A 15-year-old male patient presented with intermittent red secretion on the face, upper limbs, nail grooves, fingers and toes for more than 10 months. Ten months prior to the presentation, red secre-tions began to emerge on the face, especially ears, eyes and the surrounding area of nose, following a severe psychical stimulus. The attack typically lasted 2-3 minutes before spontaneous stop with the formation of blood crusts. The frequency was 1-4 per day during the attack of hemathidrosis which might last 1 week. The interval was irregular and varied from 10 days to months. There was no blood stasis, bruise or wound left on the skin after the wipe of bloodstain, and no itch or pain was complained of. Before the secretion of bloody sweat, the patient was always fidgety, irritable, and sometimes excited or difficult to sleep, but the secretion never occurred at night. After several episodes, bloody sweat also appeared in succession and symmetrically on the forearms, extensor crura, fingers, toes and perionychium. The cell components of bloody sweat were identical to those of peripheral blood. The patient was diagnosed as hemathidrosis. No abnormality was found in labora-tory tests, histopathological examination, electron microscopy or radiology. No relief of symptoms was achieved after treatment with vitamin C, rutoside and oryzanol. Hemathidrosis is a rare disease, and to clarify its patho-genesis is essential for its treatment.

Objective To investigate the therapeutic effects of pulsed CO2 laser irradiation in the treatment of syringoma. Methods A retrospective comparison was performed between 28 syringoma patients treated with a pulsed CO2 laser and 30 patients treated with a continuous CO2 laser. The therapeutic effects and treatment complications were compared statistically. Results The treatments were rated effective for all patients in both groups. The outcome was rated " excellent" for 86. 7% of the pulse-treated group and 78. 6% for those treated with continuous las-ing. There was no significant difference between the two groups. There was a significant difference in the rate of pigmentation and atrophic scarring. The pulsed group showed lower levels of both effects than the continuous group. Conclusion The pulsed CO2 laser is a better tool for treating syringoma in terms of safety, efficiency and side effects.

Objective To investigate the expression of CD40/CD40L on peripheral blood mononuclear cells (PBMC) in patients with recurrent genital herpes (RGH). Methods Flow cytometric analysis was employed to identify the expression of CD40 and CD40L on PBMC and the T lymphocyte subgroups in the peripheral blood of 30 patients with RGH and 20 healthy controls. Results Percentage of the cells with either CD40 or CD40L expression in the patients with RGH was significantly lower than that in the controls (P = 0.0061 and 0.041, respectively). The RGH group had a significantly lower percentages of CD3+ T cells (P=0.025) and CD4+ T cells (P = 0.032) as compared to the controls. Conclusion Reduced costimulatory interaction of CD40 and CD40L in RGH patients may be one of the important factors for the recurrence.

Objective To determine the value of ERCP and EST after Billroth gastroenterostomy. Methods ERCP was used in 31 patients after Billroth- Ⅰ gastroenterostomy, 12 of whom received EST. It was in 137 patients after Billroth-Ⅱ gastroenterostomy, of the 34 received EST and 4 EPBD.Results Billroth- Ⅰ gastroenterostomy ERCP was successfully performed in 28 out of the 31 patients and EST in 11 out of the 12 patients. Billroth- Ⅱ gastroenterostomy ERCP was successfully performed in 109 out of the 137 patients and EST in 31 out of the 38 patients. There were no serious complications in patients receiving endoscopic treatments. Concluasion The success rates of ERCP and EST are high in patients with bile duct lithiasis after Billroth-gastroenterostomy. Endoscopic treatment or cholangioduodenostomy has good therapeutic effects.

Objective To evaluate the performance verification of AFP and CEA detected by Roche Cobas 6000 automatic electrochemical luminescence analyzer.Methods Serum samples in hospital patients during April 1 to 4,2014 were collected and mixed.Samples with the different levels of AFP and CEA were prepared.Referring to CLSI EP files and other docu-ments,the precision,accuracy and the reported linear range of AFP and CEA from Roche Cobas 6000 automatic electrochem-ical luminescence analyzer were verified.Results AFP and CEA in the quality control sera with low and high values were detected by Roche Cobas 6000 automatic electrochemical luminescence analyzer.The coefficient of variations (CV)of inter batch precision were AFP (6.53%,8.38%),CEA (8.15%,7.84%),and the CV of within batch precision were AFP (3.97%,6.51%),CEA (4.77%,4.52%),respectively.10 copies of laboratory quality control were analyzed,issued by the National Center for clinical laboratory in 2013.The largest bias between detection results and“target value”were AFP (-12.62%)and CEA (-10.71%)respectively,which were all within the measurement range of quality assessment.Analysis of measuring range in the linear range experiment were AFP (0.5~1 000 IU/ml),CEA (0.2~1 000 ng/ml),and the clini-cal reportable range were AFP (0.5~1 000 IU/ml),CEA (0.2~1 000 ng/ml),respectively.Conclusion Detection of AFP and CEA from Roche Cobas 6000 automatic electrochemical luminescence analyzer had a good performance.Meanwhile,the Roche Cobas 6000 automatic electrochemical luminescence analyzer could meet the clinical routine testing requirements.

Objective To investigate the changes and significance of ubiquitination of Foxp3 pro-tein during the acute phage of Kawasaki disease ( KD) .Methods Forty-eight children with KD and twenty-eight age-matched healthy children were recruited in this study.Co-immunoprecipitation and Western blot assays were performed to determine the poly-ubiquitination status of Foxp3 in CD4+T cells.The percentages of CD4+CD25high Foxp3+regulatory T cells ( Treg) and the levels of IL-10, transforming growth factor-β( TGF-β) , cytotoxic T lymphocyte-associated protein-4 ( CTLA4 ) , STIP1 homology and U-Box containing protein 1 (STUB1), heat shock protein 70 (HSP70), ubiquitin-specific-processing protease 7 (USP7) and pSTAT3 were analyzed by flow cytometry analysis.Quantitative real-time PCR was used to evaluate the tran-scription levels of TLR1-10, IL-1R1, IL-1RAP, IL-6Rα, gp130, TNFR1, MyD88 and RIP1 in CD4+T cells.Plasma concentrations of IL-1β, IL-6 and TNF-αwere measured by enzyme-linked immunosorbent as-say.Results (1) The percentages of Treg cells and the levels of IL-10, TGF-β, CTLA4 and forkhead box P3 (Foxp3) in patients with acute KD were lower than those of healthy subjects (P<0.05), while the poly-ubiquitination of Foxp3 protein was significantly enhanced in patients with acute KD [(0.52 ±0.19) vs (0.08±0.02),P<0.05].Meanwhile, the four former items in KD patients with coronary artery lesions (KD-CAL+) were lower than those of KD patients without coronary artery lesions (KD-CAL-) (P<0.05), while the polyubiquitination level of Foxp3 protein in KD-CAL+group was much higher than that of KD-CAL-group [(0.70±0.28) vs (0.43±0.17), P<0.05].The levels of Treg cells, IL-10, TGF-βand CTLA4 in patients with KD were increased and the ubiquitination of Foxp3 protein was inhibited [(0.24±0.10) vs (0.52±0.19), P<0.05] upon the treatment with IVIG.(2) The levels of STUB1 and HSP70 in CD4+T cells were significantly elevated during acute KD, while the levels of USP7 were decreased (P<0.05).The ratios of STUB1/USP7 in patients with acute KD were much higher than those of the control group [(2.65± 0.92) vs (1.09±0.37), P<0.05], but were significantly decreased after IVIG therapy [(1.46±0.53) vs (2.65±0.92), P<0.05].A negative correlation was found between STUB1/USP7 ratio and Foxp3 level during acute KD (r=-0.56, P<0.05).Moreover, KD patients with CAL+showed higher levels of STUB1 and HSP70 and higher ratios of STUB1/USP7 (P<0.05), but lower levels of USP7 as compared with those of KD-CAL-group (P<0.05).(3) The plasma concentrations of inflammatory cytokines (IL-1β, IL-6 and TNF-α), the levels of surface receptors ( IL-1R1/IL-1RAP/TLR4, IL-6Rα/gp130 and TNFR1) and its downstream molecules ( MyD88, pSTAT3 and RIP1) in CD4+T cells were up-regulated during acute KD ( P<0.05), especially in patients with CAL+, but were down-regulated upon the IVIG therapy (P<0.05).No significant differences with other TLRs were found among the groups (P>0.05).Conclusion Hyper-ubiq-uitination of Foxp3 protein might be involved in the immune dysfunction during Kawasaki disease.

Objective To investigate the role of IL-35-producing regulatory B cells(IL-35 + Breg)in immunological pathogenesis of Kawasaki disease (KD).Methods Thirty-two children with KD and 28 age-matched healthy children were allowed to participate in this study.Flow cytometry was performed to evaluate the proportions of IL-35 + Breg as well as requlatory T cells (Treg)and expression levels of associated molecules such as programmed death-ligand 1 (PD-LI),CD169,programmed death 1 (PD-1),CD43,IL-12p35,epstein-Barr virus induced 3 (IL-27 EBI3).IL-12 receptor beta 2 (IL-12 Rβ2),IL-27 receptor alpha (IL-27 Rα),phosphated signal transducer and activator of transcription 1 (pSTAT1) and phosphated signal transducer and activator of transcription 3 (pSTAT3).Transcription levels of the Src homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2),phosphatase and tensin homolog (PTEN),vav1 guanine nucleotide exchange factor(Vav) in CD19 + B cells were determined by using quantitative real-time PCR.Plasma concentrations of IL-35,tumor necrosis factor α(TNF-α) and IL-12 were measured by adopting enzyme-linked immunosorbent assay.Results (1) The proportions of IL-35 +Breg and its expressions of IL-12p35,IL-27EBI3 and IL-10 in patients with acute KD were lower than those of healthy controls [IL-35 + Breg:(5.79 ± 2.60) ％ vs (12.65 ± 5.34) ％;F =19.23,9.70,14.30.7.08;all P ＜ 0.05],but they were significantly increased after intravenous immune globulin (IVIG) treatment [IL-35 + Breg:(10.52 ± 4.95) ％;all P ＜ 0.05].(2) The proportions of Treg and its transcriptional levels of IL-12p35 and IL-27 EBI3 were down-regulated during acute KD [Treg:(4.12 ± 1.51) ％ vs (8.06 ± 3.32) ％;F =19.70,17.69,38.22;all P ＜ 0.05],but were increased after therapy [Treg:(7.39 ± 2.85) ％;P ＜ 0.05].A positive correlation was found between the proportions of Treg and IL-35 + Breg during acute KD (r =0.69,P ＜ 0.05).Meanwhile,plasma concentrations of IL-35 and expression levels of IL-12Rβ2,IL-27Rα,pSTAT1 and pSTAT3 in CD19 + B cells were significantly down-regulated in children with acute KD,but they were increased after treatment(F =8.09,7.54,7.69,5.89,12.59,all P ＜ 0.05).(3) Compared with healthy controls,expressions of PD-L1 and CD169 on CD14 + cells and plasma concentrations of TNF-α and IL-12 were elevated during acute KD (F =24.94,16.53,34.71,19.51;all P ＜ 0.05).Expression levels of PD-1,CD43 and its downstream molecules (SHP-2,PTEN,Vav) in CD19 + B cells were down-regulated during acute KD (F =6.43,5.57,19.52,10.37,11.37;all P ＜ 0.05),and restored remarkably after therapy (all P ＜ 0.05).Conclusion Insufficiency of IL-35 + Breg and its expression of IL-35 may be the important factors contributing to immunological dysfunction in KD.

Objective To investigate the changes and significances of inducible IL-35-producing regulatory T cells(iTR35) in immunological pathogcnesis of Kawasaki disease (KD).Methods Forty-eight children with KD and 32 age-matched healthy children (healthy control group) consented to participate in this study.Flow cytometry was performed to evaluate the proportions of CD4+ FOXP3-IL-12p35+IL-27EBI3+iTR35 and CD4+CD25high FOXP3+regulatory T cells (Treg),and expression levels of associated molecules such as programmed death-ligand 1 (PD-L1),CD169,programmed death 1 (PD-1),CD43,IL-12p35,Epstein-Barr virus induced 3 (IL-27EBI3),glycoprotein 130(gp130),IL-12 receptor beta 2 (IL-12Rβ2),phosphated signal transducer and activator of transcription 1 (pSTAT1) and phosphated signal transducer and activator of transcription 4 (pSTAT4).Transcription levels of the Sre homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2),phosphatase and tensin homolog (PTEN),Vavl guanine nucleotide exchange factor(Vav) in CD4+T cells were determined by quantitative real-time PCR.Plasma concentrations of IL-35,IL-10,TNF-α and IL-12 were measured by enzyme-linked immunosorbent assay.Results (1) The proportions of iTR35 and its expressions of IL-12p35 and IL-27EBI3 in patients with acute KD dccreased remarkably[iTR35:(0.72±0.26) ‰ vs (1.65±0.43) ‰,P＜0.05],and restored after treatment [iTR35:(1.58±0.63) ‰ vs (0.72±0.26) ‰,P＜0.05].(2) The proportions of Treg and transcriptional levels of IL-12p35 and IL-27EBI3 were down-regulated during acute phase of KD [Treg:(3.26±1.21) ％ vs (7.26±2.86) ％,P＜0.05],and increased to some extent after therapy [Treg:(5.89±2.60)％ vs (3.26±1.21)％,P＜0.05].Meanwhile,plasma concentrations of IL-35 and IL-10,and expressions of gp130,IL-12Rβ2,pSTAT1 and pSTAT4 in iTR35 of patients with acute KD were found lower than those of the healthy control group (all P＜0.05),and increased after treatment (P＜0.05).Additionally,positive correlations were found between plasma concentrations of IL-35 and the proportion of iTR35 or its expressions of IL-12p35 and IL-27EBI3,respectively.(3) Expressions of PD-L1 and CD169 on CD14 + cells and plasma concentrations of TNF-α and IL-12 were elevated significantly during acute KD(all P＜0.05),as well as expression levels of the ligands (PD-1 and CD43) and its downstream molecules (SHP-2,PTEN,Vav) in CD4 + T cells were found to be lower in patients with acute KD (P＜0.05),and restored remarkably after therapy.Conclusion Insufficiency of iTR35 and its expression of IL-35 might be one of the important factors contributing to immunological dysfunction in KD.

Aim With metabolomics method, to study Shen-Mai decoction’ s function on protecting the myo-cardial injured rats caused by doxorubicin for probing into the functioning mechanism of Shen-Mai decoction’ s medical effect. Methods By means of UPLC-TOF-MS, the metabolites of urine of the rats treated by Shen-Mai decoction were analyzed. Then, the differ-ences between each group of the metabolites were sought with PLS-DA ( the partial least square discrimi-nant analysis ) and OPLS-DA ( the orthogonal partial least squares discriminant analysis ) . VIP ( variable importance in projection ) and t test were used to screen out potential biomarkers. Results Fourteen endogenous metabolites such as succinyladenosine, a-denosine 2′, 3′-cyclic phosphate, S-( 3-methylbu-tanoyl )-dihydrolipoamide-E, cis-4-hydroxycyclohexy-lacetic acid, phenylbutyrylglutamine, 3-butyn-1-al, 3-hydroxytetradecanedioic acid, dihydrolipoamide and pyruvic acid, etc. were characterized. Conclusions The results indicate that Shen-Mai decoction can pro-tect the body from myocardial injury by regulating pu-rine metabolism, some acid metabolism, fat metabo-lism and energy metabolism, etc. The study expounds the functioning mechanism for Shen-Mai decoction ’ s medical effect in the body and provides theoretical grounds for the rationality of the two medical herbs ’ compatibility and their combination in clinical treat-ment of diseases.

Aim To study the effect of emodin in Po-lygonum multiflorum on the expression of CYP450 isoenzymes in L02 hepatocytes and explore its mecha-nism of cytotoxicity. Methods L02 cells were treated with different concentrations of emodin. Cell viability was examined by MTS assay kit, and cell membrane injury was examined by detecting the release rate of lactate dehydrogenase( LDH) . The expression of cyto-chrome P450 mRNA was detected by real time PCR. Results The result of MTS assay showed that L02 cells viability was significantly reduced following expo-sure to emodin in a concentration and time dependent manner. The LDH release rate of L02 cells significant-ly increased after exposure to emodin for 48 h com-pared with the control group. On the mRNA level, compared with the control group,emodin had inductive effects on mRNA of each CYP450 enzyme, while had significant inductive effects on mRNA of CYP1 A1 and CYP1 B1 in a concentration and time dependent man-ner. Conclusion Emodin in Polygonum multiflorum may generate significant liver injury in L02 cells and has inductive effects on CYP450 enzyme activity.