2 Gy (EQD2) ≥ 60 Gy accounted for 86.1％,and 41.1％ of them received concurrent chemoradiotherapy.Results The median follow-up time was 60.8 months.The 1-,3-and 5-year overall survival (OS) of all patients was 73.9％,41.7％ and 32.6％,and the 1-,3-and 5-year progression-free survival (PFS) was 62.2％,37.3％ and 32％,respectively.Multivariate analysis demonstrated that age,primary tumor location,clinical stage,tumor target volume,EQD2 and concurrent chemoradiotherapy were the independent prognostic factors for OS.Age,primary tumor location,clinical stage,tumor target volume and EQD2 were the independent prognostic factors for PFS.The OS and PFS did not significantly differ among the low-risk,low-/moderate-risk,moderate-/high-risk and high-risk groups according to age≥70 years,tumor diameter＞5 cm,tumor volume ≥41.6 cm3 and stage Ⅲ (P＜0.001).After the propensity score matching (PSM) method,neither 3DCRT nor IMRT yielded significant advantages in OS or PFS (P=0.971;P=0.658).However,IMRT tended to yield survival benefits in low-risk patients (P=0.125).Conclusions Both 3DCRT and IMRT yield relatively high OS rate in patients with stage Ⅱ/Ⅲ esophageal cancer.The prognosis model established in this investigation can properly predict the survival of patients.Low-risk patients tend to obtain survival benefits from IMRT.

Objective To retrospectively analyze the effect of tumor length on the prognosis in stage Ⅱ/Ⅲ esophageal squamous cell carcinoma (ESCC) patients treated with definitive radiotherapy and to evaluate the role of tumor length in clinical stage for non-operative ESCC patients.Methods The data of 2 086 ESCC patients who were treated with definitive radiotherapy from 2002 to 2016 in 10 hospitals (3JECROG) were analyzed.The effect of tumor length on overall survival (OS) was analyzed and stratified analysis of tumor length was done in different stages of ESCC.Results The median OS and median progression-free survival (PFS) time of the whole group were 25.6 months and 18.2 months respectively.The Cox multivariate analysis showed that treatment moda,aga,alinical stage and tumor length were independent prognostic factors.The median,1-,3-,and 5-year OS were 28.9 months,77.3％,45.0％,and 36.3％ versus 21.9 months,69.9％,37.9％,and 28.1％ for patients with ≤ 5 cm and patients ＞ 5 cm respectively (P＜0.05).For stage Ⅱ patienta,abe median OS were 42.1 and 38.9 months respectively in ≤ 5 cm group and＞5 cm group (P=0.303).And for stage Ⅲ patienta,abe median OS were 23.9 and 19.3 months respectively in ≤5 cm group and＞5 cm group (P＜0.001).The median OS with N1was 24.1 and 18.4 montha,aespectively in ≤5 cm group and＞5 cm group (P＜0.001).Conclusions The tumor length was an independent prognostic factor for stage Ⅱ/Ⅲ patients treated definitive radiotherapy.The tumor length may be helpful in clinical staging of ESCa,aspecially for stage Ⅲ and N1.

Objective : To investigate the effect of ginsenoside Rg1 (GRg1) on human retinal microvascular endothelial cells (HRMECs) glycolysis by regulating pyruvate kinase M2 ( PKM2) expression. Methods : HRMECs were cultured in vitro and divided into normal control (NC) group, high glucose (HG) group, high glucose + ginsenoside Rg1 (HG + GRg1) group, high glucose + ginsenoside Rg1 + low expression PKM2 ( HG + GRg1 + si-PKM2) group, and high glucose + ginsenoside Rg1 + overexpression PKM2 (HG + GRg1 + OE⁃PKM2) group. si-PKM2 and OE⁃PKM2 were transfected into HRMECs cells by cell transfection. The expression of PKM2 mRNA in HRMECs was detected by qRT⁃PCR. The expression levels of related proteins in HRMECs were detected by Western blot. The number of lumen formation in vitro was observed under an inverted microscope to quantify the angiogenesis ability. Cell culture medium of each group was collected, and glucose intake, lactate production and adenosine triphosphate(ATP)content were detected by glucose detection kit, lactate detection kit and ATP detection kit,re spectively. Results : HG induced HRMECs significantly increased the number of blood vessel formation, glycolysis and PKM2 expression, while GRg1 treatment significantly reduced the number of blood vessel formation, glycolysis and PKM2 expression; transfection of si⁃PKM2 assisted the inhibitory effect of GRg1 on glycolysis and angiogenesis while transfection of OE⁃PKM2 interfered with the function of GRg1 . Conclusion GRg1 inhibits angiogenesis by inhibiting PKM2 to reduce glycolysis of HRMECs.

Objective To evaluate the survival and prognostic factors of esophageal cancer treated with definitive ( chemo ) radiotherapy by applying novel radiation techniques including three-dimensional conformal radiotherapy (3DCRT) or intensity-modulated radiotherapy (IMRT). Methods Clinical data of 2762 patients with non-operated esophageal squamous cell carcinoma who underwent definitive ( chemo ) radiotherapy from 2002 to 2016 in 10 hospitals were retrospectively analyzed.The prognostic factors were also identified and analyzed. Results The median follow-up time was 60. 8 months. The 1-, 2-, 3-and 5-year overall survival (OS) of all patients was 71. 4％,48. 9％,39. 3％,and 30. 9％,respectively.The 1-,2-,3-and 5-year progression-free survival (PFS) was 59.5％,41.5％,35.2％,and 30％,respectively.The median survival was 23 months.The median time to progression was 17. 2 months.Multivariate analysis demonstrated that age, primary tumor location, clinical stage, tumor target volume, EQD2 and treatment mode were the independent prognostic factors for OS.Primary tumor location,clinical stage,tumor target volume and EQD2 were the independent prognostic factors for PFS. Conclusions In this first large-scale multi-center retrospective analysis of definitive ( chemo) radiotherapy for esophageal squamous cell carcinoma in China, the 5-year OS of patients with esophageal squamous cell carcinoma is significantly improved by 3DCRT, IMRT combined with chemotherapy drugs. However, the findings remain to be validated by prospective clinical trials with high-level medical evidence.

Objective: To evaluate the prognostic factors of T1-2N0M0 esophageal squamous cell carcinoma (ESCC) treated with definitive radiotherapy. Methods: The clinical data of 196 patients with T1-2N0M0 ESCC who were treated with definitive radiotherapy in 10 hospitals were retrospectively analyzed. All sites were members of Jing-Jin-Ji Esophageal and Esophagogastric Cancer Radiotherapy Oncology Group (3JECROG). Radiochemotherapy were applied to 78 patients, while the other 118 patients received radiotherapy only. 96 patients were treated with three-dimensional conformal radiotherapy (3DCRT) and 100 treated with intensity-modulated radiotherapy (IMRT). The median dose of plan target volume(PTV) and gross target volume(GTV) were both 60 Gy. The median follow-up time was 59.2 months. Log rank test and Cox regression analysis were used for univariat and multivariate analysis, respectively. Results: The percentage of normal lung receiving at least 20 Gy (V₂₀) was (18.65±7.20)%, with average dose of (10.81±42.05) Gy. The percentage of normal heart receiving at least 30 Gy (V₃₀) was (14.21±12.28)%. The maximum dose of exposure in spinal cord was (39.65±8.13) Gy. The incidence of radiation pneumonia and radiation esophagitis were 14.80%(29/196) and 65.82%(129/196), respectively. The adverse events were mostly grade 1-2, without grade 4 toxicity. Median overall survival (OS) and progression-free survival (PFS) were 70.1 months and 62.3 months, respectively. The 1-, 3- and 5-year OS rates of all patients were 75.1%、57.4% and 53.2%, respectively. The 1-, 3- and 5-year PFS rates were 75.1%、57.4% and 53.2%, respectively. Multivariate analysis demonstrated that patients′age (HR=1.023, P=0.038) and tumor diameter (HR=1.243, P=0.028)were the independent prognostic factors for OS, while tumor volume were the independent prognostic factor for PFS. Conclusions Definitive radiotherapy is a promising therapeutic method in patients with T1-2N0M0 ESCC. Patients′ age, tumor diameter and tumor volume may impact patients′ prognosis.