Genetic Identification of Spirometra erinaceieuropaei Spargana in Liaoning and Hubei Provinces, PR China.

Spargana were collected from human and frogs in Liaoning and Hubei Provinces, China. PCR amplification and direct sequencing of A cox1 fragment was PCR-amplified from genomic DNA extracted from 7 specimens (5 from humans and 2 from frogs). The cox1 fragment (390 bp) showed 97-100% similarity to the reference sequence of S. erinaceieuropaei and 88-89% to the reference sequence of S. decipiens. There were 1-12 bases different between these worms, but no obvious genetic variation (0-3.3%) to the references. There was little difference of cox1 gene between sparganum samples of humans and frogs (1-3%). This study is the first report on S. erinaceieuropaei spargana from humans in Liaoning and Hubei Provinces.

Genotyping of Pneumocystis jirovecii by Use of a New Simplified Nomenclature System Based on the Internal Transcribed Spacer Regions and 5.8S rRNA Gene of the rRNA Operon.

Genotyping based on internal transcribed spacer 1 (ITS1) and ITS2 of the rRNA operon has played an important role in understanding the transmission and epidemiology of Pneumocystis jirovecii, one of the major opportunistic pathogens in individuals with AIDS and other immunocompromised individuals. The widespread use of this typing system has resulted in several problems, including inconsistent genotype nomenclatures, difficult data transferability, and complicated interpretation of the length variation in multiple homopolymeric tracts. The aim of this study was to establish a new, simplified genotype nomenclature system for P. jirovecii based on the ITS1 and ITS2 sequences. We first analyzed the complete ITS1, 5.8S rRNA gene, and ITS2 sequences (termed ITS1-5.8S-ITS2) in 27 recent P. jirovecii isolates from China and identified 18 unique genotypes. Subsequently, we performed a comprehensive classification of more than 400 ITS1- and ITS2-related sequences from GenBank and an in-depth evaluation of the length variation of multiple homopolymeric tracts within ITS1-5.8S-ITS2. Integration of the results from these analyses led to a new, simplified genotype nomenclature system including 62 unique ITS1-5.8S-ITS2 genotypes, simply designated types 1 through 62. This new system offers several advantages over traditional ITS1- and ITS2-based typing systems, including a simpler analysis and interpretation process, a higher discriminative power, and no limitation in assigning potential new genotypes. This new system is expected to facilitate the standardization of P. jirovecii genotyping and easy data exchanges across different laboratories.

Clinical experience of individual surgical therapy in hypertensive basal ganglia hemorrhage.

BACKGROUND: The aim of this article is to summarize the clinical experience stemming from the administration of different surgical therapies in hypertensive basal ganglia hemorrhage (HBGH) patients. METHODS: A series of 87 patients with HBGH who had received surgical therapy individually were enrolled in this study. The surgical therapies were stereotactic aspiration (SA), stereotactic aspiration plus fibrinolytic therapy (SA+F) and microsurgery with small bone window (MS), respectively. The outcomes of the patients were evaluated by evolution of hematoma evacuation, activities of daily living (ADL) scale, mortality and complications. RESULTS: We found that there was no significant difference in the 24-hour evacuation rate, mortality and complication rate among treated groups (P>0.05). Though patients in level III and level IV of ADL scores were significantly different among the three groups, the overall ADL scale result demonstrated a similar ADL result. CONCLUSIONS: HBGH patients should be treated with an individualized surgical approach based on their condition and on the CT morphology of the hematoma.

Synthetic p55 tandem DNA vaccine against Pneumocystis carinii in rats.

Pneumocystis spp. are opportunistic fungal pathogens that are closely associated with severe pneumonia and pulmonary complications in patients with impaired immunity. In this study, the antigenic epitopes of the gene encoding the 55 kDa antigen fragment of Pneumocystis (p55), which may play an important role in Pneumocystis pneumonia, were analyzed. A gene containing tandem variants of the p55 antigen was synthesized and named the tandem antigen gene (TAG). TAG's potential as a DNA vaccine was assessed in immunosuppressed rats. Immunization with p55-TAG DNA vaccine significantly reduced both the pathogen burden and lung-weight to body-weight ratios. Additionally, p55-TAG vaccination in immunosuppressed rats elicited both cell-mediated and humoral immunity.

Investigation of Pneumocystis jirovecii colonization in patients with chronic pulmonary diseases in the People's Republic of China.

BACKGROUND: The detection of Pneumocystis jirovecii DNA in respiratory specimen from individuals who do not have signs or symptoms of pneumonia has been defined as colonization. The role of P. jirovecii colonization in the development or progression of various lung diseases has been reported, but little information about P. jirovecii colonization in patients is available in the People's Republic of China. OBJECTIVE: To determine the prevalence of P. jirovecii colonization in patients with various pulmonary diseases, including the acute and stable stage of COPD, interstitial lung diseases, cystic fibrosis, and chronic bronchiectasis. MATERIALS AND METHODS: A loop-mediated isothermal amplification (LAMP) and a conventional polymerase chain reaction (PCR) method for detecting P. jirovecii were developed. Ninety-eight HIV-negative patients who were followed-up and who had undergone bronchoscopy for diagnosis of various underlying respiratory diseases were included in the study. Sputa of these patients were analyzed with LAMP amplification of P. jirovecii gene. In addition, conventional PCR, Giemsa and Gomori's methenamine silver nitrate staining assays were applied to all specimens. RESULTS: The sensitivity and specificity test showed that there was no cross-reaction with other fungi or bacteria in detecting the specific gene of P. jirovecii by LAMP, and the minimum detection limits by LAMP was 50 copies/mL. P. jirovecii DNA was detected in 62 of 98 (63.3%) sputa specimens by LAMP assay and 22.45% (22/98) by conventional PCR. However, no P. jirovecii cysts were found by Giemsa and Gomori's methenamine silver nitrate in all of gene-positive specimens. CONCLUSION: The results of our study showed that prevalence of P. jirovecii colonization is particularly high in patients with chronic pulmonary diseases in the People's Republic of China, and the LAMP method is better for evaluation of the colonization of P. jirovecii in sputum specimen than conventional PCR.

Mini-Review of Published Reports on Coccidioidomycosis in China.

Coccidioidomycosis is a deep mycotic infection endemic to Americas. Although it has also been reported to occur in non-endemic areas, it has rarely been reported in China. We reviewed the literature on case reports of coccidioidomycosis in China from 1958 to 2015. A total of 30 cases were reported from 11 provinces in China, and 23 (76.7%) cases were men, and 23 (76.7%) were in 30-60 years old. Twenty-seven (90.0%) cases were reported from southern China. Twenty-four (80.0%) cases had no history of exposure in endemic areas. Three cases were immunosuppressed, four cases had chronic disease, and 23 cases (76.7 %) were otherwise healthy. Twenty cases (66.6%) had related lung infection, six had skin infection, three cases were in lymph node, and one in the cornea, one in the bone and joint, and three had systemic disseminated infection. All of the 30 cases were diagnosis upon finding spherules in histological examination. Interestingly, 12 (40.0%) patients underwent surgery and removed the focus of infection because they were misdiagnosed or suspected of tumor or cancer previously. Fifteen cases (50.0%) were treated with amphotericin B, fluconazole, clotrimazole or ketoconazole. Diagnosis of coccidioidomycosis is complex, and misdiagnosis can occur easily in non-epidemic areas such as China.

A pediatric case report and literature review of thelaziosis in China.

Thelaziosis is a zoonosis with regional epidemics, mainly present in oriental countries, with a majority of cases reported from China. We report a pediatric case of thelaziosis and present a literature review of Chinese case reports. The epidemiological features and clinical characteristics of thelaziosis in China are presented.

Emergency microsurgery for aneurysmal subarachnoid hemorrhage coexisting with brain hernia.

AIM: This work aims to summarize the experience in emergency microsurgery for patients with aneurysmal subarachnoid hemorrhage (SAH) that coexisted with brain hernia. MATERIAL AND METHODS: A total of 26 patients with aneurysmal SAH complicated by brain hernia were subjected to emergency aneurysm clamping + decompressive craniectomy. 10 patients were diagnosed with aneurysmal SAH coexisted with brain hernia before operation. RESULTS: Four patients had a carotid aneurysm, 15 had a middle artery aneurysm, and 7 had an anterior communicating aneurysm. The surgical procedure was performed smoothly for all patients. The patients were followed up for 3 months to 6 months. According to Glasgow outcome scaling, five patients recovered well, six suffered from moderate disability, five were subjected to severe disability, five were in plant survival, and 5 died. CONCLUSION: Emergency surgical exploration is an effective treatment method for patients.

Two cases of human thelaziasis as confirmed by mitochondrial cox1 sequencing in China.

Two special cases of human thelaziasis were reported in China: an old farmer with heavy infection by 36 worms and a 7-year-old boy with infection by eight worms. Thelazia callipaeda was morphologically identified and confirmed by mitochondrial cox1 gene sequencing.

Trichuriasis diagnosed by colonoscopy: case report and review of the literature spanning 22 years in mainland China.

Trichuriasis is soil-source parasitic disease, usually endemic in warm, humid, tropical and subtropical countries. We report a case of trichuriasis diagnosed by colonoscopy performed in a non-endemic area and review the literature on the colonoscopic diagnosis of Trichuris trichiura spanning 22 years (1989-2011) in mainland China. A total of 4382 trichuriasis cases were diagnosed by colonoscopy out of 41,337 patients who underwent colonoscopy. Most of the patients were cured by unloading the whipworms using colonoscopy pincers. We strongly suggest colonoscopy as a useful diagnostic and treatment tool, especially when the patient is lightly infected or is infected by only a few male worms with no eggs in the stool. The geographical distribution, parasite burden, and the parasitizing locations of T. trichiura in the intestine of the human body were analyzed, and the clinical signs and symptoms, as well as the utility of colonoscopy in the diagnosis and treatment of trichuriasis are discussed.

[Allele genotype analysis of Plasmodium vivax merozoite surface protein 1 (PvMSP-1) from Dangdong, Liaoning Province].

Nested PCR method was used to amplify the Plasmodium vivax merozoite surface protein 1 (PvMSP-1) gene fragment containing the ICB5 and ICB6 region from Plasmodium vivax in Liaoning Province. The PCR products were digested by Pvu II restriction endonuclease and the digested fragments were observed by 1.5% agarose gel electrophoresis, and all followed by sequencing analysis and comparison. In 11 field isolates of P. vivax, two kinds of DNA fragments with 470 and 400 bp were produced respectively. After PvuII digestion, two Sal-1 type fragments (120 and 350 bp) were obtained from 5 samples of 470 bp. Single band of 400 bp appeared in 1 samples as Belem type. Two bands of 120 and 280 bp appeared from another 1 sample as recombination type III, and other 4 bands with 120 and 240 bp as Korean isolate. The principal types of PvMSP-1 alleles exist in malaria endemic areas in Liaoning Province with no mixed infection of two different type alleles.

Retrospective study of Pneumocystis pneumonia over half a century in mainland China.

A retrospective study was performed on case reports of Pneumocystis pneumonia (PCP) from 1959 to 2009 in mainland China. The epidemiological characteristics of PCP over half a century were investigated over two time spans. The first was from 1959, when the first incidence of PCP was reported, to 1984, before the emergence of AIDS in mainland China. The second was from 1985, when the first AIDS case was reported in mainland China, to the end of 2009. A total of 2351 PCP cases were reported during these two time spans, covering a 51-year period. Only seven PCP cases were reported during the first time span. Six were diagnosed by autopsy, accordingly without treatment, whilst the other was diagnosed by open lung biopsy in a living patient who eventually recovered following treatment with sulfadiazine and pyrimethamine. The other 2344 PCP cases were reported during the second time span (1985-2009) from 21 provinces, four municipalities and three autonomous regions. Among the 2344 PCP cases, 70.22 % (1646/2344) were identified together with human immunodeficiency virus (HIV) infection or were in AIDS patients. The remaining 698 non-HIV-infected patients had undergone organ transplantation, had other underlying diseases such as malignancy or hypoimmunity, or had undetermined diagnosis. The results of statistical analysis indicated that AIDS was the most common underlying disease of PCP for patients <1 year and >14 years. For patients aged between 1 and 14 years, haematological malignancy was the most common underlying disease. The trend of the underlying diseases changed with time, showing that the number of PCP patients afflicted by HIV/AIDS increased dramatically, reaching almost threefold during the most recent 5 years compared with the level of the previous 10 years. The number of patients undergoing organ transplantation or with other underlying diseases rose constantly, but the number of malignancies tended to decline from 1995-2004 to 2005-2009. During the second time span (1995-2009), most of the patients (97.61 %) were diagnosed alive and only 56 cases (2.39 %) were identified by autopsy. The mortality of PCP patients treated with anti-Pneumocystis drugs was 14.61 % for those with HIV/AIDS and 15.84 % for those without HIV/AIDS. For the PCP patients without anti-Pneumocystis treatment, all (100 %) of the HIV/AIDS-associated PCP patients died, whilst 13.79 % (4/29) of non-HIV-infected PCP patients survived. These data from epidemiological investigation of PCP in China over a period of half a century may provide useful information for prevention and the development of treatment of PCP.

[Progress on molecular biology of Pneumocystis--for hundred years since its discovery].

2009 marks the 100th anniversary of the first description of Pneumocystis, an organism that was initially classified as a protozoan and later in 1988, was phylogenetically reclassified in the fungal kingdom. The organism was ignored for its first 50 years but that has subsequently been recognized as an important pathogen in immunocompromised patients, especially patients with acquired immune deficiency syndrome(AIDS). This paper reviewed recent progress on the molecular biology, involving in the life cycle and metabolism of the organism.

[In vitro observation on effect of nitric oxide on exflagellation of Plasmodium yoelii].

OBJECTIVE: To observe the effect of nitric oxide (NO) on exflagellation of malaria parasite. METHODS: The level of parasitemia and gametocytaemia in DBA/2 mice infected with Plasmodium yoelii 17XL was measured by scanning Giemsa-stained blood smears, and the NO level in culture supernatant of splenocytes was checked using Griess reaction. The mice were injected with different doses of NO donor (NOC5) on day 4 post-infection, and control mice were injected with NOC5 precursor. On day 6 post-infection, mice were injected with NOS inhibitor (L-NMMA), and control mice were injected with D-NMMA and PBS, respectively. Blood samples were collected from tail vein of mice before injection, 30 and 60 min after being injected with NOC5 and NOC5 precursor, 4 and 8 h after being injected with L-NMMA, D-NMMA, and PBS respectively. Exflagellation number of gametocytes in blood culture was counted under microscope. Results The NO level in culture supernatant of splenocytes from mice on day 4 and 6 post-infection was 16.5 mmol/L and 30.4 mmol/L, and exflagellation number was 11.33 and 0.66, respectively. The number of exflagellation in parasitized erythrocytes, obtained from mice on day 4 post-infection, was 5.33 and 2.66, respectively, 30 and 60 min after injection of 1 mg NO donor (NOC5), significantly lower than that of the control (P<0.01). The number of exflagellation in parasitized erythrocytes derived from mice on day 6 post-infection was 1.83, 8 h after the injection of NOS inhibitor (L-NMMA), which was significantly higher than that of the control (P<0.01). CONCLUSION: NO is a major effector molecule resulting in natural transmission-blocking of malaria parasite by directly inhibiting exflagellation of male gametocytes.

[Measurement of morphological parameters of Cryptosporidium muris oocysts by digital image processing technology].

OBJECTIVE: To establish and analyze the morphological parameters of the oocysts of Cryptosporidium muris for defining their morphological change. METHODS: Oocysts were collected from KM mice (immunodepressed by dexamethasone for 10 days) and examined with modified acid-fast staining. Images of 1 190 oocysts were acquired by photograph system. The length, width, perimeter, area and equivalent diameter of the oocysts were obtained by computer digital image processing system and analyzed by SPSS software (Version 11.0). RESULT: The average length of the oocysts was 5.93 microm, ranging from 3.36 microm to 8.51 microm in 95% confidence interval of them. The average width was 4.96 microm, ranging from 3.26 microm to 6.66 microm in 95% confidence interval of them. The average perimeter was 18.03 microm and the average area was 16.08 microm2. CONCLUSION: Data obtained from the computer system are objective and precise, offering scientific foundation for measuring the oocysts and for identifying Cryptosporidium spp.

Exposure of immunocompetent adult mice to Pneumocystis carinii f. sp. muris by cohousing: growth of P. carinii f. sp. muris and host immune response.

There has been emerging evidence that immunocompetent hosts can harbor Pneumocystis in their lungs. The purpose of this study was to determine the kinetics of Pneumocystis carinii f. sp. muris infection in adult immunocompetent mice and the host immune response to the organisms. To accomplish this, we exposed adult immunocompetent mice to SCID mice infected with P. carinii f. sp. muris by cohousing. We found that P. carinii f. sp. muris was detectable in the lungs of cohoused immunocompetent mice by PCR by 3 weeks after the beginning of cohousing. At about 4 weeks of cohousing, P. carinii f. sp. muris was readily detectable in the lungs of mice by microscopic techniques. Also at this time, P. carinii f. sp. muris-specific immunoglobulin G was found in the sera of the mice, and CD62(low) CD4- and CD8-positve T cells accumulated in the lungs. Shortly after this immune response, the P. carinii f. sp. muris organisms were cleared from the lungs. Adult mice cohoused for only 1 week also contained P. carinii f. sp. muris cysts detectable by silver staining at 5 and 6 weeks after the beginning of cohousing. We also found that the P. carinii f. sp. muris organisms grew to greater numbers in the lungs of BALB/c mice than in those of C57BL6 mice. This indicates that immunocompetent hosts develop a mild infection with P. carinii f. sp. muris which resolves in 5 to 6 weeks when there is a detectable immune response to the organism. Once an acquired immune response was initiated, the P. carinii f. sp. muris organisms were quickly eliminated without clinical signs of disease.

[Susceptibility and immune responses to Pneumocystis carinii infection in different strains of mice by contagious transmission].

OBJECTIVE: To explore the susceptibility of different strains of mice to P. carinii infection and the host immune response to the organism. METHODS: C57BL/6 and BALB/c strains of mice (15 each) were exposed to SCID mice infected with P. carinii by co-housing. Observations were made on the number of parasites in the lungs, level of CD4+ T cell and CD8+ T cell in BALF, and serum IgG at 4, 5 and 6 wk after contagion. RESULTS: The number of P. carinii grew in the lungs of BALB/c mice was found much greater than those in C57BL/6 mice. A few number of P. carinii cysts were detected in the lungs of both strains of mice by 4 wk after co-housing, the number of cysts increased at 5 wk in the lungs of BALB/c mice but not in that of C57BL/6 mice. P. carinii-specific IgG in the sera and high level of CD62low CD4- and CD8-positive T cells in the lungs were found at 5 wk. The parasites were cleared from the lungs at 6 wk in all infected mice, shortly after acquired immune response was initiated. CONCLUSION: BALB/c mice are more susceptible to P. carinii than the C57BL/6 mice by natural transmission, and all the immunocompetent mice can clear the infection of P. carinii by cellular and humoral immune responses.