Objective:To explore the alteration of structural network, cognitive scores in patients with anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis, as well as the relationship between cognitive impairment and changes of structural networks in patients with NMDAR encephalitis.Methods:This study was a retrospective study. A total of 39 patients with anti-NMDAR encephalitis were recruited as the autoimmune encephalitis group (AE group) from the First Affiliated Hospital of Chongqing Medical University between September 2012 to December 2019, and 32 healthy volunteers with normal results of routine head MR examinations and no history of central nervous system diseases were recruited as the health control group (HC group). There were 16 males and 23 females, aged from 13 to 66 (34±15) years, with duration of disease from 11 to 110 (31±20) days in AE group, and there were 16 males and 23 females, aged from 13 to 66 (34±15) years in HC group. All subjects underwent diffusion tensor imaging (DTI) scan and cognitive function evaluation. The brain structural networks of two groups were constructed by deterministic fiber tracking techniques, and the differences of global topological properties [clustering coefficient (C p), shortest path length (L p), local efficiency (E loc), global efficiency (E glob), normalized C p (γ), normalized L p (λ), small-worldness (σ)] and local topological properties between two groups were analyzed by the graph theory approch. The correlations between characteristics of brain structural networks and cognitive function scores were further analyzed. Results:There was no significant difference in age and gender distribution between the AE group and HC group ( P>0.05). The C p [0.005(0.004, 0.007)], γ (1.76±0.13), λ (0.51±0.03) and σ value (1.57±0.13) of AE group were decreased when compared with HC group [the values were 0.007(0.004,0.017), 2.13±0.63, 0.55±0.06 and 1.73±0.36 each] ( Z=-939.00, t=-3.58, t=-4.16, t=-2.58, P<0.05). Compared with HC group, nodal efficiencies in the left middle frontal gyrus (orbital part), left and right supplementary motor areas, left olfactory cortex, left gyrus rectus, bilateral insula, left postcentral gyrus, left paracentral lobule and right heschl gyrus were changed ( P<0.05). There were five identical hub regions which contains the left middle occipital gyrus, bilateral supplementary motor areas and precuneus in both groups. However, in the AE group, three hub regions of the left middle occipital gyrus and bilateral middle temporal gyrus were reduced, and the left precentral gyrus was increased as hub region. The nodal efficiencies of the left supplementary motor areas ( r=0.393, P=0.013), right supplementary motor areas ( r=0.384, P=0.016) and left paracentral lobule ( r=0.356, P=0.026) were positively correlated with the montreal cognitive assessment scores. Conclusion:The white matter is extensively impaired in anti-NMDAR encephalitis patients and the changes of topological properties in several brain regions are correlated with cognitive decline.

Objective: To establish a rapid GeXP-based multiplex reverse transcription-PCR assay (GeXP assay) for simultaneous detection of 5 subtypes of diarrheogenic Escherichia coli. Methods: Specific primers were designed according to reserved sequences of 12 virulence genes in enterotoxigenic E. coli (ETEC), enterinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC) and enterohemorrhagic E. coli (ETEC), and PCR amplification was performed with a single pair of primers to validate the specificity of PCR assay with a single template and a single pair of primers. The specificity of the GeXP assay was evaluated with the genomic DNA of 5 subtypes of diarrheogenic E. coli as the template in a mixture of 12 pairs of primers, and the sensitivity of the GeXP assay was evaluated with the mixed suspensions of 5 subtypes of diarrheogenic E. coli at concentrations of 106, 105, 104 and 103 CFU/mL as the template. Foods purchased from supermarkets and agricultural retail markets were prepared into 34 spiked samples, and 5 subtypes of diarrheogenic E. coli were detected using the GeXP assay and compared with the fluorescent real-time multiple PCR assay. Results: The sizes of sth, pic, bfpB, astA, lt, escV, aggR, stx1, uidA, invE, stx2 and stp genes amplification products were consistent with expected sizes using a single template and a single pair of primers, with a fluorescent signal intensity of more than 25 000 A.U. The sizes of the GeXP assay amplification products of 12 virulence genes in 5 subtypes of diarrheogenic E. coli were consistent with expected sizes, with a high specificity. If the concentration of the mixed suspensions of 5 subtypes of diarrheogenic E. coli was 103 CFU/mL, the GeXP assay was effective for simultaneous detection of 12 virulence genes, with a high fluorescent signal intensity, consistent repeated detection results and a less than 10% coefficient of variation. The GeXP assay detected 3 ETEC isolates, 12 EAEC isolates, one EIEC isolate, one EPEC isolate and one EHEC isolate among the 34 spiked samples, which was in agreement with the detection of 5 subtypes of diarrheogenic E. coli with commercial fluorescent real-time multiple PCR assay kits. Conclusions A GeXP assay has been successfully established for simultaneous detection of 12 virulence genes in diarrheogenic E. coli, which is effective for clinical differential diagnosis and epidemiological surveys of diarrheogenic E. coli.

Aim To investigate the role of phospho-lipase C epsilon 1 ( PLCE1 ) in modulating the apoptot-ic mechanism in esophageal cancer ( Eca ) cells. Methods Eca cell lines, OE33 and CP-C cells were cultured to assess the expression of PLCE1 . siRNA suppress expression of PLCE1 . The expression of PLCE1 and p53 was evaluated by quantitative real time PCR and Western blot. Methylation analyses of p53 were performed by bisulfite conversion of genomic DNA. Apoptosis was assessed by flow cytometry. Results OE33 and CP-C cells expressed high levels of PLCE1 . Knockdown of PLCE1 markedly increased the expression and hypomethylation of p53 , and in-creased the frequency of apoptosis. Conclusion PLCE1 suppresses apoptosis of Eca cells via promoting p53 promoter methylation and inhibiting expression of p53 .

AIM:To investigate the role of phospholipase C epsilon 1 ( PLCE1 ) in modulating the apoptotic mechanism in lung adenocarcinoma A 549 cells.METHODS:PLCE1 inhibitor U-73122 was used to suppress the expres-sion of PLCE1.The expression of PLCE1 and p53 in A549 cells was evaluated by quantitative real-time PCR and Western blotting.Apoptosis was assessed by flow cytometry .RESULTS:A549 cells expressed high level of PLCE1 and low level of p53.Inhibition of PLCE1 markedly increased the expression of p 53, and increased the apoptosis of A 549 cells.CON-CLUSION:PLCE1 suppresses apoptosis of A549 cells via inhibiting the expression of p53.

Aim To investigate the effects of airway epithelial cell-derived insulin-like growth factor-1 (IGF1) on CD8 +T cell polarization. Methods Hu-man airway epithelial cell line, RPMI2650 cells, was cultured in the presence of a mice allergen, Der p1, for 72 h. IGF1 expression was checked with quantita-tive RT-PCR and Western blot. Der p1-primed RP-MI2650 cells, recombinant IGF1 and anti-IGF1 anti-body was cocultured respectively with CD8 + T cells, which were activated by anti-CD3/CD8 Ab. Apoptotic cells frequency was calculated with flow cytometry. The alteration of p53 gene hypermethylation in CD8 + T cells elicited by Der p1-primed airway epithelial cell and IGF1 was plotted. Results Both mRNA(23. 1%± 5. 2% vs 5. 2% ± 2. 3%, P < 0. 01 ) and protein (33. 4 ± 6. 4 vs 9. 2 ± 4. 6, P <0. 01 ) expression of IGF1 in RPMI2650 cells markedly increased after ex-posure to Der p1 . The increase of apoptotic CD3/CD28 Ab-activated CD8 + T cells was abolished by the pres-ence of Derp1-primed epithelial cells ( 41. 7% ± 8. 2%vs 5. 2% ± 1. 8%, P <0. 01 ) . The results were con-firmed by the addition of recombinant IGF1 . Anti-IGF1 antibody abolished the effect of the epithelial cells. Derp1-primed epithelial cells inhibited p53 gene mR-NA( 29. 1% ± 5. 9% vs 16. 2% ± 4. 3%, P <0. 01 ) and protein ( 63. 3 ± 8. 9 vs 26. 9 ± 5. 6 , P <0. 01 ) ex-pression. Anti-IGF1 antibody abolished the effect. Re-combinant IGF1 promoted CD8 + T cells′p53 gene hy-permethylation. Conclusion Der p1 induces RP-MI2650 cells to produce IGF1 , and this factor prevents CD8 + T cell apoptosis by inducing p53 gene hyperm-ethylation.

ObjectiveTo explore the application value of thoracoscopic pleural lavage for empyema after liver transplantation (LT).MethodsClinical data of 5 patients who underwent thoracoscopic pleural lavage for empyema after LT in the Third Afifliated Hospital of Sun Yat-sen University from October 2002 to October 2013 were analyzed retrospectively. All 5 patients were males with the age ranging from 42 to 56 years old and the median of 51 years old. The informed consents of all patients were obtained and the local ethical committee approval had been received. The patients received thoracoscopic pleural lavage for empyema under endotracheal general anesthesia. The thoracoscope was introduced in the seventh intercostal space on the mid-axillary line. The aspirator was inserted in the third intercostal space on the anterior line to drain the pus. And the pleural ifberboard was peeled off. The intraoperative situation and perioperative recovery of the patients were observed.ResultsFive patients received successful operations and recovered and were discharged from hospital. The median length of operation was 2(1-3) h. The length of respirator assisted ventilation was 26(6-42) h. The intraoperative blood loss was 600(300-1 000) ml. The length of indwelling chest drainage tube was 4(2-6) h. The volume of chest drainage was 680(350-1 200) ml. No mortality, empyema recurrence and other serious complications were observed. The postoperative hospital stay was 7(5-11) d.ConclusionThoracoscopic pleural lavage for empyema after LT is a safe and effective treatment.

OBJECTIVETo investigate the cytotoxicity of normal CD8(+) T lymphocytes retrovirally transduced with WT1 peptide-specific T-cell receptor (TCR) genes against human lung cancer cells.

METHODSHLA-A*2402-restricted and WT1 peptide-specific TCR-α/β genes were cloned from a cytotoxic T lymphocyte clone and inserted into a retroviral TCR expression vector. The cytotoxicity of normal peripheral CD8⁺ T cells transduced with the WT1-TCR genes against human lung cancer cells was evaluated using a standard ⁵¹Cr release assay.

RESULTSThe WT1-TCR gene-modified T cells recognized the peptide-pulsed target cells but not the non-pulsed cells. TCR-redirected CD8⁺ T cells lysed WT1-overexpressing human lung cancer cells in an HLA-A*2402-restricted manner, but did not kill normal cells positively expressing HLA-A*2402.

CONCLUSIONThese data demonstrate the feasibility of adoptive immunotherapy with TCR-redirected T cell for the treatment of lung cancer.

CD8-Positive T-Lymphocytes ; cytology ; Cell Line, Tumor ; Genes, T-Cell Receptor ; Humans ; Immunotherapy, Adoptive ; Lung Neoplasms ; pathology ; Peptides ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; Retroviridae ; T-Lymphocytes, Cytotoxic ; cytology ; Transduction, Genetic ; WT1 Proteins ; genetics

Aim Toinvestigatewhetherepidermal growth factor receptor ( EGFR )-containing exosomes could induce tumor-specific regulatory T cells, and the effects of those T cells on tumor protein-specific CD8+Tcells.Methods TheexosomeswithEGFRwerepu-rified from NSCLC tumor, which modulated tolerogenic property of DCs. Then the induced TolDCs generated tumor-specific Tregs, with which the tumor protein-specific CD8 + T cells were suppressed. Results 80%exosomeswereEGFRpositivefromLCpatients while less than 2% exosomes were EGFR positive from control lung tissue. After exposed to the exosomes in the culture for 7 days, the IDO+ DCs proportion was much higher than that in control group ( 80. 8% ± 3. 2% vs 65. 6% ± 6. 4%, P <0. 05 ) . The induced Tregs was also higher ( 24. 1% ± 5. 2% vs 4. 2% ± 2. 3%,P<0. 01 ) , which suppressed the proliferation of CD8+ T cells(5. 4% ± 0. 2% vs 86. 7% ± 9. 3%, P<0.01).Conclusion Thepurifiedexosomesin-duce tolerogenic DCs. Coculture of the tolerogenic DCs and Th0 cells generates the tumor antigen-specific reg-ulatory T cells. The Tregs could suppress the tumor an-tigen specific CD8+ T cells.

Aim To investigate the activation of prote-ase-activated receptor 2 ( PAR2 ) in regulation of the expression of epidermal growth factor receptor ( EGFR) and apoptosis of lung cancer ( LC) cells. Methods LC cells A549 and its EGFR-silenced counterpart were incubated in the medium added with tryptase. Quanti-tative RT-PCR and Western blotting were used to de-tect the expression of EGFR in A 5 4 9 cells . The apop-tosis and Bax/Bcl-xL of cells were also recorded. Re-sults Treating A549 cells with tryptase in the culture for 48 hrs resulted in a marked increase in the expres-sion of EGFR in A549 cells. Marked decreases in a tryptase dose-dependent manner in apoptotic A549 cells were detected in the presence of tryptase. Expo-sure to tryptase markedly decreased the levels of Bax and increased the levels of Bcl-xL in A549 cells, which were not shown in EGFR-deficient A549 cells. Conclusion Tryptase can increase the expression of EGFR in LC cell line, A549 cells, which protects A549 cells from apoptosis, increases Bcl-xL, and sup-presses Bax in A549 cells.

OBJECTIVEMetabonomics researches of Scutellariae Radix interfering pyretic pulmonary syndrome had been done, to determine the specific biomarkers of pyretic pulmonary syndrome, and to approach the mechanism that Scutellariae Radix interfered the variation of these biomarkers.

METHODMetabonomics technique, UPLC-Q-TOF/MS analytical means and PCA statistical methods were utilized to investigate the trajectory change and inter-relationship of urinary metabolome of rats treated differently.

RESULTSix specific biomarkers were determined which could represent Streptococcus pneumoniae-induced pyretic pulmonary syndrome in rats. Scutellariae Radix could significantly adjust the ascended biomarkers to the normal level. Meanwhile two of these biomarkers were identified as Delta-12-prostaglandin J2 and indoxyl sulfate.

CONCLUSIONThere was a good therapeutic function of Scutellariae Radix on pyretic pulmonary syndrome, which was elucidated on the metabolic aspects. There was also a correlationship between the mechanism of Scutellariae Radix curing pyretic pulmonary syndrome and the six specific biomarkers.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Lung Diseases ; drug therapy ; metabolism ; Male ; Pneumococcal Infections ; drug therapy ; metabolism ; Rats ; Rats, Wistar ; Scutellaria baicalensis ; chemistry