Objective To investigate the effect of thalidomide on the proliferation of as well as the expression and secretion of vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) by the human keratinocyte cell line HaCaT.Methods Cultured HaCaT cells were divided into several groups to be treated with dimethyl sulfoxide (negative control group) and various concentrations (0.01nmol/L-100 μmol/L) of thalidomide (experimental groups) respectively for 20 to 24 hours.Subsequently,water soluble tetrazolium-1(WST-1) assay was performed to estimate cellular proliferation,real time quantitative PCR to detect the mRNA expression of VEGF and TNF-α in HaCaT cells,and enzyme-linked immunosorbent assay (ELISA) to quantify the protein expressions of VEGF and TNF-α in the culture supernatants of HaCaT cells.Statistical analysis was done by one-way analysis of variance with least significant difference post hoc test.Results The survival rate of HaCaT cells was 74.3％,82.9％ and 90.8％ after 24-hour treatment with thalidomide of 100,10 and 1 μmol/L respectively,significantly lower than that in the negative control group (100％,all P ＜0.01).A significant decrease was induced in the mRNA expression (0.439-to 0.634-fold change,all P ＜0.01) and supematant level ((0.587-to 0.923-fold change,P ＜0.05) of VEGF in HaCaT cells by thalidomide of 0.01-100 nmol/L,as well as in the mRNA expression (0.493-to 0.587-fold change,P ＜0.05) and supernatant level (0.408-to 0.617-fold change,P ＜0.01) of TNF-α by thalidomide of 0.1-100 nmol/L.Conclusion Within a certain range of concentration,thalidomide could suppress the proliferation of,as well as the expression and secretion of VEGF and TNF-α by,HaCaT cells.

Objective To evaluate the relationship of serum levels of interleukin (IL)-36α,IL-36β and IL-36γas well as their receptor antagonist IL-36Ra with disease severity in patients with psoriasis.Methods Venous blood samples were collected from 45 patients with generalized pustular psoriasis (GPP),34 patients with psoriasis vulgaris (PV),and 37 healthy human controls.Enzyme-linked immunosorbent assay (ELISA) was performed to determine the serum levels of IL-36α,IL-36β,IL-36γ and IL-36Ra.Nonparametric Mann-Whitney test was conducted to compare the levels of IL-36 and IL-36Ra among these groups,and Spearman's rank correlation analysis to assess the relationship between the serum levels of IL-36 and IL-36Ra and disease severity.Results No statistical difference was observed in the serum levels of IL-36 or IL-36Ra among the patients with GPP,patients with PV,and healthy human controls.The serum levels of IL-36β and IL-36γ (given as the median ± interquartile range) were significantly higher in 27 patients with GPP during episodes of pustules ((12.101 ± 11.315) ng/L and (34.541 ± 15.580) ng/L respectively) and in 7 patients with severe GPP ((11.218 ± 9.318) ng/L and (38.536 ± 17.332) ng/L respectively) than in the healthy human controls ((5.355 ± 9.020) ng/L and (23.052 ± 22.410) ng/L respectively,P ＜ 0.05 or 0.01).The serum level of IL-36γwas positively correlated with that of IL-36β in patients with GPP,patients with PV,and the healthy human controls (r =0.85,0.86,0.91,respectively,all P ＜ 0.01),and both IL-36β and IL-36γserum levels were lowly and positively correlated with the severity of GPP (r =0.33,0.41,respectively,both P ＜ 0.05).A positive correlation was also observed between the serum level of IL-36β and psoriasis area and severity index (PASI) scores in patients with PV (r =0.54,P ＜ 0.01).Conclusions The serum levels of IL-363 and IL-36γ are lowly and positively correlated with disease severity in patients with GPP,suggesting that IL-36β and IL-36γplay an important role in the pathogenesis of GPP.

Objective To examine the levels of fractional exhaled nitric oxide in bronchial asthma and the cor-relation between fractional exhaled nitric oxide and pulmonary function tests as well as the value of pulmonary function tests in curing the children with bronchial asthma. Methods Seventy - two asthmatic children came from Guiyang Ma-ternal and Child Health Hospital were studied. They were divided into 2 groups:mild - to - moderate asthma group(36 cases)and serious asthma group(36 cases). Fractional exhaled nitric oxide level and the lung function were measured and the correlation analysis was analyzed by SPSS 13. 0 software. Results The fractional exhaled nitric oxide concen-tration of the serious asthma group was significantly higher than that of the mild - to - moderate asthma group[(47 ± 26)ppb vs(29 ± 15)ppb,t = 3. 543,P ﹤ 0. 01],1 - second rate,maximum mid - expiratory flow(MMEF),and peak expiratory flow(PEF)in mild - to - moderate asthma group were significantly higher than those in serious asthma group[(99. 74 ± 9. 81)% vs(87. 17 ± 13. 30)% ,(87. 12 ± 35. 61)% vs(68. 90 ± 30. 88)% ,(99. 97 ± 10. 83)% vs (94. 31 ± 17. 69)% ](P ﹤ 0. 01,0. 05,0. 05). No correlations were found between fractional exhaled nitric oxide level and the lung function parameters,including forced expiratory volume in 1s(FEV1),1 - second rate,MMEF and PEF in the mild - to - moderate asthma group(r = 0. 073,0. 087,0. 242,0. 326,all P ﹥ 0. 05). However the level of fractional exhaled nitric oxide was negatively correlated with the lung function parameters,including FEV1,FEV1 / FVC,MMEF and PEF in serious asthma group(r = - 0. 354,- 0. 573,- 0. 478,- 0. 471,P ﹤ 0. 05,0. 001,0. 005,0. 005),respec-tively. Conclusions Monitoring of fractional exhaled nitric oxide and the lung function parameters can effectively indi-cate the state of illness of asthmatic child,which plays a key role in the treatment.

Objective To explore the application value of fiberoptic bronchoscopy in the diagnosis and treatment for refractoriness pneumonia in children.Methods Sixty-two cases of refractoriness pneumonia who had not been cured with normal anti-infective therapy for 2 weeks in Guiyang Children′s Hospital from Jun.2005 to Jun.2009 were selected.Fiberopic bronchoscopy Olympus BF-3C30 or BF-P40 examination was applied after routine local infiltration anesthesia to diagnose and local treat refractoriness pneumonia in 62 children.Results There were 27 cases(43%) of simple inflammation,20 cases(32%) of foreign bodies with concurrent infection,7 cases(11%) of dndobronchial tuberculosis,4 cases(6%) of dysplasia with concurrent infection,2 cases(3%) of bronchiectasis,and 2 cases(3%) of pulmonary hemosiderosis,among them,dysplasia were diagnosed on children under 1 year old,foreign bodies with infection on 1-3 years old,and tuberculosis and bronchiectasis on school age.Eleven cases(40%) of simple inflammation were identified various pathogen by bronchoalveolar la-vage fluid culture,mycoplasma PCR test and bronchial brushing and microscopic examination.Forty-seven cases of bronchi obstruction and stenosis caused by inflammation were irrigated and treated by drug and achieved satisfactory,among them,15 cases(79%) of pulmonary atelectasis were recovered with the treatment of fiberoptic bronchoscopy within 2 weeks,14 cases(70%) of foreign bodies were succeed in taking choked thing from bronchus with fiberoptic bronchoscopy.Conclusion The fiberoptic bronchoscopy play an important role in the etiological diagnosis and therapy of refractoriness pneumonia in children.

Objective: To know the pre-treatment drug resistance ( PDR ) status of newly reported human immunodeficiency virus type 1 ( HIV-1 ) infected individuals in Wenzhou, so as to provide guidance for antiretroviral therapy ( ART ). Methods: Totally 232 plasma samples of newly reported HIV-1 infected individuals who had not received ART were collected in Wenzhou in 2019. Virus ( HIV-1 ) RNA was extracted, followed by reverse transcription PCR and nested PCR to amplify the pol region and sequence. Resistance mutations and resistance to non-nucleoside reverse transcriptase inhibitors ( NNRTIs ), nucleoside reverse transcriptase inhibitors ( NRTIs ) and protease inhibitors ( PIs ) was analyzed. Results: The pol region sequences from 199 infected patients were obtained and the incidence of PDR was 8.04% ( 16/199 ). Eight genotypes were detected, including circulating recombinant forms ( CRFs ) CRF07_BC ( 47.24%, 94/199 ) and CRF01_AE ( 29.15%, 58/199 ) which were the dominant types. Two unique recombinant forms ( URFs ) were detected, namely URF( CRF01_AE/BC ) and URF( B/C ) . Thirty-one cases ( 15.58% 31/199 ) had drug-resistant mutations. For NNRTIs, NRTIs and PIs, 20 cases ( 64.52% ) , 2 cases ( 6.45% ) and 9 cases ( 29.03% ) with drug resistance mutations were detected, respectively. The resistance mutations to NNRTIs included K101E, K103N/R, V106I, E138K, V179D/E/T, Y181C, G190A and H221Y. Four cases each had two resistance mutations to NNRTIs. The resistance mutations to NRTIs were V75M and M184V. The resistance mutations to PIs were M46I, L33F and Q58E. For the newly released NNRTI drug Doravirine ( DOR ), two cases were found to have mutations of resistance. Conclusions The incidence of PDR among newly reported HIV-1 patients in Wenzhou is 8.04%, mainly caused by NNRTIs drug-resistant mutation. Resistance to the new drug DOR has emerged. The surveillance of drug resistance should continue to be strengthened.

Most chemical medicines have polymorphs. The difference of medicine polymorphs in physicochemical properties directly affects the stability, efficacy, and safety of solid medicine products. Polymorphs is incomparably important to pharmaceutical chemistry, manufacturing, and control. Meantime polymorphs is a key factor for the quality of high-end drug and formulations. Polymorph prediction technology can effectively guide screening of trial experiments, and reduce the risk of missing stable crystal form in the traditional experiment. Polymorph prediction technology was firstly based on theoretical calculations such as quantum mechanics and computational chemistry, and then was developed by the key technology of machine learning using the artificial intelligence. Nowadays, the popular trend is to combine the advantages of theoretical calculation and machine learning to jointly predict crystal structure. Recently, predicting medicine polymorphs has still been a challenging problem. It is expected to learn from and integrate existing technologies to predict medicine polymorphs more accurately and efficiently.

OBJECTIVETo observe MET-associated alteration during the trans-differentiation from MSCs to neuron-like cells, and to explore the possible molecular mechanism.

METHODSBone marrow MSCs were isolated from rat femur and purified in continuous cell culture. After induced differentiation to neuron-like cells by the combination of butylated hydroxyanisole (BHA) and dimethyl sulfoxide (DMSO), cells were tested by comparative polymerase chain reaction (PCR) for the relative expression of MET biomarkers and transcription factors, and for cell cycle by flow cytometry. Meanwhile, target genes of Wnt/β-catenin pathway were also analyzed by comparative PCR to determine the possible involvement.

RESULTSIn MSC-induced neuron-like cells, MET-associated transcription factors such as Snail, Slug, ZEB1, ZEB2, and Twist were significantly attenuated in expression level. The Mesenchymal marker Vimentin expression level was increased. Membrane protein E-cad was slightly down-regulated, while N-cad level was marginally elevated. Percentage of proliferating cells (S phase in cell cycle) markedly shrank from 40.42% for MSCs to 6.76% for MSC-derived neuron. Additionally, Wnt/β-catenin target genes β-catenin and c-myc were decreasingly expressed.

CONCLUSIONChemically induced trans-differentiation from MSC to neuron caused similar MET-featured alteration in gene expression and proliferation to known MET, which might be underlied by deactivation of Wnt/β-catenin pathway.

Animals ; Epithelial-Mesenchymal Transition ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism ; Rats ; Wnt Signaling Pathway ; beta Catenin ; metabolism

Objective To propose a new suggestion for the clinical downstaging of nasopharyngeal carcinoma (NPC) in the era of intensity-modulated radiotherapy (IMRT) without changing the current T,N,and M staging system.Methods We reviewed the records of 536 NPC patients treated in Sun Yat-Sen University Cancer Center from January 2002 to December 2006.The Kaplan-Meier method was used to calculate the disease-specific survival (DSS) rate,and the log-rank test was used for survival difference analysis.The Cox regression model was used to calculate the hazard ratio (HR) of each subset.ResultsAccording to the 7th edition of UICC/AJCC staging system,the 5-year DSS rates of stage Ⅰ-Ⅲ patients (except T3N2M0) were all more than 85%(P>0.05),those of stage ⅣA and ⅣB patients were 71.8% and 46.2%,respectively (P=0.171),and that of stage ⅠVC patients was only 24.0%.In stage Ⅲ,the 5-year DSS rate of non-T3N2M0 patients (91.5%) was significantly higher than that of T3N2M0 patients (78.6%)(P=0.042),but there was no significant difference in DSS between T3N2M0 patients and stage ⅣA and ⅣB patients.Based on the above results,new stage Ⅰ included T1-3N0-1M0 and T1-2N2M0,new stage Ⅱ included T3N2M0,T4N0-2M0,and TxN3M0,and new stage Ⅲ included TxNxM1.The 5-year DSS rates of new stage Ⅰ,Ⅱ,and Ⅲ patients were 93.3%,72.7%,and 24.0%,respectively (P=0.000).Compared with new stage Ⅰ patients,new stage Ⅱ and Ⅲ patients had HRs of 4.01 and 16.76,respectively,for 5-year DSS.Conclusions In the era of IMRT,the new clinical staging system (stages Ⅰ,Ⅱ,and Ⅲ) helps with prognostic evaluation and clinical treatment.