Objective To investigate the levels of cytokines in sera of patients with various types of psoriasis.Methods Six cytokines, sIL-2R、 IL-2、 -4、 -10、 -12 and IFN-γ , were detected by sandwich ELISA in the sera from 15 patients with guttate psoriasis, 23 plaque psoriasis, 9 pustular psoriasis, 9 arthropathic psoriasis and 9 erythrodermic psoriasis.Results Significantly higher levels of cytokines were observed in guttate psoriasis for sIL-2R (P<0.01), in plaque psoriasis for IL-4,-12 and sIL-2R (P< 0.05 or < 0.01), in pustular psoriasis for IL-4 and -10 (P< 0.05), in arthropathic psoriasis for IL-10 (P< 0.01), in comparison with the controls.The levels of the six cytokines were increased in erythrodermic psoriasis with no significant difference from the controls.The ratio of the average level of IL-4 to IFN-γ was 1.57 in pustular psoriasis, 0.61 in plaque psoriasis, 0.30 in gutatte psoriasis, 0.24 in erythrodermic psoriasis, and 0.02 in arthropathic psoriasis.Conclusion There is different expression of cytokines in sera of patients with various types of psoriasis.

Objective To establish a polymerase chain reaction(PCR) method to amplify DNA of M.leprae from fixed and paraffin－ embedded tissue(FPET). Methods The DNA of M.leprae was released from FPET by using Texpat Kit and purified with 100％ alcohol. The primers RPOT(1) and RPUT(2) were used to conduct the PCR. Results A total of 32 samples were examined. Out of 32 samples with BI of more than 1＋ , 28 were positive for PCR. The PCR was negative in a sample with BI=0. The sensitivity of PCR reached a level of 0.04 pg DNA. Conclusion This PCR method is very useful for amplifying the DNA of M.leprae from FPET.

Objective To explore whether there were differences in genotypes between strains of M.leprae in China or not. Methods Polymerase chain reaction (PCR) was used to detect the genotypes of M.leprae in samples from patients with leprosy in different parts of China. Results Out of 16 samples from 9 provinces, 91 bp DNA fragments were present in 7 samples and 97 bp DNA fragments in 9 samples. Conclusion Above- mentioned results suggest the genotyping differences in strains of M.leprae with different distribution in areas of China.

Objective To investigate the levels of cytokines in sera of patien ts with various types of psoriasis. Methods Six cytokines, sIL－2R、 IL－2、－ 4、－10、－12 and IFN－? , were detected by sandwich ELISA in the sera from 15 patients with guttate psoriasis, 23 plaque psoriasis, 9 pustular psoriasis, 9 arthropathic psoriasis and 9 erythrodermic psoriasis. Results Significantly hig her levels of cytokines were observed in guttate psoriasis for sIL－2R (P

Objective To genotype the clinically isolated strains of M.leprae by genome sequencing analysis. Methods PCR amplification was used to produce the 200 bp partial rpoT gene fragments from 2 standard strains and the isolated strains of M. leprae isolated from clinical specimens in 7 areas of China. The fragments were sequenced by BigDye Terminator Cycle Sequencing Reaction kit. Results① 12 rpoT－ 91/97bp DNA fragments and 2 rpoT－ 194/200 bp DNA fragments were obtained from 13 clinically isolated strains of M. leprae by PCR amplification;② based upon genome sequencing analysis, a component of 3－ copy or 4－ copy“ GACATC” repeat sequence was found in the nucleotide sequence of rpoT－ 194/200 bp DNA fragment. Conclusion The genome sequencing analysis can be used to objectively and accurately genotype M.leprae, and it is a useful tool for epidemiological study on transmission and infection of leprosy. However, the longer DNA fragments are necessary when sequencing analysis is conducted. Therefore this method needs further improvement because only the shorter DNA fragments amplified from paraffin－ embedded tissue.

Objective To compare the performance of 2％ (w/v) agarose gel,2％ (w/v) Metaphor agarose gel and 10％(w/v) nondenaturating polyacrylamide gel in the PCR-restriction fragment length polymorphism (RFLP) analysis of mycobacterial heat shock protein 65 (hsp65) gene.Methods This study included 8 Mycobacteria strains,including clinical isolates and standard strains of Mycobacteria tuberculosis and Mycobacterium intracellulare.Bacterialsuspension of these strains was prepared with the concentration of bacterial cells varying from 10 to 106per milliliter.PCR was performed to amplify the hsp65 gene with a pair of universal primers followed by the digestion of amplicons with two restriction endonucleases,BstE Ⅱ and Hae Ⅲ.Then,the restriction enzyme-digested fragments were subjected to electrophoresis in 2％ agarose gel,2％ Metaphor agarose gel and 10％ nondenaturating polyacrylamide gel respectively.Results As analysis of variance showed,the three gel electrophoresis methods were statistically different in sensitivity for the RFLP analysis of mycobacterial hsp65 gene (F =36.379,P ＜ 0.01).Least significance difference (LSD) procedure revealed that the 2％ agarose gel-based electrophoresis was less sensitive than the 2％ Metaphor agarose gel-and 10％ non-denaturing polyacrylamide gel-based electrophoresis (both P ＜ 0.01),and no significant differences were observed between the 2％ Metaphor agarose gel-and 10％ non-denaturing polyacrylamide gel-based electrophoresis (P ＞ 0.05).Conclusion The 2％ Metaphor agarose gel-and 10％nondenaturating polyacrylamide gel-based electrophoresis methods appear to be more sensitive than the 2％ agarose gel-based electrophoresis method for the PCR-RFLP analysis of mycobacterial hsp65 gene.

Objective.So far,it has not been established a satisfactory method for early diagnosis and studying on epidemiology for leprosy,we want to develop a molecular biological method for solving this point.Materials and methods.Based on the M.leprae gene coding groEL,65kD and 16S rRNA,three polymerase chain reactions were developed by using plikaytis',Woods' and Pattyn's procedures.It was optimized that the experimental parameters for each PCR,and a comparative study on practivity among three PCRs was also conducted for practical purpose.Results and conclusion.For detecting infection with M.leprae,all of PCRs established by us were highly sensitive and specific,but for practicl purpose,the woods'PCR optimized by us ought to be chosen firstly.

1), attributable risk (AR)=13.33%; ⑤Even though the serum specimens were taken from PB patients, the Ab of majouity of patients converted to positive when relapse occurred. In the majority of patients relapsed the levels of IgM-AbL tended to be increasing or pesistently positive. Usually relapse occurred 1-2 years after IgM-AbL was converted to positive. Relapse occurred 11-30 years after the patients were cured. In very rare case downgrading(from tuberculous to borderline leprosy) occurred. ⑥The levels of IgM-AbL gradually decreased in all relapsed patients after effective treatment except one case whose IgM-AbL was persistently positive. Conclusions The above results indicate that the ND-ELISA might be useful in screening early M.leprae infection and in predicting and monitoring the relapse of leprosy, especially in multibacillary patients.

Objective To test the drug susceptibility of 16 Mycobactena tuberculosis isolates from patients with cutaneous tuberculosis,and to provide a basis for the treatment of this entity.Methods Sixteen strains of mycobacterium were isolated from patients with cutaneous tuberculosis.and they were consistently identified as M.tuberculosis by biochemistry and molecular biology.Absolute concentration method was used to test the susceptibility of these isolates to isoniazid.streptomycin.rifampicin and ethambutol.For two strains resistant to streptomycin,PCR and sequencing were performed to analyse the mutation of rpsL gene.Results Out ofthe 16 strains of M tuberculosis.2 were resistant to streptomycin but sensitive to isoniazid,rifampicin and ethambutol,and 14 were sensitive to isoniazid,streptomycin,rifampicin and ethambutol.Sequencing of the rpsL gene revealed a mutation of AAG to AGG at codon 43 in one streptomycinresistant strain and a substitution of CGC bv CAC at codon 54 in another resistant strain.Conclusions In past five years,the resistance ratio of M tuberculosis was low in patients with cutaneous tuberculosis,and streptomycin resistance predominated in these strains.

Objective To study the effect and mechanism of dexmedetomidine combined with sufentanil on postoperative analgesia in elderly patients with abdominal surgery. Methods Ninety-six elderly patients having undergone abdominal surgery in the First Affiliated Hospital of Wenzhou Medical University from January 2016 to June 2017 were enrolled, and they were divided into a control group and an observation group by random number table method, 48 cases in each group. General anesthesia was performed in the operation, and after surgery venous analgesic pump was used for analgesia in both groups. Analgesic method: the control group was given sufentanil 2 μg/kg and tropisetron 5 mg; the observation group was given dexmedetomidine 2 μg/kg, sufentanil 2 μg/kg and tropisetron 5 mg. The changes of pain and sedation score, conventional indexes [systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), respiratory rate (RR), pulse blood oxygen saturation (SpO2)], oxidative damage indexes [lipid peroxide (LPO), glutathione peroxidase (GSH-Px), Cu-Zn superoxide dismutase (Cu-Zn SOD)], stress response indexes [cortisol (Cor), epinephrine, norepinephrine (NE)], platelet activation indexes granule membrane protein-140 (GMP-140), thromboxane A2(TXA2) and the incidence of adverse reactions were observed in both groups. Results ① After surgery the visual simulation score (VAS) and Ramsay score in both groups were higher than those before surgery, and showed a tendency firstly increased and then decreased, and reached to peak value 2 hours after operation [VAS score:the control group was 3.24±0.98 vs. 1.95±0.93, observation group was 3.19±1.03 vs. 1.98±0.95; Ramsay score:the control group was 3.26±0.51 vs. 1.90±0.45, observation group was 3.77±0.53 vs. 1.92±0.42], began to decline 6 hours after operation, reached to valley value 48 hours after operation, and there was no significant difference in VAS scores between the two groups (2.02±0.64 vs. 1.98±0.95), Ramsay score was significantly higher in observation group than that in control group (2.59±0.41 vs. 2.10±0.21). ② Since 2 hours after the operation, the SBP, DBP, HR and RR in the observation group began to be lower than those in control group, 12 hours after surgery their values reached their valleys [SBP (mmHg, 1 mm Hg = 0.133 kPa): 113.2±13.5 vs. 122.1±10.3, DBP (mmHg): 67.5±9.9 vs. 76.4±8.6, HR (bpm): 64.5±6.9 vs. 71.4±7.5, RR (bpm): 14.8±1.1 vs. 15.8±0.8, all P < 0.05]; while SpO2did not change a great deal. ③ LPO, Cor, epinephrine, NE, GMP-140, TXA2of two groups after operation were higher than those before operation, GSH-Px and Cu-Zn SOD were lower than those before operation. However, LPO, Cor, epinephrine, NE, GMP-140, TXA2in observation group were significantly lower than those in control group [LPO (μmol/L): 6.4±0.8 vs. 9.8±1.1, Cor (ng/L): 148.2±19.1 vs. 239.3±27.8, epinephrine (μg/L): 124.2±13.9 vs. 207.1±23.5, NE (μg/L): 109.2±14.8 vs. 183.3±21.6, GMP-140 (μg/L): 27.13±3.82 vs. 39.06±4.83, TXA2(ng/L): 422.30±53.74 vs. 610.43±73.21, all P < 0.05], GSH-Px and Cu-Zn SOD in observation group were significantly higher than those in the control group [GSH-Px (U/L): 426.7±58.7 vs. 307.9±51.2, Cu-ZnSOD (μg/L): 311.3±42.5 vs. 231.6±34.1, all P < 0.05].④ The incidence of adverse reaction nausea in the observation group was significantly lower than that in the control group [4.17% (2/48) vs. 37.5% (18/48)]. Conclusion Dexmedetomidine combined with sufentanil used in elderly patients after abdominal surgery has significant analgesic effect, can effectively inhibit platelet activation, and decrease the contents of GMP-140 and TXA2.