Objective To investigate the role of nentrophils in the pathogenesis of psoriasis vulgaris. Methods Neutrophils were isolated from venous blood samples of 25 patients with psoriasis vulgaris (including 13 cases of active psoriasis and 12 cases of inactive psoriasis) as well as 25 normal human con-trols, and cultured. Then, these neutrophils were grouped and treated with lipopolysaccharide (LPS, 100 g/L),CpG-A (50 mg/L), CpG-B (50 mg/L), and RPMI 1640 culture medium, respectively, for 24 hours followed by the collection of culture supematants. Human keratinocytes (HaCaT) were cultured in the presence of su-pematants of treated or untreated nentrophils for 72 hours followed by the detection of cell proliferation with MTT assay. To determine the role of proinflammatory factors, SOD/CAT and monoclonal antibody to IL-8 and TNF-alpha of 400 u/mL were used to pretreat HaCaT cells 1 hour prior to the stimulation with super-natants of neutrophils. Results Compared with culture medium, the supematant of unstimulated neutrophils from normal controls or patients with inactive psoriasis had no significant effect on the proliferation of HaCaT cells (P ＞ 0.05), but that from patients with active psoriasis markedly promoted the proliferation of HaCaT cells (t = 2.41, P < 0.05). ARe, stimulation by LPS, CpG-A and CpG-B, the supematant of active patient-derived neutrophils significantly promoted the proliferation of HaCaT cells compared with that of normal control-derived nentrophils (t = 3.11, 2.89, 2.29, respectively, all P < 0.05). In comparison with tmstimulated neutrophils, the supematant from LPS- and CpG-A stimulated nentrophiles significantly accelerated the pro-liferation of HaCaT cells. Furthermore, the proliferation of HaCaT cells induced by the supematants of LPS-,CpG-A-, CpG-B-stimulated neutrophils from psoriatic patients was statistically suppressed by the pretreat-ment with the monoclonal antibody to IL-8, TNF-alpha and SOD/CAT (all P < 0.05). Conclusions In patients with psoriasis vulgaris, there is an abnormal secretion of IL-8, TNF-alpha and superoxide by neutrophils in peripheral blood, and these proinflammatory factors could promote the proliferation of HaCaT cells.

Objective： The current study was designed to evaluate the treatment results of nasopharyngeal carcinoma by conformal radiotherapy. Methods： From September 1997 to February 2000, a total of 41 cases of recurrent nasopharyngeal carcinoma were treated by conformal radiotherapy. Among them, 27 cases were treated by late-course hypofraction conformal radiotherapy with the dose of 20- 24 Gy at 80％ isodose, 4- 7 Gy per time, following routine radiotherapy of 39 Gy/30f/3weeks. Fourteen cases received total-course conformal radiotherapy with the dose of 59.8 Gy/46f/4.6weeks at 90％ isodose. Results： The reference isodose curves conform to the target in three dimensions in conformal radiotherapy. The acute radiation mucositis, cranial nerve paralysis and trismus incidence were similar in two groups, however, in the hypofractionation group 22％ developed necrosis of nasopharyngeal mucosa, 3.7％ developed reduced optic nerve lesion, whereas none of the hyperfractionation group had these complications. The tumor response rate were both 93％ for two groups. Conclusion： Hyperfraction conformal radiotherapy is effective in the treatment of recurrent nasopharyngeal carcinoma and has relatively lower incidence of radiation complication.

Objective To investigate a reproducible model of severe crush injury (CI) in rats.Methods A total of 50 clean grade male SD rats were randomly (random number) divided into 5 groups.Both hindlimbs of anesthetized rats were compressed by blocks weighing 3.5 kg,for 6 hours and followed by 3 hours of reperfusion on a specially notched device (group SP,n =10),ordinary compression (group NM,n =10) and simple control (group SHAM,n =10).Arterial tension,serum lactate,and potassium (K+),serum myoglobin (MB),aspartate transferase (AST) and alanine transferase (ALT),BUN and Cr were measured at 10 minutes after cannulaton,and 3 hours after release from compression.Muscles and kidneys were evaluated morphologically.Group D and E were treated in the same way and were observed for 72 h to get the survival rate of the NM group and the specially notched compression group.The SPSS 17.0 statistical software was used for statistical analysis,repeated-measures ANOVA analysis for the differences between groups,Kaplan Meier-estimator for survival analysis.Results The Specially notched compression produced a greater increase in serum lactate (F =39.626,P ＜ 0.05),AST (F =24.965,P ＜ 0.05),ALT (F =19.096,P＜0.05),BUN (F=7.938,P＜0.05),CR (F=14.787,P＜0.05) and MB (F=16.840,P ＜0.05) by the end of experiment than NM group and simple control group.The direct cellular damage and ischemia-reperfusion injury were found under microscope.In crush injury caused by specially notched compression there was acute tubular necrosis found at 24 hours after injury.Mortality rate in the NM group was 20％,whereas mortality rate reached 90％ in rats with specially notched compression (P ＜0.05).Conclusions It successfully developed a severe crush injury model in experimental rats,suggesting it is worthwhile to popularization.

ObjectiveTo evaluate the effect of large-spot and low-energy Q switched Nd:YAG laser on melasma,and to observe the changes of melasma lesions with confocal laser scanning microscopy (CLSM) before and after the laser treatment.MethodsTotally,45 patients aged from 24 to 48 years and diagnosed with facial melasma were included in this study,and treated with large-spot and low-energy Q switched Nd:YAG once a week for 10 or more sessions.CLSM was used to estimate the melanin content in melasma lesions before each irradiation and after the last irradiation.ResultsAmong the 45 patients,8 ( 17.78％ ) were nearly cured,25 (55.56％) markedly improved,11 (24.44％) improved,and only 1 (2.22％) unimproved after the laser irradiation.The total response rate was 73.33％.As CLSM showed,there was an increment in melanin granules in melasma lesions compared with the normal skin surrounding melasma lesions,but a reduction in melanin granules was induced by the laser treatment in melasma lesions.ConclusionsLarge-spot and lowenergy Q switched Nd∶YAG laser is substantially effective and highly safe for the treatment of melasma,and CLSM can be used to evaluate the therapeutic effect of laser on melasma.

Objective To detect the gene mutation of thyroid hormone receptor β ( TRβ ) in a family with thyroid hormone resistance syndrome.Methods The genomic DNA was extracted from peripheral blood leukocytes of the patient and his 5 family members.The exons 1-10 ofTRβ gene were amplified by PCR.The products of PCR were sequenced directly to detect the gene mutation.Results Two members of this family were confirmed to have the C y A transition mutation at nucleotide 1642 site within exon 10 of TRβ gene,which was a missense mutation causing the substitution of Proline to Threonine (P453T).The mutation was Heterozygous.Conclusions It was confirmed that the patient has TRβ gene mutation P453T in exon 10.The mutation may lead to the occurrence of thyroid hormone resistance syndrome.

Objective To evaluate the efficacy of multiple injection of morphine combined with phloroglucinol in the treatment of renal colic. Methods One hundred and twenty patients with severe renal colic were equally divided into three groups by random digits table,with 40 cases each group. The group A was administrated with physiological saline 100 ml and phloroglucinol 80 mg intervenous drop infusion combined with morphine 9 mg intravenous injection fractionated into three times, and the group B was administrated with physiological saline 100 ml and phloroglucinol 80 mg intervenous drop infusion,and the group C was administrated with physiological saline 100 ml intervenous drop infusion and morphine 9 mg intravenous injection fractionated into three times. At 10,20 and 40 min after administration, the antalgic efficacy of three groups were analyzed as well as the side effect. Results At 20 min and 40 min both of the total efficacy rates in group A[82.5%(33/40),95.0%(38/40)] were higher than those in group B [62.5%(25/40),80.0%(32/40)] and group C [60.0%(24/40),77.5%(31/40)] (P＜ 0.05),and the colic-exclusion rate in group A was also higher than that in group B and group C (P ＜ 0.05). The difference of the efficacy rates in group A in these two time-points had no statistical significance (P ＞0.05). No increased pain occurred in group A except 1 case of vomiting. Conclusion Multiple injection of morphine combined with phloroglucinol is quick, effective and safe in the early treatment of renal colic.

Objective To investigate the association between polymorphisms of thyroid-stimulating hormone receptor(TSHR)gene intron 1(rs179247, rs12101261)and Graves′ disease(GD)in the China Han population from Xuzhou city, Jiangsu Province. Methods Total 1 066 GD patients and 1 107 control subjects were recruited for genotyping by Taqman probe technique on Fluidigm EP1 platform. Meanwhile, serum concentrations of thyroid hormone and TSH receptor antibodies(TRAb)were determined. Results The rs179247_A, rs12101261_T were significantly associated with GD risk(OR=1.35, 95%CI 1.19-1.54, P=5.92×10-6; OR=1.32, 95%CI 1.16-1.50, P=2.22×10-5). Logistic regression identified that rs179247 was an independent susceptibility locus of GD. Serum TRAb concentration showed a significant difference(P=0.015)among rs179247_AA, AG, and GG genotypes. Conclusion rs179247 and rs12101261 in TSHR intron 1 are both associated with GD, and rs179247 may contribute risk to GD independently. The polymorphism is associated with TRAb, but not with serum concentration of thyroid hormones, age of onset, diffused thyroid goiter, ophthalmic signs, and relapse.

The viral RNA was extracted from purified cymbidium mosa ic virus (CyMV) isolated from Dendrobium orchid cultivated in Hainan island. The gene of the movement protein (MP) was amplified by means of reverse transcripti on-polymerase chain reaction (RT-PCR), and cloned into pGEM-T easy vector. Se quence analysis showed that the gene fragment contained 3 open reading frames (O RFs) which may be encoding 14 kD、12 kD and 10 kD peptides. The nucleotide seque nce of the cloned gene fragment shared 97.8% homology with the MP genes of CyMV isolated from orchids cultivated in Hawaii and Singapore.

Objective To investigate the effect of acute high altitude exposure on lung functions and the relationship between lung functions and acute mountain sickness ( AMS) .Method We collected the lung function and Lewis Lake data of 73 subjects (aged 18 to 26,male) at 400 m above sea-leve and those at high altitude(exposure to 3900 m, 5 d).Results Compared with sea-level, lung functions decreased in forced vital capucity (FVC), maximum midexpiratory flow(MMF), V50, V25 while forced expiratory volume in 1 second(FEV1), peak expiratory flow(PEF), V75 did not change.FVC, FEV1, PEF, MMF were used to analyze the relationship between lung functions and AMS .There was no difference in lung functions between AMS group and NON AMS group at sea-level, but lung functions of AMS group were significantly lower than those of NON AMS group in FVC , MMF at high altitude .There was difference between AMS group and NON AMS group in the rate of change of FVC and MMF .Logistic regression analysis showed that the rate of change of FVC was an independent risk factor , while correlation analysis showed that the change of FVC and the change of oxygen saturation were relevant.Conclusion Lung functions showed restrictive decrease after acute high altitude exposure .Changes of lung func-tions will increase hypoxia and susceptiblity to AMS .

Objective:To investigate the trait of exosomes serected by mesenchymal stem cells derived form orofacial bone(OMMSCs-Exo) and the communication between the exosomes and macrophages.Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.Their cell surface markers were characterized by flow cytometry.the rate of colony formation and the differentiation potential of OMMSCs were evaluated.Exosomes were prepaired from the culture supernatants of OMMSCs(P4-P6).Transmission electron microscopy(TEM) and western blot were used to identify the exosomes.The expression of miRNAs associated with immunity such as miR-223 and miR-let-7c were determined by Real-time RT-PCR.Human peripheral blood mononuclear cells(PBMCs) were isolated from health donor and co-cultured with OMMSCs-Exo.After co-cultured for 24 h,the communication between exosomes and macrophages was tested using a confocal microscope.Results:Human OMMSCs were proved to have the characteristics of mesenchymal stem cells.The diameter of OMMSCs-Exo ranged from 40 to 160 nm.The OMMSCs-Exo expressed CD63 and CD81 and contained miRNAs associated with immune regulation such as miR223 and miR-let-7c.OMMSCs-Exo could be uptaken by macrophages.After co-culture of OMMSCs-Exo with marcrophages for 72 h,miR223 expression in macrophages increased.Conclusion:OMMSCs-Exo has the potential of immune regulation.