| Objective: Cryptococcal meningitis (CM) is a central nervous systeminfection disease caused by Cryptococcus neoformans (Cn), which violatedthe meninges and/or brain parenchyma. CM is mainly seen inimmunocompromised patients and a small number of apparentlyimmunocompetent hosts. Patients in the foreign countries with cryptococcalmeningitis mostly are HIV-positive patients. While, at home patients withcryptococcal meningitis mostly are patients who was sick. These primarydiseases include organ transplantation, immunosuppressive agents andhormones were using long term. In recent years, a number of factors havecontributed to this increase, such as the using of hormonal,immunosuppressive agents and broad-spectrum antibiotics, organtransplantation, intracranial the invasive examination, AIDS epidemic as wellas raise the level of laboratory testing. Currently, Cryptococcal meningitismortality remains high, due to the lack of antifungal agents and the occurrenceof intracranial complications. Studies have shown that the outcome ofcryptococcosis depends on the immune status of the infected individual andthe cytokine pattern generated in response to the pathogen. In this study weexamined theβ-arrestin2levels in the cerebrospinal fluid of CM patients tofurther evaluate their value in the Pathogenesis of Cryptococcal meningitis,and Provide new therapeutic targets for the treatment of cryptococcalmeningitis. At the same time we researched the endoplasmic reticulum usingthe laser scanning confocal microscope (LSCM).Methods：All patients were from either department of neurology andpediatric inpatients or outpatients in the Second Hospital of Hebei MedicalUniversity during2009June-2012June. These50patients were divided intotwo groups. One group is case group (25cases): Cryptococcal meningitis; and the other group is control group (25cases):22patients had vascular headache,1of these patients had ischemic cerebrovascular disease,1patients hadHysteria,1patients had bronchopneumonia. The case group included twogroups: pre-treatment group and after treatment group.All the cases were conducted lumbar puncture. The specimens weresubmitted to test cerebrospinal fluid routine, biochemical, cytology (MGG dye)and aricine blue staining. The levels of β-arrestin2in cerebrospinal fluid weredetermined under the ELISA theorem. In the immunofluorescence staining,FITC and Hoechst were used to individually tagged anti ATF4polyclonalantibodys and the cell nucleus DNA. Than observed the cells under the laserscanning confocal microscope (LSCM).Results:1Clinical dataCryptococcal meningitis patients might manifest as headache, fever,meningeal irritation sign, raised intracranial tension, consciousness disordersand seizures etc. All of the25CM patients had a headache (100%). Theclinical data of2patients was missing,18of the others had meningealirritation sign(78.26%),17patients had lesions of cranial nerves includingoptic never, acoustic never and abducent never(21.74%), consciousnessdisorders was occurred in6patients(26.09%), seizures were observed in2patients(8.70%).Underlying disease: There were17cases of24CM patients withunderlying disease(68%).The situation of the count of Cryptococcus neoformans in thecerebrospinal fluid: the Cryptococcus neoformans were disappeared in thecerebrospinal fluid in4patients (16%). Cryptococcus neoformans in15patients were significantly reduced (60%). While the count of Cryptococcusneoformans in6patients were not reduced (24%).2patients of25weredie(8%).2Cerebrospinal fluid routine, biochemical examination resultsBecause of the lack of the clinical data of two patients, we did a survey of the Cerebrospinal fluid routine, biochemical examination results of23cases ofpatients with cryptococcal meningitis. The pre-treatment group ofcryptococcal meningitis patients: the average number of the total cells was138.52±160.08×106/L;the average number of white blood cells was53.22±59.07×106/L;the average glucose was2.55±1.06mmol/L; the averageprotein was0.68±0.47g/L; the average chloride was118.95±6.15mmol/L. Thetreatment group of cryptococcal meningitis patients: the average number ofthe total cells was201.43±525.02×106/L;the average number of white bloodcells was16.04±28.47×106/L;the average glucose was3.28±1.20mmol/L; theaverage protein was0.54±0.24g/L; the average chloride was124.43±5.56mmol/L. In the control group, the average numbers respectivelywere total cells9.43±17.04×106/L; white blood cells0.91±1.31×106/L;glucose3.28±0.43mmol/L; protein0.29±0.08g/L; chloride123.63±2.64mmol/L.The total cells count, white blood cells count and protein content in thepre-treatment group of cryptococcal meningitis were obviously higher than thecontrol group (P﹤0.05), there were statistical significances; The content ofglucose and chloride in the pre-treatment group of cryptococcal meningitiswere lower compared with the control group (P﹤0.05),and the differenceswere statistically significant. But no significant differences were detectedbetween the pre-treatment group and treatment group of cryptococcalmeningitis (P＞0.05).3The characteristic of cerebrospinal fluid cytologyThe treatment group compared with the pre-treatment group withincryptococcal meningitis patients, the Cryptococcus neoformans ofCerebrospinal fluid of4cases was disappeared; pathogen of15cases wassignificantly reduced; and6cases were not significantly improvement. In thecase group, the data of cerebrospinal fluid cytology of3patients was missing.Of the22cases of CM patients, before treatment the count of neutrophi ofcerebrospinal fluid was0-90%, the median was11.5%; after treatment thecount of neutrophi was significantly decrease:0-8.7%, the median was1%. And significant differences were detected between the pre-treatment groupand treatment group of CM（P=0.002＜0.005）.4The content determination of β-arrestin2in cerebrospinal fluidThe pre-treatment group of cryptococcal meningitis patients: the range ofβ-arrestin2levels in cerebrospinal fluid was (16.59～137.55)pg/ml, averagecontent35.45±25.41pg/ml; The treatment group of cryptococcal meningitispatients: the range of β-arrestin2levels in cerebrospinal fluid was (13.39～35.36)pg/ml, average content16.57±6.05pg/ml; In the control group: therange of β-arrestin2levels in cerebrospinal fluid was (12.26～19.04)pg/ml,average content15.39±2.02pg/ml; By comparing the pre-treatment group ofcryptococcal meningitis with the treament group and the control group,differences were statistically significant. But no significant difference wasdetected between the treatment group and the control group.In the CM group, the counts of Cryptococcus neoformans of19patientswere significantly reduced: Before treatment the level of β-arrestin2incerebrospinal fluid was （16.59～137.55） pg/ml, average content was35.89±28.55pg/ml; After treatment the level of β-arrestin2in cerebrospinalfluid was（13.39～33.11）pg/ml, average content was18.17±5.57pg/ml;While6patients were not significantly improvement: Before treatment thelevel of β-arrestin2in cerebrospinal fluid was（19.50～41.41）pg/ml, averagecontent was25.72±8.46pg/ml; After treatment the level of β-arrestin2incerebrospinal fluid was （14.97～35.36） pg/ml, average content was20.77±7.61pg/ml. Before treatment, no significant differences were detectedwithin these two groups. Also after treatment, no significant differences weredetected within these two groups.5The changes of ATF4in the endoplasmic reticulum of the CSF cellImmunofluorescence staining:4CM patients and3control cases wereobserved: In the CM group, ATF4located in the nucleus, and the content ofATF4was significantly increased; In the control group, ATF4located in thecytoplasm, and the content of ATF4was extremely low.. Conclusions:1The clinical manifestation of cryptococcal meningitis is diverse, but allof these are nonspecific. And the mortality rate remains unacceptably high.2Cryptococcus neoformans is opportunistic pathogen that causesopportunistic life-threatening meningoencephalitis mainly inimmunocompromised patients, so immunocompromised individuals’ clinicalmanifestations are meningitis should be highly vigilant against cryptococcalmeningitis.3In patients with cryptococcal meningitis, the β-arrestin2levels in thecerebrospinal fluid of pre-treatment group was higher than the treatment group;and was also higher than the control group. There were significant differencesin statistics.4By comparing the improvement group with the no-improvement group,no significant differences were detected. More cases is needed in order tostudy.5In the early times, the count of neutrophi in the cerebrospinal fluidwhich was collected from cryptococcal meningitis patients was significantlyhigher than after treatment. The difference was statistically significant.6In the CM patients’ CSF cells, the changes of the endoplasmicreticulum stress factor ATF4was found. It is indicating that endoplasmicreticulum stress participate in the pathogenesis of CM. |
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