Objective: To establish a rapid approach to the detection and identification of Mycobacteria from lesions of patients with suspected Mycobacterial infections. Methods: Specimens were obtained from five patients suspected to have Mycobacterial infections. DNA extracted from clinical samples was amplified by nested PCR. The PCR products were digested with Hha Ⅰ, MboⅠ, and BstUⅠ restriction enzymes and applied to PAGE. The species of Mycobacteria were determined by restriction fragment length polymorphism (RFLP) analysis. Identification of Mycobacteria culture was also performed in 3 patients. Results: M. marinum was found in two patients diagnosed as swimming pool granuloma. M. tuberculosis was found in one patient diagnosed as infectious skin granuloma. All these 3 Mycobacteria were confirmed by Mycobacteria culture. A strain of M. tuberculosis and a strain of M. fortuitum were detected in remain two patients. Conclusion: The results above indicate that PCR-RFLP analysis is rapid and reliable in detection and identification of different Mycobacteria species from skin tissues. Application of this method will be helpful for early diagnosis and treatment of Mycobacteria skin infections.

Objective To investigate the possible role of mycobacterial infection in the pathogenesis of sarcoidosis.Methods DNA fragments of mycobacterial16S rRNA gene and the complex-specific insertion sequence IS6110of M.tuberculosis were detected by polymerase chain reaction(PCR)in formalin-fixed,paraffin-embeded skin biopsy specimens from12patients with sarcoidosis.Results Mycobacterial common sequence16S rRNA gene was found in6patients,among which5were further confirmed as M.tuberculosis,the other one was atypical mycobacteria.The IS6110DNA fragment amplified by PCR was confirmed by DNA sequencing.Conclusion The above findings suggest an aetiological role of mycobacterial infection,especially M.tuberculosis,in some patients with sarcoidosis.

Objective To investigate the expression of extracellular signal-regulated kinase 1/2(ERK1/2) and transforming growth factor ?1(TGF-?1) in heart of spontaneous hypertensive rat.Methods The expression of ERK1/2 and TGF-?1 was determined by immunohistochemistry and Western blot.Results In arteriole of the heart of 8,16,and 20 week-old SHR,the rate of positive ERK1/2 staining at the endothelium(15.38%,76.97% and 72.72%,respectively) and at the vascular smooth muscle cell(VSMC) of 16 and 20 week-old SHR(5.49% and 6.83%,respectively)were significantly higher than that of control group(P

Objective To investigate the genetic polymorphism of HPA-1 to-16 antigens in Zhejiang Han population.Methods A total of 120 samples from unrelated donors of Zhejiang were genotyped by Polymerase Chain Reaction with Sequence-Specific Primers(PCR-SSP).Results The allele frequencies of HPA-1b,-2b,-3b,-6bw,-15b were 0.0125,0.0542,0.3833,0.0125 and 0.4833,respectively.HPA-4b,-5b,-7bw,-8bw,-9bw,-10bw,-11bw,-12bw,-13bw,-14bw and-16bw alleles were not detected.The mismatch probability(MP) of HPA-3 and 15 were 0.36 and 0.37,respectively,in raneom platelet transfusion.Conclusion There is no difference of HPA polymorphism distribution between Han population in Zhejiang and those in other areas of China.The polymorphism of HPA-3 and-15 are more abundant than other HPA antigens,so they may cause alloimmunity in random platelet transfusion recipients and between mothers and fetuses.

Objective To explore the clinical feature and treatment method of the dopa responsive dystonia (DRD).Methods To observe the clinical manifestation,auxiliary test and the response on the treatment of levodopa in 6 patients from 4 families.Results 1 patient for childhood onset and 2 patients for adolescent onset with DRD, first symptom showed leg dystonia and toe walking with difficulty, whereas 3 patients of adult onset showed tremor and rigidity. Babinski sign was presented in 3 cases. Diurnal fluctuation in symptom severity occurred in all cases. All patients had obvious response to small dose levodopa in 1～6 days. In 2 cases, the effect was maintained for 7 years, without increasing the intake of levodopa.Conclusion The clinical feature of DRD was more remarkable,the therapeutic effectiveness of levodopa was rapid,sustained and obvious.

Objective To investigate the effects of comprehensive pain management on pains and recovery of knee function of patients who hadundergone total knee arthroplasty(TKA).Methods A total of 50 patients who had undergone single-knee TKA surgery were selected and divided into 2 groups of 25 patients each:the study group and the control group.The control group was given patient-controlled epidural analgesia while the study group was given comprehensive pain management.The two groups were compared in terms of score on pain within 7 days after operation,knee joint activity and the incidence of adverse reaction within 10 days after operation(the 1st,3rd,7th,10th day).Results Within 7 days,the scores on pain in the study were significantly lower than those in the control group(P<0.05).The scores on the knee joint activity in the study group on the 1st,3rd,7th,10th day were significantly higher than those in the control group(P<0.05)and the incidences of adverse reaction were significantly lower as well (P<0.05).Conclusion Comprehensive pain management is effective for post-surgery pain control and the hastened TKA patients, and the improved quality of life.

Lipase from Burkholderia cepacia complex is one of the most versatile biocatalyst and is used widely in many biotechnological application fields including detergent additives, the resolution of racemic compounds, etc. Based on the known whole genomic information of B. cepacia, both ampicillin and kanamycin were added to the TB-T media, the traditional selective media, to screen B. cepacia complex strains from rhizosphere soil samples. The single colonies on the plates with the modified TB-T media were then qualitatively determined the ability to produced the extracellular lipase in the rhodamine B-olive oil agar plates. Thirty-five strains of lipolytic pseudo-B. cepacia complex were isolated and the positive rate of lipolytic bacteria was 65%. Among them, 15 pseudo-B. cepacia complex strains with the tolerance to benzene, n-hexane and n-heptane at the concentration of 10% (V/V) were selected and identified by the recA gene sequence. All of the 15 lipolytic bacteria belonged to the B. cepacia complex.

Objective To observe the expressions of antimicrobial peptides LL-37,human beta defensin2 (HBD-2) and HBD-3 in the lesions of lupus vulgaris and tuberculosis verrucosa cutis,and to speculate the pathogenesis of cutaneous tuberculosis.Methods Tissue specimens were collected from the lesions of 18 patients with lupus vulgaris or tuberculosis verrucosa cutis and 15 patients with psoriasis,as well as from the normal skin of 10 healthy controls.The specimens were embedded by paraffin and subjected to an immunohistochemical analysis for the detection of antimicrobial peptides LL-37,HBD-2 and HBD-3.Statistical analysis was performed by using the SPSS 13.0 software package,and t test was carried out to compare the expressions of LL-37,HBD-2 and HBD-3 between these groups.Results LL-37 and HBD-2 were observed mainly in the upper and middle layer of the epidermis,appendages and vascular endothelial cells in cutaneous tuberculosis lesions.The expression levels of LL-37 and HBD-2 were significantly higher in tuberculosis lesions than in the normal skin (t =2.632,2.399,both P ＜ 0.05).Psoriatic and tuberculosis lesions shared a similar expression pattern for LL-37 and HBD-2.HBD-3 was absent in the lesions of cutaneous tuberculosis,but present in both psoriatic lesions and normal skin.Conclusions Both LL-37 and HBD-2 may participate in the immune response in cutaneous tuberculosis,while the absence of HBD-3 may contribute to the pathogenesis of cutaneous tuberculosis.

Objective To observe the effect of different methods such as combined treatment of traditional Chinese and western medi-cine, the simple excision and expanded resection for the treatment of granulomatous lobular mastitis. Methods 60 cases of patients treated in our hospital from Jan. 2010 to Jan. 2013 were enrolled in this study, and they were confirmed the diagnosis of granulomatous lobular mas-titis by pathology examination. According to randomly number table, they were divided into three groups and they were given the treatment of combined treatment of traditional Chinese medicine and Western medicine, the simple excision, and expanded resection respectively. Where-after, the therapeutic effect of the three groups were observed. Results There were 9 cases (45. 0%) were of cure and 11 cases (55. 0%) of recurrence in the combined treatment group. There were 16 cases (80. 0%) of cure and 4 cases (20. 0%) of recurrence in the simple ex-cision group. There were 8 cases (90. 0%) of cure and 2 cases (10. 0%) of recurrence in the combined treatment group. It shows that the recurrence rate of combined treatment group was higher than the other two groups (P<0. 05). The satisfactory rate of breast appearance after treatment were 17 (85%), 15 (75%) and 16 (80%) respctively, and there is no significant difference (P>0. 05). Conclusion The effect of extended resectionin for the treatment of granulomatous is better and it has important clinical value.

Objective To observe the expression of Toll-like receptors (TLRs) 2,4 and 9 in lesions of lupus vulgaris and tuberculosis verrucosa cutis,and to evaluate the possible roles of these receptors in the pathogenesis of cutaneous tuberculosis.Methods Skin specimens were obtained from the lesions of 18 patients with clinically and pathologically diagnosed lupus vulgaris or tuberculosis verrucosa cutis,15 patients with plaque-type psoriasis (positive control),and from perilesional normal skin of 10 patients with pigmented nevi (negative control).Immunohistochemistry was used to detect the expression of TLRs 2,4 and 9 in these specimens,and the expression intensity was expressed as absorbence values.Data were analysed by using the SPSS 13.0 software,and t test was conducted to assess the differences between the three groups of specimens.Results TLR-2 was observed mainly in the middle and upper layer of epidermis in lesions of cutaneous tuberculosis and psoriasis,and in the whole epidermis except the basal layer in normal skin,with no significant difference in the expression intensity between the three groups of specimens (0.25 ± 0.04 vs.0.25 ± 0.05 vs.0.28 ± 0.03,P ＞ 0.05).TLR-4 was weakly expressed in cutaneous tuberculosis lesions,absent or slightly expressed in normal skin and psoriatic lesions,and the expression level of TLR-4 was significantly higher in cutaneous tuberculosis lesions than in normal skin and psoriatic lesions (0.16 ± 0.07 vs.0.07 ± 0.09 and 0.02± 0.05,t =2.58,6.24,P ＜ 0.01 and 0.05).TLR-9 was expressed in the whole epidermal layer,including appendages and vascular walls,of cutaneous tuberculosis and normal skin,with a significant increase in cutaneous tuberculosis than in the normal skin (0.25 ± 0.05 vs.0.19 ± 0.05,t =2.88,P＜ 0.05).Conclusions TLRs 2,4 and 9 are all expressed in cutaneous tuberculosis losious,and the expression intensity of TLR-4 and TLR-9 is higher in cutaneous tuberculosis lesions than in normal skin,hinting that TLR-4 and TLR-9 play a certain role in the immune response of cutaneous tuberculosis.