OBJECTIVEThe objective of this article was to review the current advances in diagnostic methods for Acanthamoeba keratitis (AK).

DATA SOURCESData used in this review were retrieved from PubMed (1970-2013). The terms "Acanthamoeba keratitis" and "diagnosis" were used for the literature search.

STUDY SELECTIONData from published articles regarding AK and diagnosis in clinical trials were identified and reviewed.

RESULTSThe diagnostic methods for the eight species implicated in AK were reviewed. Among all diagnostic procedures, corneal scraping and smear examination was an essential diagnostic method. Polymerase chain reaction was the most sensitive and accurate detection method. Culturing of Acanthamoeba was a reliable method for final diagnosis of AK. Confocal microscopy to detect Acanthamoeba was also effective, without any invasive procedure, and was helpful in the early diagnosis of AK.

CONCLUSIONClinically, conjunction of various diagnostic methods to diagnose AK was necessary.

OBJECTIVE: To explore the influence of clinical practice on mental health of male nursing college students,and to investigate the effects of psychological intervention. METHODS: Using the cluster sampling method,408 clinical practice male nursing students from 5 medical colleges in Jilin Province were randomly divided into intervention group and control group. The male nursing students were investigated with Chinese Perceived Stress Scale(CPSS),M to R sections of Cornell Medical Index(CMI) and Symptom Checklist 90(SCL-90). After the first questionnaire survey in the 9 weeks of clinical practice(before psychological intervention),male nursing students in intervention group received psychological intervention,while those in control group received no action. The investigation were then performed in the 20th(11 weeks of intervention) and 30 th weeks(21 weeks of intervention) of clinical practice. RESULTS: In the 11 and 21 weeks of intervention,the positive rates of CPSS,M-R and mental symptoms in intervention group were lower than those of timematched control group(P < 0. 01),and were significantly lower than those before intervention(P < 0. 01). Before intervention,the scores of CPSS,M-R and SCL-90 in two groups had no statistical significance(P > 0. 05). After the 11 and 21 weeks of intervention,the scores of CPSS,M-R and SCL-90 in intervention group were lower than those of control group(P < 0. 01). With the increasing time of intervention,the scores of CPSS and M-R in intervention group decreased(P < 0. 01). After the 11 and 21 weeks of intervention,the scores of SCL-90 in intervention group were lower than those before intervention(P < 0. 01). CONCLUSION: The results suggest that clinical practice is a stress for the mental health of male nursing college students. Psychological supporting system can be used to improve students' psychological health.

Objective To investigate the efficacy and safety of endoscopic retrograde cholangiopancreatography (ERCP)+SpyGlass system versus percutaneous transhepatic gallbladder drainage (PTGD)+ERCP in the treatment of acute cholecystitis secondary to choledocholithiasis. Methods A retrospective analysis was performed for the clinical data of the patients with acute cholecystitis secondary to choledocholithiasis who were treated in Department of Gastroenterology, Jilin City People's Hospital, from December 2019 to September 2021, among whom there were 23 patients in the ERCP+SpyGlass group and 19 patients in the PTGD+ERCP group. The two groups were compared in terms of the indicators such as surgical technical success, surgical operation time, surgical clinical success, postoperative recovery, length of hospital stay, and complications. The two-independent-samples t test was used for comparison of normally distributed continuous data between groups, and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between groups; the chi- square test or the Fisher's exact test was used for comparison of categorical data between groups. Results Compared with the PTGD+ERCP group, the ERCP+SpyGlass group had a significant reduction in C-reactive protein after surgery ( Z =2.999, P =0.003). There were no significant differences between the two groups in technical success rate ( χ 2 =1.735, P =0.188), clinical success rate ( χ 2 =0.846, P =0.358), total time of operation ( t =1.667, P = 0.113), white blood cell count on day 1 after surgery ( t =1.075, P = 0.289), length of postoperative hospital stay ( t =1.560, P =0.127), and incidence rate of complications (all P > 0.05). Conclusion In the treatment of acute cholecystitis secondary to choledocholithiasis, the ERCP+SpyGlass system has a comparable clinical effect to PTGD+ERCP and is safe and effective, without increasing surgery-related adverse events and risks, and it can also solve the problems of the biliary tract and the gallbladder at one time through natural orifices, with no scars on body surface and convenient postoperative nursing. Therefore, it holds promise for clinical application.

ObjectiveTo investigate the safety and efficacy of endoscopic retrograde cholangiopancreatography （ERCP） combined with electrohydraulic lithotripsy under the direct view of eyeMax biliary-pancreatic imaging system in the treatment of difficult choledocholithiasis. MethodsA retrospective analysis was performed for the clinical data of 12 patients with difficult choledocholithiasis who underwent ERCP and electrohydraulic lithotripsy under the direct view of eyeMax biliary-pancreatic imaging system in Department of Gastroenterology， Jilin People’s Hospital， from May to November 2022. The clinical effect of lithotripsy and lithotomy was observed， and postoperative complications and time of surgical operation were assessed. ResultsAmong the 12 patients， 11 （91.67%） were successfully treated by electrohydraulic lithotripsy under direct view， 9 （75.00%） achieved first-attempt success in lithotripsy， and 11 （91.67%） had complete removal of calculi； 1 patient was found to have stenosis of the bile ducts caused by multiple biliary tract surgeries， and grade Ⅱ intrahepatic bile duct stones above the sites of stenosis were removed under direct view， but there were still residues of grade Ⅲ intrahepatic bile duct stones， which led to the fact that complete calculus removal was not achieved. The mean time of ERCP operation was 91.3±26.2 minutes， including a time of 41.8±22.2 minutes for energy lithotripsy. There were 2 cases of postoperative biliary tract infection which were improved after anti-infective therapy， 2 cases of hyperamylasemia which were not given special treatment， and 3 cases of mild pancreatitis which were improved after symptomatic medication， and there were no complications such as bleeding and perforation. ConclusionERCP combined with electrohydraulic lithotripsy under the direct view of eyeMax biliary-pancreatic imaging system is safe， effective， and feasible in the treatment of difficult choledocholithiasis.

OBJECTIVE To study the immunomodulatory effect of Guipi pills on D-galactose（D-gal）-induced aging mice. METHODS The immune-related targets and related pathways for Guipi pills to exert immune effects were screened by network pharmacology and verified through pharmacodynamic experiments. Totally 105 male ICR mice were randomly divided into blank control （CON） group， model control （MOD） group， positive control （POS） group， Guipi pills low-dose （GD） group and Guipi pills high-dose （GG） group. Except for the CON group， other groups were subcutaneously injected with 400 mg/kg D-gal to induce the aging model； CON group and MOD group were given distilled water， POS group was given 300 mg/kg pidotimod oral solution intragastrically， GD group and GG group were given Guipi pills 300， 600 mg/kg intragastrically， once a day， for 8 weeks. After medication， the serum and spleen were collected， and the contents of interleukin 2 （IL-2）， IL-4， IL-6 and tumor necrosis factor α （TNF-α）， and the contents of immunoglobulin G （IgG）， IgM and IgA were detected. The spleen index was calculated and the histopathological changes in the spleen were observed. The activities of superoxide dismutase （SOD）， glutathione peroxidase （GSH- Px） and malondialdehyde （MDA）， and the content of 8-hydroxy-2 deoxyguanosine （8-OHdG） in spleen were detected； the expression of TNF/phosphoinositide-3-kinase-threonine protein kinase （PI3k-Akt）-related proteins in spleen was detected except for POS group. RESULTS The results of network pharmacology showed that TNF， IL-6 and Akt1 were core targets. The results of pharmacodynamic study showed that compared with MOD group， the contents of IL-2， IL-4， IgG， IgM and IgA were increased significantly in Guipi pills groups， while the contents of TNF-α and IL-6 were decreased significantly； the spleen index was increased significantly （P＜0.05 or P＜0.01）. The phenomenon of diffuse proliferation of lymphocytes was improved， the spleen cells were closely arranged， and the line between the white pulp and red pulp was clear. The activities of SOD and GSH-Px in spleen were increased significantly， while the activity of MDA， the content of 8-OHdG， and the protein expressions of TNF-α， PI3K and p-Akt were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS Guipi pills can regulate the immune function of D-gal-induced aging mice， which is related to regulating the TNF/PI3k-Akt pathway， thereby reducing oxidative stress damage in spleen tissue of mice， and regulating protein expressions of TNF-α， PI3K and p-Akt.

The evolution, structure and antigenic epitopes prediction of Rana dybowskii antimicrobial peptide dybowskin-1ST were carried out using bioinformatics software available online. Its antibacterial mechanism and structural properties were analyzed, and its activity was verified by applying wound healing assay in mice and bacteriostatic assay in vitro. This provides the theoretical basis for the improvement of parental peptide and the development of novel derivative peptides. The software MEGA_X were used to conduct homology alignment and to construct a phylogenetic tree. The online software ProtParam, ProtScale, PeptideCutter, signal, TMHMM Server were respectively used to predict the physicochemical parameters, hydrophilia/hydrophobicity, shear sites, signal peptides, and transmembrane domains of dybowskin-1ST. The online software SOPMA, Jpred4, DNAstar Protean were used to predict the secondary structure of dybowskin-1ST, and SWISS-MODEL, I-TASSER were used to predict the tertiary structure. ABCpred and SYFPEITHI were respectively used to predict its B-and T-cell epitopes. The effect of dybowskin-1ST on the wound healing was observed on experimental mice. Kirby-Bauer method and dilution method were used to determine the bacteriostatic activity of dybowskin-1ST. The dybowskin-1ST consists of 59 amino acid residues, of which leucine accounts for 16.9%, with a molecular formula of C₃₁₈H₅₁₀N₈₀O₉₃S₂. Its theoretical isoelectric point is 5.10 and the charge is -2. The dybowskin-1ST and dybowskin-1CDYa are closely related phylogenetically. The secondary structure of dybowskin-1ST predicted by the three methods were similar, which consisted of α-helix (44.07%), extended strand (16.95%), β-turns (3.39%), and random coil (35.39%). The prediction of tertiary structure showed that dybowskin-1ST was mainly composed of α-helix, and it was regarded as a hydrophilic protein with signal peptide sequence. Subcellular localization analysis showed that the probability of secreting the mitochondrial targeted peptides was 0.944. Dybowskin-1ST is an extracellular protein with no transmembrane structure region, but contains seven phosphorylation sites, three T-cell epitopes and eight B-cell epitopes. The dybowskin-1ST promoted wound healing and effectively inhibited the growth of Escherichia coli and Staphylococcus aureus. However, it had limited antibacterial activity against fungi and drug-resistant bacteria. Although the structure of dybowskin-1ST is rich in α-helix, the verification experiments showed that its antibacterial ability needs to be enhanced. The reason may be that it is a negatively charged and hydrophilic protein, and amino acid modification with the aim of increasing the number of positive charges and changing the hydrophobicity may be used to obtain derived peptides with enhanced activity.
Amino Acid Sequence ; Animals ; Mice ; Phylogeny ; Pore Forming Cytotoxic Proteins ; Protein Structure, Secondary ; Ranidae

Objective To investigate the effects of hypoxia inducible factor-2?(HIF-2?) on the proliferation and invasion of hepatocellular carcinoma cell line SMMC-7721 under the anoxia condition.Methods Lipofectamine-mediated HIF-2? oligodeoxynucleotide(ODN) was transfected into SMMC-7721 cells and the expressions of HIF-2? mRNA and protein were detected to evaluate the transfection.The proliferative and adhesive activity was determined by methyl thiazolyl tetrazolium(MTT) method and the invasive activity was determined by a transwell cell culture chamber method.Results The expressions of HIF-2? mRNA and protein were inhibited significantly by HIF-2? antisense oligodeoxyribonucleotide(ASODN)(P0.05).Conclusion The blockage of HIF-2? expression inhibits the adhesive and invasive activity of SMMC-7721cells,and has no evident impact on their proliferation.

Rapid reduction of postprandial blood glucose is very beneficial to diabetics. In order to shorten the onset time of recombinant insulin, the cone snail insulin G1 (cI G1) of Conus geographus was studied. First, the nucleotide sequence of recombinant cone snail proinsulin G1 (cPI G1) was designed and synthesized according to the genes of human proinsulin (hPI) and cPI G1. The codon was optimized according to Escherichia coli (E. coli) codon usage frequency. Then, the plasmid pET22b(+)-cPI G1 was constructed and the recombinant cPI G1 was expressed in E. coli BL21(DE3) host strain. The recombinant cPI G1 was then purified and cleaved specially by trypsin to generate the recombinant cI G1, and its potency is 25.9 IU/mg. Fasting blood glucose test (FBGT) and oral glucose tolerance test (OGTT) suggested that the recombinant cI G1 could rapidly reduce blood glucose in normal and streptozotocin (STZ)-induced diabetic mice, but only for a short duration. This study provides a technical reference for the development of recombinant fast-acting insulin.
Animals ; Conus Snail ; Diabetes Mellitus, Experimental ; Escherichia coli ; Humans ; Hypoglycemic Agents ; Insulin ; Mice

Objective To observe the relationship between the expression of GFAP and iNOS the post-traumatic intervals after experimental cerebral contusion,in order to attempt to find out a method of timing wound age of brain contusion in the forensic prediction of injury time.Methods After setting up an model of experimental focal cerebral contusion in rats,immunohistochemical method was applied to observing the changes of inducible nitric oxide synthase (iNOS) and glial fibrillary acidic protein (GFAP) staining combined with histology method in rats.The immunostaining results were measured quantitatively with computer imaging analysis system.Results (1) At 3h after injury,GFAP-positive astrocytes around the wounded area were definitely detected.The intensity and the area of GFAP-positive cells increased following the post-traumatic intervals.The intensity peaked dually at 1d and 5d,respectively,followed by reduction at 3d,which revealed two-peak waveforms,The area of GFAP positive cells enhanced significantly at 3h and gradually increased with prolonging survival time,and reached a peak at 7d after injury.(2) At 12h after injury,the iNOS isoform was present and iNOS positive staining cells were noted.The intensity and the area of iNOS positive cells increased following the post-traumatic intervals.The area of positive reached maximal level at 1-3days,gradually subsided after 5d,and maintain a higher lever up to 7d post-injury;but the intensity of individual positive cell gradually increased with prolonging survival time,reached a peak at 5d and gradually subsided after 5d,but the level was still higher than the initial interval.Conclusions Expression of GFAP and iNOS is correlated with post-traumatic intervals after cerebral contusion in rats,suggesting that the expression of iNOS and GFAP may be served as the markers for timing of brain contusion in forensic practice.