Cryptococcus neoformans are most common Fungi in central nervous system infection.Incidence of cryptococcal meningitis is on the rise in recent years.The use of antifungal drugs must control majority of cryptoeoccosis,and at least one-third of patients have experienced failure of antifungal therapy,and mortality and disable rate are on the rise.Long-term heavy use of antifungal drugs cause cryptococcus resistance phenomenon,and it is the main cause of treatment failure.Therefore,the study of cryptococcus resistance mechanism is imminent.This review summarizes the research progresses of cryptococcus resistance mechanism in recent years,aiming to provide theoretical guidance for the research of new antifungal drugs and providing basis for individual treatment.

OBJECTVE:To investigate the clinical effect and safety of pregabalin combined with gabapentin in the treatment of central pain after cerebral infarction.METHODS:One hundred and fifty patients with central pain after cerebral infarction in our hospital from Jan.2010 to Dec.2015 in our department were randomly divided into group A,B,C,with 50 cases in each group.Group A was given Pregabalin capsule 75 mg,bid combined with Gabapentin capsule 0.1 g,tid;group B was given Pregabalin capsule 75 mg,bid;group C was given Gabapentin capsule 0.1 g,tid;3 groups were treated for 4 weeks.VAS score,NRS score,PSQI and SF-36 score were observed among 3 groups before and after treatment to evaluate clinical efficacies of 3 groups;the occurrence of ADR were recorded in 3 groups.RESULTS:The clinical total response rate of group A,B,C were separately 94.00％,74.00％,70.00％.The clinical total response rate of group A was significantly better than that of group B and C,with statistical significance (P＜0.05).After treatment,VAS score of group A,B,C were separately(3.87 ± 0.74),(5.10 ± 1.26),(5.03 ± 1.23);NRS score were separately (3.91 ± 0.88),(5.29 ± 1.25),(5.37 ± 1.30);VAS score and NRS score of group A were signifi cantly lower than group B,C and before treatment,with statistical significance (P＜0.05);PSQI score of group A,B,C were separately(4.03 ± 0.85),(5.92 ± 1.16),(5.83 ± 1.11);SF-36 score were separately (372.84 ± 73.25),(348.07 ± 60.54),(345.67 ± 59.72);PSQI score and SF-36 score of group A were significantly better than group B,C and before treatment,with statistical sig nificance (P＜0.05).There was no statistical significance in the incidence of ADR among 3 groups (P＞0.05).CONCLUSIONS:Compared with pregabalin and gabapentin alone,pregabalin combined with gabapentin in the treatment of central pain after cerebral infarction can efficiently relieve the perceived pain,improve sleep quality and daily life quality and not increase the risk of ADR;therefore,drug combination plan is recommended for patient with central pain after cerebral infarction,especially with poor effect of two single drug.

Objective To purify native and recombinant heparin-binding hemagglutinin(HBHA)protein,and investigate the activity of HBHA polyclonal antibody against aggregation of Bacillus CalmetteGuerin(BCG)induced by HBHA.Methods After growing BCG to the stationary phase in the 7H9 liquid medium,the native HBHA protein(nHBHA)was obtained by CL-6B column chromatography.At the same time,the HBHA gene fragment was cloned and expressed by transforming Escherichia coli BL-21.Then the polyclonal antibody against rHBHA was prepared by immunizing rabbit.Different comcentration of the HBHA protein was added to the BCG liquid medium,and the aggregation of the BCG was observed.Then,add the HBHA protein that incubated with anti-HBHA antibodies to the BCG culture medium and observe the aggregation of BCG.Results The purity of native HBHA was 99% and the concentration was 1.016 mg/ml.The expressed product contained 36% of total somtic protein.After purified,the purity of the recombinant HBHA protein was 97.1% and the concentration was 10.98 mg/ml.Both the rHBHA and nHBHA could induce the aggregation of BCG.When then concentration of nHBHA is 0.2μg/ml,BCG could be induced to aggregate,while the rHBHA concentration is 2μg/ml could induce the aggregation.Both aggregations could be suppressed by the polyclonal antibody against rHBHA.Conclusions The native and recombinant HBHA are successfully obtained.It is proved that the rHBHA could induce the aggregation of BCG similar as nHBHA,and polyclonal antibody against rHBHA could also suppress the activity of nHBHA.It suggested that rHBHA could be further used in clinical diagnosis and vaccination.

Objective To evaluate the application effect of nursing management in the follow-up of patients with recurrent acute pancreatitis (RAP).Methods During January 2016 and January 2017,the clinical data of RAP patients admitted in Changhai Hospital of Navy Medical University was collected.Selfcontrol method was used to establish a prospective cohort study.Meticulous nursing management mode was applied to follow up the patients for 12 months.Patients' compliance,symptom improvement,patients' satisfaction and other markers are evaluated using questionnaires to assess the effect of meticulous nursing management once every 3 months.The data over 12 months were collected.Results Sixty-four RAP patients were finally included.The average follow-up period was 19.7 ±3.4 months.Before admission,the average disease course was(4.21 ± 3.95) years.30 patients (46.9％)had a history of drinking;33 patients (51.6％)had a history of smoking.25 patients (39.1％) had abnormal blood lipid and 40 patients (62.5％) had the dietary habit of high lipid and high protein.The etiology included alcohol (n =10,15.6％),high lipid (n =7,10.9％),biliary diseases (n =1,1.6％),idiopathic causes (n =46,71.9％) and abnormal BMI (n =49,76.6％).After the meticulous nursing management,the frequency of pancreatitis,VAS score of pain and alcohol intake situation was obviously lower than before intervention [(0.50 ±0.85) times/year vs(2.77 ±2.52)times/year;(1.84 ±2.54)vs(6.47 ±2.15);2 cases vs 30 cases].The number of patients who took medicine on time and BMI were obviously decreased[(52 cases vs 13 cases;(23.26 ± 3.85) kg/m2 vs (21.92 ± 4.27)kg/m2)],and the differences were statistically significant (P ＜0.01).Patients' satisfaction with this nursing model was 4.90 ± 0.56.Conclusions The implementation of the new meticulous nursing management model can effectively alleviate patient's symptom condition and improve the compliance behavior and life quality of the patients.

Objective:To explore whether the lipopolysaccharide (LPS)-induced modification of O-linked N-acetylglucosamine (O-GlcNAc) is involved in the inflammatory signaling pathway of endothelial cells.Methods:Human umbilical vein endothelial cells (HUVEC) were cultured in vitro, and cells in logarithmic growth phase were used for experiments. Cells were divided into blank control group, LPS group (2 000 mg/L LPS), O-GlcNAc transferase (OGT) overexpression (OGT-OE)+LPS group (plasmid transfection OGT+2 000 mg/L LPS), protein kinase C (PKC) inhibitor+LPS group (10 μmol/L Go 6983+2 000 mg/L LPS), RhoA inhibitor+LPS group (40 μmol/L Rhoin hydrochloride+2 000 mg/L LPS), phosphatidylinositol-3-kinase (PI3K) inhibitor+LPS group (1 μmol/L SL-2052+2 000 mg/L LPS), serine/threonine kinase (Akt) inhibitor+LPS group (10 μmol/L PP2+2 000 mg/L LPS) and small interfering RNA (siRNA) treated Akt (si-AKT)+LPS group (si-Akt+2 000 mg/L LPS). After 24 hours of LPS treatment, real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect the transcription levels of inflammatory cytokines [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1)]. The protein expression or phosphorylation of OGT, O-GlcNAc, Akt, extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein kinase (p38MAPK), nuclear factor-κB p65 (NF-κB p65), and signal transducer and activator of transcription 3 (STAT3) were determined by Western blotting. Results:Compared with the blank control group, the expression of OGT and the modification of O-GlcNAc in the LPS group were decreased, while the expressions of phosphorylated ERK, p38MAPK, and STAT3 were increased, and the transcript levels of inflammatory cytokines were also significantly increased [IL-6 mRNA (2 -ΔΔCt): 4.71±0.60 vs. 1.03±0.29, TNF-α mRNA (2 -ΔΔCt): 1.89±0.11 vs. 1.04±0.35, ICAM-1 mRNA (2 -ΔΔCt): 2.06±0.18 vs. 1.02±0.21, VCAM-1 mRNA (2 -ΔΔCt): 2.94±0.57 vs. 1.01±0.17, all P < 0.05], indicating that LPS could decrease O-GlcNAc modification, activate inflammatory signaling pathways and increase inflammatory cytokines expression. Compared with the LPS group, the expressions of phosphorylated ERK, p38MAPK, NF-κB p65, and STAT3 in the endothelial cells of the OGT-OE+LPS group were decreased, and the expression of inflammatory factors were significantly decreased [IL-6 mRNA (2 -ΔΔCt): 0.12±0.01 vs. 0.90±0.17, TNF-α mRNA (2 -ΔΔCt): 0.31±0.01 vs. 0.91±0.14, ICAM-1 mRNA (2 -ΔΔCt): 0.64±0.02 vs. 1.13±0.16, VCAM-1 mRNA (2 -ΔΔCt): 0.11±0.01 vs. 0.93±0.11, all P < 0.05], indicating that the increase of OGT level could inhibit the partial activation of the endothelial inflammatory signal pathway under the LPS stimulation. Compared with the blank control group, the phosphorylation level of Akt in the LPS group was increased. Compared with the LPS group, both OGT expression and O-GlcNAc modification were down-regulated after pretreatment of PKC inhibitor, RhoA inhibitor, PI3K inhibitor, or Akt inhibitor. Compared with the LPS group, the transcript levels of IL-6, TNF-α and ICAM-1 in the PP2+LPS group were significantly decreased [IL-6 mRNA (2 -ΔΔCt): 1.46±0.16 vs. 3.55±0.87, TNF-α mRNA (2 -ΔΔCt): 0.98±0.14 vs. 1.76±0.10, ICAM-1 mRNA (2 -ΔΔCt): 1.39±0.24 vs. 2.04±0.13, all P < 0.05], but there was no significant change in VCAM-1. Compared with the LPS group, the expression of OGT and O-GlcNAc modification in the si-Akt+LPS group were decreased, while the transcript levels of inflammatory cytokines were also significantly decreased [IL-6 mRNA (2 -ΔΔCt): 0.75±0.03 vs. 0.99±0.09, TNF-α mRNA (2 -ΔΔCt): 0.69±0.01 vs. 1.10±0.08, ICAM-1 mRNA (2 -ΔΔCt): 0.76±0.01 vs. 0.99±0.02, VCAM-1 mRNA (2 -ΔΔCt): 0.93±0.08 vs. 1.20±0.21, all P < 0.05], indicating that Akt participated in the action process of LPS on OGT and affected the inflammatory factor expression. Conclusions:The decreased level of O-GlcNAc modification in endothelial cells stimulated with LPS promotes partial activation of inflammatory signaling pathways, mainly involving ERK, p38MAPK, and STAT3, and affects the expression of inflammatory factors. AKT may be involved in the effect of LPS on the inhibition of O-GlcNAc modification.

Objective:To evaluate the clinical characteristics and prognosis of febrile convulsions (FS) in children during the epidemic period of novel coronavirus Omicron variant.Methods:A retrospective analysis was conducted on the clinical data of pediatric patients diagnosed with FS at Changsha Central Hospital Affiliated to University of South China from February 1, 2022 to January 31, 2023. The clinical characteristics and prognosis of FS patients caused by Omicron variant infection (observation group) were compared with those caused by non Omicron variant infection (control group).Results:A total of 131 cases in the observation group and 341 cases in the control group; The proportion of children aged 12-36 months in the observation group was lower than that in the control group ( P<0.05), and the proportion of children aged ≥60 months was higher than that in the control group ( P<0.05). Most of the FS in the observation group occurred within 24 hours of fever (128/131, 97.7%), with a statistically significant difference compared to the control group ( P<0.05), and most of them were generalized tonic clonic seizures (127/131, 96.9%), with only one seizure during the course of the disease (114/131, 87.0%), consistent with the characteristics of simple FS. The main clinical symptoms of FS patients in the observation group were upper respiratory tract infections (108/131, 82.5%), which were significantly higher than those in the control group (164/341, 48.1%), while the incidence of lower respiratory tract infections was lower than that in the control group ( P<0.05). 369 pediatric patients were followed up by phone or outpatient visits, with 98 cases in the observation group and 2 cases experiencing recurrence. There was no recurrence in the group aged ≥60 months; A total of 271 cases were followed up in the control group, with 9 cases experiencing recurrence. Conclusions:The number of children with FS caused by novel coronavirus Omicron variant has increased sharply, and the proportion of late onset FS patients has increased significantly. Most of them are upper respiratory tract infections. Convulsions usually occur within 24 hours of fever, and the prognosis is good.

Objective:To explore the value of laparoscopic and endoscopic cooperative surgery with the patient lying on supine position under general anesthesia in the operation of type I Mirizzi syndrome with choledocholithiasis.Methods:From Jan 2018 to Jan 2020, 53 cases of Mirizzi syndrome with choledocholithiasis undergoing laparoscopic and endoscopic cooperative surgery (preLC+ ERCP+ EST) at the Second Affiliated Hospital of Kunming Medical University were retrospectively analyzed.Results:53 patients successfully underwent LC without conversion to open surgery, and 2 patients failed in ERCP + EST attempt, with a success rate of 96.2%. One patient developed pancreas pseudocyst as a result of post-operative hemorrhagic necrotizing pancreatitis. Two patients suffered from chronic pancreatitis. Three patients complaining postoperative upper abdominal discomfort were finally diagnosed as stump cystic duct inflammation by MRCP, and no abnormalities were found in the follow-up of the remaining cases.Conclusion:Laparoscopic and endoscopic cooperative surgery in the treatment of patients with type I Mirizzi syndrome combined with choledocholithiasis is minimally invasive and effective.

Hepatic stellate cells(HSCs)are essential drivers of fibrogenesis.Inducing activated-HSC apoptosis is a promising strategy for treating hepatic fibrosis.18beta-glycyrrhetinic acid(18β-GA)is a natural com-pound that exists widely in herbal medicines,such as Glycyrrhiza uralensis Fisch,which is used for treating multiple liver diseases,especially in Asia.In the present study,we demonstrated that 18β-GA decreased hepatic fibrosis by inducing the apoptosis in activated HSCs.18β-GA inhibited the expression of α-smooth muscle actin and collagen type Ⅰ alpha-1.Using a chemoproteomic approach derived from activity-based protein profiling,together with cellular thermal shift assay and surface plasmon reso-nance,we found that 18β-GA covalently targeted peroxiredoxin 1(PRDX1)and peroxiredoxin 2(PRDX2)proteins via binding to active cysteine residues and thereby inhibited their enzymatic activities.18β-GA induced the elevation of reactive oxygen species(ROS),resulting in the apoptosis of activated HSCs.PRDX1 knockdown also led to ROS-mediated apoptosis in activated HSCs.Collectively,our findings revealed the target proteins and molecular mechanisms of 18β-GA in ameliorating hepatic fibrosis,highlighting the future development of 18β-GA as a novel therapeutic drug for hepatic fibrosis.

The composition of serum is extremely complex,which complicates the discovery of new pharmaco-dynamic biomarkers via serum proteome for disease prediction and diagnosis.Recently,nanoparticles have been reported to efficiently reduce the proportion of high-abundance proteins and enrich low-abundance proteins in serum.Here,we synthesized a silica-coated iron oxide nanoparticle and devel-oped a highly efficient and reproducible protein corona(PC)-based proteomic analysis strategy to improve the range of serum proteomic analysis.We identified 1,070 proteins with a median coefficient of variation of 12.56％using PC-based proteomic analysis,which was twice the number of proteins iden-tified by direct digestion.There were also more biological processes enriched with these proteins.We applied this strategy to identify more pharmacodynamic biomarkers on collagen-induced arthritis(CIA)rat model treated with methotrexate(MTX).The bioinformatic results indicated that 485 differentially expressed proteins(DEPs)were found in CIA rats,of which 323 DEPs recovered to near normal levels after treatment with MTX.This strategy can not only help enhance our understanding of the mechanisms of disease and drug action through serum proteomics studies,but also provide more pharmacodynamic biomarkers for disease prediction,diagnosis,and treatment.

Objective:A multi-center and large sample volume study was conducted on the verification and improvement of the early established criteria for intelligent routine urinalysis validation (including the microscopic review rules and manual validation rules, referred to as intelligent criteria for short), in order to improve the clinical application of this intelligent criteria.Methods:A total of 31 456 urine specimens were collected from the inpatients and outpatients in six hospitals in China, from March to September 2019. Firstly, 3105 specimens were analyzed for preliminary verification and improvement of the intelligent criteria based on the results of the microscopic examination and manual validation. Secondly, 28 351 specimens were used to verify the clinical application of the improved intelligent criteria. All samples were manually validated as reference.Results:The approval inconsistency rate of the manual validation rules in the original intelligent criteria was 8.59% (202/2 352), and the interception inconsistency rate was 8.84% (208/2 352). The false negative rate and the microscopic review rate of the microscopic review rules were similar to the previous results. Based on an in-depth analysis of big data and the discussions by senior technicians from eight hospitals, one microscopic review rules and four manual validation rules were added, meanwhile two manual validation rule was deleted. The manual validation standards were unified. Finally, the intelligent criteria was improved. Based on the improved intelligent criteria, for microscopic review rules, the false positive rate, false negative rate (misdiagnosis rate), and microscopic review rate did not change significantly, which were 14.72% (457/3 105), 4.06% (126/3 105), and 24.73% (768/3 105), respectively. The approval inconsistency rate and the interception inconsistency rate of manual validation rules were both reduced to 0; the total manual validation rate of the intelligent criteria was 50.89% (1 580/3 105), and the auto-validation rate was 49.11% (1 525/3 105). The large sample volume verification results were consistent with the preliminary verification results of the improved intelligent criteria.Conclusion:This multi-center and large sample volume study had shown that the improved intelligent criteria had better clinical performance.