In recent years , fungal biofilms related infection have become an increasingly important clinical problem .Many clinically important fungi can form biofilms , including:Candida albicans , Cryptococcus neoformans , Rhodotorula species , Aspergillus fumigatus, Malassezia pachydermatis, Histoplasma capsulatum.Many chronic persistent infections are associated with the fungal bio-film formation.The developmental phases of fungal biofilms are complicated , including adhesion, colonisation, maturation and dispers-al, which are governed by many genes .This review discusses clinical impact of biofilm formation of the common human pathogenic fun -gi, biofilm structure and the molecular mechanisms during the biofilm formation process .

AIM: To study the nutritional role of the normal conditioned medium of astrocytes (NCMA) on the injured cerebral cortex neurons induced by cerebral hypoxia and reoxygenation in vitro. METHODS: The normal and damaged neurons induced by hypoxia and reoxygenation were cultured with normal conditioned medium of astrocytes (NCMA) after the cerebral cortical astrocytes and neurons of rats were isolated. The viability and survival rate of cultured neurons were investigated by MTT method. RESULTS: NCMA increased the viability and survival rate of cultured neurons. CONCLUSION: NCMA has nutritional and supporting roles on cultured neurons.

Objective To study clinical characteristics and diagnostic methods of lymphoma with chest invovement. Methods Twenty-five lymphoma patients with chest involvement were retrospectively analysed, they were all diagnosed in Peking University Third Hospital during 2000 to 2007. The data were collected including clinical manifestations, blood examinations, chest X-ray and CT scan, diagnostic methods and pathologic diagnosis. Results The median age of the 25 patients was 46 years old. Pyrexia(13 cases), weight loss over 10 percent in 6 months(11 cases), cough(10 cases), shortness of breath(9 cases) and painless enlargement of the peripheral lymph nodes(16 cases) were common manifestations. Erythrocyte sedimentation rate and serum lactate dehydrogenase(LDH) level were increased in 72.7% and 81% patients, respectively. The enlargement of mediastinum lymph nodes(16 cases, 64%) was the most common presentation of chest radiography, followed by pulmonary involvement(15 cases, 60%) including infiltration or pulmonary consolidation, mass, multiple nodules, diffuse ground-glass shadow, miliary lesion. There were also presentations of pleural effusion(10 cases, 40%), pericardial effusion(4 cases, 16%), chest wall mass(2 cases, 8%). Eighteen patients(72%) had at least two kinds of these presentations. The appearance of pleural effusion were yellow turbid, bloody or chyliform. Rivaha tests were all positive. The median value of plearal effusion examinations were listed as follows: specific gravity 1.031, total cells 9800×10~6/L, WBC 6.72×10~9/L, lymphocyte 86%, neutrophil 14%, protein 31.4 g/L, LDH 296 U/L,adenosine deaminase (ADA) 67.4 U/L Most patients(16 cases) were diagnosed by surgical biopsy,especialy peripheral lymph nodes biopsy (12 cases). Other patients were diagnosed by ultrasound or CT-guided biopsy (5 cases), video-assisted thoracoscopic pleural biopsy (1 case), video-mediastinoscopic mediastinum lesion biopsy(1 case), bronchial mucosa biopsy through bronchoscope(1 case), bone marrow examination(1 case). All the cases were non-Hodgkin lymphoma except one. Conclusions There was no specific clinical manifestation for lymphoma with chest involvement, but in almost half of patients there were enlargement of not only peripheral but also mediastinum lymph nodes. And there were some characteristics in serum, pleural effusion, chest X-ray and CT scan. Surgical biopsy of peripheral lymph nodes was a simple and convenient method for diagnosis. Micro-invasive biopsy had good diagnostic value for lymphoma with chest involvement, including ultrasound-or CT-guided biopsy for superficial mass, pleura, lung, liver, spleen and deep lymph nodes, video-assisted thoracoseopic and video-mediastinoscopic biopsy for pleura, lung and mediastinum lesions. But bronchial mueosa and lung biopsy during bronchoscopy had a low diagnostic rate for lymphoma.

Objective To construct quality control materials for chlamydia trachomatis (CT) polymerase chain reaction detection and evaluate the stability of the material.Methods The reference regarding the target sequence for CT PCR detection has been reviewed.The ovedap extension technique and molecular cloning techniques were used to construct a recombinant lasmid.Then the recombinant plasmid pTARGETTM-CT was transfected into a HTB-SiHa cells.The cuhured epithelia cells,vere collected as quality control material.Then we evaluated the stability of this material with domestic kits for CT PCR detection.The stability in different conditions were summarized and evaluated.The EQA samples for CT test survey ere prepared from the above prepared cells and distributed to the EQA participants nationwidely.Results Five fragments from CT(178-610),(1219-1993),(2471-3260),(5239-5864),(6722-7499) were cloned into pTARGET TM.The recombinant plasmid was transfected into mammalian cells as a final form for the quality control materials.Real-time PCR analysis howed the original material was positive with domestic chlamydia trachomatis kit(3.21×108 copies/ml).A Series of dilution resulted in the decreased result .The stability testing indicated the quality control materials were stable at least for one month when stored at 4℃,room temperate or 37℃.Conclusions We used several kinds of molecular iology methods such as ovedap PCR and enzyrnatie digest to construct a recombinant plasmid which contained several fragments.The quality control materials for chlamydia trachomatis PCR detection was developed successfully.

Objective Little is known about the role of low oxygen concentration in the formation of the capsule of Cryptococ -cus neoformans .This study was to investigate the effect of hypoxic condition on the capsule size of Cryptococcus neoformans . Methods We cultured 39 Cryptococcus neoformans strains in liquid Sabouraud medium and then incubated them at the normoxic (21%) and hypoxic concentration (1%), respectively.After 72 hours of incubation, we observed the capsules formed at different ox-ygen conditions under the Olympus microscope and the measured their sizes using the Axiovision software . Results Of the total number of Cryptococcus neoformans strains , 30 showed larger capsules in the normoxic than in the hypoxic condition , 17 with statisti-cally significant differences in their diameters (P<0.05), while the other 9 strains exhibited just the opposite results , 4 with statisti-cally significant differences in their diameters ( P<0.05). Conclusion Hypoxia might inhibit the growth of the Cryptococcus neoformans capsule.However, further studies are needed on its correlation with the serum or gene types of the Cryptococcus neofor-mans strain.

Objective Candidas rank as the second factor of deep fungal infection .This study aimed to investigate the rela-tionship between Candidas and melanin by screening clinically isolated melanin -producing Candidas . Methods We incubated 360 strains of clinically isolated Candidas with caffeic acid , DOPA agar , and minimal medium at a temperature of 35℃, with Cryptococcus neoformans as the positive control .We observed the changes in the culture media using transmission electron microscopy (TEM) and electron spin resonance ( ESR) spectroscopy . Results Cryptococcus neoforman produced pigmentation in all the three types of medi -um.At 5 days, a standard strain of Candida albicans SC 5314 turned dark brown in the DOPA agar and minimal medium and , at 10 days, 12 and 15 of the 360 Candida strains turned dark brown in the DOPA agar and minimal medium , respectively, which were not shown to be melanin by TEM and ESR spectroscopy . Conclusion Culture media are not reliable enough for the identification of mel-anin-producing Candidas , and the observations need to be further verified by TEM and ESR spectroscopy .

Eccrine spiradenoma usually occurs as solitary tender nodules. However, it rarely occurs as multifocal localized tumors or has zosteriform distribution. We report the case of a 32-year-old woman with a 20-year history of itchy lesions on the left side of the back and forearm with a zosteriform distribution. Before disease onset, almost no patients have inducing factors, but our patient received an injection beforehand; however, whether this was coincidental or causative remains unknown. The lesions became very itchy after perspiration or eating spicy food, which has never been reported. A literature search revealed 22 cases of multiple segmental eccrine spiradenoma; we summarized the clinical characteristic in order to aid diagnosis and treatment selection.
Adult ; Diagnosis ; Eating ; Female ; Forearm ; Humans

A 48-year-old female presented with a 6-year history of papules and plaques all over the body and with 1-year history of blurred vision in the right eye.Physical examination showed porcelain-white atrophic papules with peripheral erythematous halo and telangiectnsia.She also suffered from exotropia,visual deterioration,visual field defects of the right eye,as well as numbness of the left index finger,thumb and right anterior tibia.Skin biopsies of abdominal lesions revealed dermal necrosis with mucoid degeneration,inflammatory infiltration predominated by lymphocytes around several small blood vessels and occlusion of some blood vessels in deep dermis.Colonoscopy of the whole colon demonstrated scattered patches of hyperemia and erosions with the formation of shallow ulcers.Nerve electromyologram revealed damage to the nerves of right quadriceps femoris muscles.Fecal analysis showed that occult blood was strongly positive.A diagnosis of malignant atrophic papulosis was made based on the characteristic clinical presentation,laboratory and histopathological findings.She was treated with dipyridamole and aspirin for three months,which resulted in no clinical improvement or deterioration.

Objective: To study the influence of conditioned medium of rat brain microvascular endothelial cells on the activity of cortical neurons and the protective effect of Tong Luo Jiu Nao Injection.Methods: We collected the conditioned media from 4 different cultured endothelial cell groups,which were normal endothelial cells,the normal ones and treated with Tong Luo Jiu Nao Injection,the injured ones damaged by simulated cerebral ischemia,as well as the injured ones and treated with Tong Luo Jiu Nao Injection,respectively(N-CM,NT-CM,I-CM,IT-CM).Then,the conditioned medium was added into the cultures of the normal neurons and the damage ones which are injured by simulated cerebral ischemia as well,respectively.The effect of each type of conditioned medium on the activities of neurons was determined through the measurement of MTT and the transduction rate of LDH.Results:(1) N-CM has no obvious effect on the normal neurons,but does show some protective effect on the injured ones by increasing its activity significantly;(2) I-CM could decrease the activity of the normal neurons as well as aggravate the damage on the injured ones,while this injury effect can be reversed remarkably by IT-CM.Conclusion: The paracrine secretion of the brain microvascular endothelial cells might be one of the vital mechanisms in cerebral ischemic injury,indicating that the brain microvascular endothelial cells could be the therapeutic targets of Chinese medicine,which are not able to permeate through the Blood-Brain Barrier.

Objective There have been a few reports on chronically recurrent and generalized superficial mycosis caused by trichophyton rubrum.This article was to investigate the cause, diagnosis and therapy of the mycosis. Methods 5 patients with chron-ically recurrent and generalized superficial mycosis caused by trichophyton rubrum were collected from June 2012 to June 2014 in our hospital.Bacterioscopic examination and cultivation were made on skin lesions of the patients.A typical patient who had 7-year course of desease with toenails seriously infected and widespread skin eruption was selected for histopathology examination on skin lesions, mi-crobiology and molecular biology study on 4 bacterial strains isolated from skin lesions in different parts, and in vitro chemosensitivity assay for drug selection.PCR (rDNA ITS sequence analysis) was performed for diagnosis and early treatment. Results Microscopic examintion on skin lesions demonstrated numerous septate, branched hyphae.Cultivation and molecular biology study identified tricho-phyton rubrum.The strain was identified as trichophyton rubrum by ITS sequence analysis and the isolated strains from different lesions were the same fungal species.Histopathology examination revealed slight hyperplasia of squamous epithelium , epidermal hyperkeratini-zation and the upper dermis presented a sparse perivascular lymphocytic infiltrate.The PAS-stain confirmed the presence of few hyphae in the horny layer.The pathogen of this case was trichophyton rubrum. A combination therapy with systemic itraconazole and topically applied terbinafine hydrochloride cream was successful.A follow-up examina-tion one year later showed no recurrence of symptoms. Conclusion The isolation and identification of pathogen is the key to the diagnosis of chronically recurrent and generalized superficial mycosis, with ad-ditional attention to all or none toenail infection.The therapy should not focus simply on the tinea corporis, while comprehensive treatment combined with chemosensitivity assay is preferred.