Programmed death-ligand 1 (PD-L1 ),which is highly expressed in NSCLC,can be divided into two categories:membrane PD-L1 and soluble PD-L1 .PD-L1 participates in tumor immune escape through combining with its receptor.PD-L1 immune checkpoint inhibitors have entered the phase Ⅰ studies and showed a good application prospect.It is also found that the sensitivity of PD-L1 immune checkpoint inhibitors is strongly associated with the expression of PD-L1 in tumor.Therefore,PD-L1 can be used as a biomarker to pre-dict its curative effect.

ObjectiveTo analyze the change in the incidence of papillary thyroid carcinoma (PTC) and follicular thyroid carcinoma (FTC) in the past 27 years and to probe the relationship between iodine and these carcinomas. MethodsStatistical analysis was performed on data of thyroid diseases, thyroid malignant tumors, PTC , FTC and their complications in Department of Pathology from 1978 to 2004 in General Hospital of Tianjin Medical University with Run test、Cox-Stuart test, ANOVA, t or t′ test and Chi-square test. ResultsIn 27 years there were 231837 cases of biopsies (average 8587cases/year). The biopsies were performed in the thyroid diseases as a whole 6276 cases (2.70%),thyroidmalignanttumors644cases(0.28%), PTCs 465 cases (0.20%), and FTCs 80 cases (0.03%) in 27 years. Biopsies in thyroid diseases as a whole showed a descending tendency since 1982 but an ascending tendency after 1995. Biopsy in malignant thyroid tumor did not show a significant change, and biopsy in PTC showed an ascending tendency and that in FTC showed a descending tendency after 1995. The average age, both in male and female patients, at the time of diagnosis of PTC and FTC showed no significant change during 27 years, but the mean age of male at the time of diagnosis was older than that of female. The ratios of male to female in PTC and FTC were 1:3.31、1:2.81 respectively,and showed no significant change either. There were more biopsies with lymph node metastasis in PTC (33.76%) than those in FTC (10.00%) and more biopsies with blood vessel involved or nodular goiter in FTC (11.25% and 13.75%) than those in PTC (0.65% and 4.52%). Mean age of patients with PTC and nodular goiter (44.3 years old) was older than those without goiter (38.1 years old). The ratios of male to female with PTC and FTC with lymph node metastasis were higher than those without lymph node metastasis. ConclusionThe incidence of thyroid malignant tumors in 27 years has no significant change. The contrary tendency of PTC and FTC suggests that the decreasing malignant degree of carcinoma. It suggests that this area is not in high iodine condition and the change have no relationship with the iodine intake.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Emodin ; pharmacology ; Humans ; Neoplasms ; pathology ; Tumor Cells, Cultured

Objective To evaluate the features of the encapsulated papillary thyroid carcinoma (EPTC) by ultrasound and histopathology.Methods The EPTC were classified into the following two types based on the shape,characteristics of the border,size of the nodule,echogenicity,a hypoechoic halo and microcalcification by ultrasound features:papillary carcinoma (PC) type and follicular tumor (FT) type.Results Of all the 33 cases,21 cases were PC type and 12 cases were FT type.The histopathological result of PC type was papillary carcinoma.PC type had a jagged border,an irregular tumor shape with marked hypoechogenicity by ultrasound.PC type were composed of papillae by histopathology,completely surrounded by a fibrous capsule with tumor cells having the nuclear features of papillary thyroid carcinoma,densely interstitial fibrosis and microcalcification;FT type had a smooth border,a regular shape (spherical to oval),isoechogenicity and a hypoechoic halo by ultrasound.FT type were completely or significantly composed of follicles by histopathology,completely surrounded by a fibrous capsule with tumor cells having the nuclear features of papillary thyroid carcinoma.Hypoechoic halo were more frequently observed in FT type than in PC type.The nodule size of FT type(1.8-7.0 cm)was larger than that of PC type(0.8-5.2 cm).Fine and multiple strong echoes were characteristically present only in PC type.Conclusion The EPTC have characteristic features that are similar to those of the benign follicular thyroid tumor by ultrasound.

Objective To study the role of hydrogen sulfide (H2S) in the effect of SB203580 on proliferation and apoptosis of hepatic stellate cells and the effects of H2S on expressions of collagenⅠand collagenⅢmRNA in hepatic stel-late cells. Methods There were five groups of HSC-T6 cells in this study including control group (DMEM medium contain-ing10%fetal bovine serum), dimethyl sulfoxide (DMSO) group, sodium hydrosulfide (NaHS)group,SB203580 (SB)group and SB+NaHS group. MTT method was used to detect the cell proliferation and inhibition rate of HSC-T6 cells treated by SB203580 and H2S. The apoptotic rate of HSC-T6 cells was detected by flow cytometry with annexin V-FITC/PI double staining. RT-PCR was used to detect the expressions of collagenⅠand collagenⅢmRNA in HSC-T6. Results The apop-totic rate of HSC-T6 cells was significantly higher in SB group and SB+NaHS group than that of control group, and the rate was significantly higher in SB+NaHS group than that of SB group (P＜0.05). There was no significant difference in the apop-totic rate of HSC-T6 cells between DMSO and NaHS groups than that of control group. The expressions of collagenⅠand col-lagenⅢmRNA were found in five groups of cells. There was a higher expression of collagenⅠand collagenⅢmRNA in NaHS group than that of control group (P＜0.05). The expressions of collagenⅠand collagenⅢmRNA were significantly lower in SB group and SB+NaHS group than those of control group and NaHS group (P＜0.05). Conclusion H2S activated P38MAPK signal pathway. And P38MAPK was specifically blocked by SB203580 in HSC-T6 cells, which inhibited the cell proliferation stimulated by H2S and promoted the apoptosis.

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

Objective To investigate the meaning of expression of apoptosis related molecules NFKBp65 and p53 and GPX1-mRNA in patients with Keshan disease(KSD).Methods Sixteen chronic Keshan Disease patients were enrolled in KSD group according to electrocardiogram,chest X ray film and clinical examinations on 15,September in 2009,and 23 healthy people were included in control group from physical examination taken in The Second Affiliated Hospital of Xi'an Jiaotong University.Fresh blood(5 ml)was collected from antecubital vein of all subjects in the fasting state.Total mRNA and protein of blood sample were isolated using Trizol.GPX Assay Kit was used to detect GPX enzyme activity,and GPX1-mRNA expression was determined by SYBR Real-Time PCR.Meanwhile,expression of apoptosis related molecules NFKBp65 and p53 were determined by Western blot.Results GPX enzyme activity decreased significantly in KSD group[(108.61±14.10)U]compared with control group[(122.78±11.89)U,t=2.874,P<0.05],GPX1-mRNA level of KSD group(0.553±0.299)notably KSD group(0.802±0.057)compared with control group[(1.065±0.355),t=6.829,P<0.01].p53 increased in KSD group(1.604±0.191)compared with control group[(1.137±0.186),t=3.033,P<0.05].Conclusiom Decreased GPX1-mRNA expression may result in lower GPX enzyme activity of patients with KSD.Thus oxidative damage increases and cadioeyte apoptosis is activated by activating apoptosis signal pathway.

OBJECTIVETo study the effect of transforming growth factor-β activated kinase-1 (TAK1) gene silencing on the proliferation and apoptosis of Kasumi-1 cells induced by arsenic trioxide (As₂O₃).

METHODSAcute myeloid leukemia with t(8;21) cell line Kasumi-1 cells were treated with As₂O₃ or in combination with TAK1 siRNA interference technology. The experiment was divided into four groups: Kasumi-1 cells without any treatment, TAK1 specific siRNA transfection alone, Kasumi-1 cells treated with different concentration of As₂O₃, TAK1siRNA transfection combined with As₂O₃. CCK-8 was used to detect the cell viability. The expression of phosphorylated c-Jun N-terminal kinase (P-JNK) was determined by Western Blot. Cell apoptosis and growth were examined by morphological and colony formation assay.

RESULTSAfter Kasumi-1 cells were treated with As₂O₃, the rate of cell inhibition was concentration-dependent, and the 50% inhibitory concentration was 3.5 μmol/L. The highest expression level of P-JNK appeared in 30 minutes after cells were treated with As₂O₃. The apoptosis rates of Kasumi-1 cells without any treatment, TAK1 siRNA interference alone group, As₂O₃ alone group and the combined group were (5.02 ± 1.13)%, (6.18 ± 0.28)%, (48.33 ± 2.70)% and (86.07 ± 2.21)%; colony formation rates were (73.83 ± 2.78)%, (76.03 ± 1.46)%, (55.07 ± 1.50)% and (22.20 ± 1.15)%; apoptosis rate of TAK1 siRNA group and the untreated group has no significant difference (P = 0.052); colony formation rate between TAk1 siRNA group and the untreated group has no significant difference (P = 0.179), but the difference in other groups was significant (P = 0.000).

CONCLUSIONSilencing the expression of TAK1 can enhance the anti-proliferative and pro-apoptotic effect of As₂O₃ on Kasumi-1 cells, and its mechanism may be through the TAK1 downstream JNK signal pathway.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Line, Tumor ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Leukemia, Myeloid, Acute ; enzymology ; pathology ; MAP Kinase Kinase Kinases ; genetics ; metabolism ; Oxides ; pharmacology ; RNA Interference ; RNA, Small Interfering ; genetics ; Signal Transduction

Objective To study the clinical significance of fecal smear examination on diagnosing intestinal flora imbalance in infantile diarrhea.Methods A sterile cotton swab was used to spread a layer of fresh feces quantum satis from a sterile container on a clean slide;the smear was fixed and stained with Gram′s methods after it was air-dried,then the specimen was observed with a microscopy(field lens 100 ? eye lens 10) and recorded.Results In the acute diarrhea group(40 cases),the distribution feature of 3 floras on the fecal smears:≥50% Gram-negative bacilli in 6 cases(15%),≥50% Gram-positive cocci in 30 cases(75%) and ≥68% Gram-positive bacilli in 4 cases(10%).In the delayed and chronic diarrhea group(62 cases),the distribution feature of 3 floras on the fecal smears:≥50% Gram-negative bacilli in 7 cases(11.29%),≥50% Gram-positive cocci in 44 cases(70.97%) and ≥68% Gram-positive bacilli in 6 cases(9.68%).In the normal control group(32 cases),the distribution feature of 3 floras on the fecal smears:≥50% Gram-negative bacilli in 1 case(3.13%),≥50% Gram-positive cocci in 1 case(3.13%),and ≥68% Gram-positive bacilli in 17 cases(53.13%).For the distribution of 3 floras in the 3 groups,chi-squared test was performed,and the results showed that the difference was significant in Gram-positive cocci(?~2=47.76 P0.05).Conclusions Acute,delayed or chronic diarrhea can lead to flora imbalance.In order to timely and rapidly know the flora imbalance in children with diarrhea,clinically the simple,easily operated and practical smear staining method shall be widely applied.

Abstract: Objective　To investigate the clinical characteristics and incidence of Brucella encephalitis and meningitis in children. Methods We report the clinical data of a child with Brucella melitensis meningitis in children, and summarize the incidence, diagnosis methods and treatment of Brucella encephalitis or meningitis in children, taking into account the relevant domestic and foreign literature from January 2014 to December 2020. Results　A 4-year-old girl was admitted to the hospital with status epilepticus on March 15, 2021 because of interrupted right limb numbness for 16 hours and convulsions for 2 hours. She had 2 non-febrile convulsions three months before admission and was diagnosed with epilepsy. This incident was acute, accompanied by low fever, with epilepsy as the main manifestation. Cerebrospinal fluid test suggested central nervous system infection, but the nature of infection could not be determined by routine and biochemistry of cerebrospinal fluid.The cerebrospinal fluid next generation sequencing confirmed that the pathogen of the infection was B. melitensis, which was further verified by the peripheral blood antibody test. After effective antibiotics combined with a full course of treatment, the patient recovered after six months of treatment. A total of 60 articles were retrieved in the database, including 29 in Chinese. During this period, a total of 7 cases of brucellosis in children with nervous system involvement were reported, one of which was a case report, and the other 6 cases were mentioned in the comprehensive analysis of children with brucellosis. Conclusions　Brucella encephalitis or meningitis in children has a low incidence and various clinical features, which are easy to be misdiagnosed or missed.