The paper reported two methods. a genetic probe and the solid phase radio-immunological assay, for the detection of the thermolabile toxin produced by Escherichia coli. The former is the application of a 32P-IabelIed probe of B subunit DNA prepared from Hind Ⅲ restriction fragment of LT-encoding DNA. and the latter is by using CNBr-activated paper as carrier and 125I-SPA to substitute the secondary antibody. Applying the methods to 160 strains of E. coli isolated from patients suffering from infantile diarrhea, the results showed that 42 out of 160 strains belong to the strain which produces the thormolabile toxin. These two methods not only have the advantages of being sensitive and specific, but also are beneficial favourable to the epidemiological survey and clinical diagnosis, because the testing bacteria are decomposed on the nitrocellulose membrane, so that hundreds of samples can be examined in one test.

Objective To investigate the action mechanism of blood-stage treatment that affects psoriasis vulgaris using observation of the Th17/Treg expression.Methods A total of 32 patients ( blood heat group,n =17 ; blood stasis group,n =15) and 15 healthy people ( control group,n =15 ) were observed.The frequencies of Th17 and Treg in peripheral blood were detected by flow cytometric analysis in patients before and after treated by heat-clearing and blood-cooling decoction,qi-enriching and blood-activating decoction.Results The ratio of Th17/Treg in peripheral blood [ blood heat group (4.21 ± 0.52 )％ ;blood stasis group( 3.32 ± 0.43 )％ ] was significantly increased in patients compared with controls [ ( 1.79 ±0.18)％ ] ( P ＜0.01 ).After herbal treatment,the ratio of Th17/Treg expression in the blood heat group [ ( 2.41 ± 0.22 ) ％ ] and in blood stasis group [ ( 2.02 ± 0.12 ) ％ ] was significantly decreased ( P ＜ 0.05 ),respectively.Conclusions Blood-stage treatment works well on Th17/Treg expression in patients with psoriasis vulgaris.

Objective To investigate the effects of RNA interference (RNAi) on Caspase-3 expression.Methods According to the gene sequence and the secondary structure characteristics of Caspase-3, three siRNA （Si-Caspase-3-1, Si-Caspase-3-2 and Si-Caspase-3-3） were designed with 21 basic group, and siRNAs against Caspase-3 gene were synthesized. With the liposome conduct, the synthesized siRNAs were transferred into neuroglia cells. After the cells crept the slices, cells were dyed with fluor Hoechst33258; the apoptotic cells were stained with TUNEL, the normal neuroglia cells were as control, the Caspase-3 protease activity were detected with hemi-quantitative RT-PCR and Western blot.Results The synthesized siRNAs could inhibit the expression of Caspase-3 gene. For Si-Caspase-3-3, the expression inhibition rate of Caspase-3 mRNA was 86.32% in neuroglia cell.Conclusion The synthesized siRNAs can inhibit the expression of Caspase-3 gene at cell level.

Objective To study the reasons of seroma formation and long-term existence after modified radi-cal mastectomy ( MRM) .Methods The structure of fibrous lamina for seroma postoperative breast cancer was ana-lyzed by pathological methods;the composition of drainage fluids after MRM in the different postoperative time peri-ods was determined;and the structure of fibrous lamina following an experimental mastectomy model created in rats was investigated,which the process of fibrous lamina formation was imitated.Results There were three layer struc-tures in fibrous lamina,including fibrous leakage,capillaries and fiber lay.Hematology test results supported seroma for exudate.Animal model showed that with the extension of time,fibrous lamina thickened gradually.Conclusion It is capillaries in fibrous lamina that seroma long-standing histological basis.Maybe excising or destructing the structure of subcutaneous fibrous lamina,making the procedure of wound healing to begin again as soon as possible,which is a kind of effective method to solve the problem of seroma long-term existence after MRM.

Objective To understand non stomatology undergraduates'strategies for learning stomatology and to study the reform on this course.Methods The learning strategies of 560 undergraduates majoring in clinical medicine from grade 2007 were investigated s and ten related factors like learning attitudes were investigated by learning strategies scale.Correlation analysis and linear regression analysis were applied to deal with research data.Results Most undergraduates were lack of strategies in learning stomatology.Related coefficient between 10 factors and academic scores ranged from 0.197 to 0.401,existing positive correlation(P＜0.05).Determination coefficients(R2)of attitude,motivation,time management and learning auxiliary means were 0.146,0.167,0.223and 0.122 respectively,which can be used to predict the scores of examination.Condusions Non stomatology undergraduates'strategies for learning stomatology is a vital factor influencing their academic scores.It's necessary for teachers to improve their teaching methods considering students'professional characteristics and learning strategies.

OBJECTIVETo establish subseries cell lines from single, cancer cell of Tca8113M1 cell line and detect the cancer stem cell markers in the different subseries cell lines.

METHODSThe subseries cell lines from single cancer cell of Tca8113M1 cell line were established by limiting dilution assay in vitro. The characteristic of tumorigenicity and CD44, CD184, extracellular soluble antigen (ESA) of the cancer stem cell markers were detected by xenotransplantation and flow cytometry respectively.

RESULTSTotal 192 single cells of Tca8113M1 cell line were cultured and were deposited as one cell per well. There were 12 subpopulations origin from 192 single cells spheroid cultures. The ratio was 6.25% (12/192). In the different subpopulations, the tumorigenicity and expression of CD44 and ESA were at high levels, but the expression of CD184 was in different level. There were three kinds morphology of colonies derived from single cancer cells, holoclone, meroclone and paraclone. Cell line could be derived from carcinoma cell holoclones by cell culture. Meroclone and paraclone did not exist in cell culture in vitro.

CONCLUSIONTongue cancer stem cell may exist in Tca8113M1 cell line, cell line can be established and holoclone is the origin of cell line. This is a novel approach to the identification and enrichment for cancer stem cell.

Objective:To evaluate the clinical efficacy and safety of a skin care ointment containing oligomeric maltose X in the adjuvant treatment of eczema-related pruritus.Methods:A multicenter, randomized, double-blind, vehicle-controlled clinical study was conducted. From March to September 2021, outpatients with mild to moderate eczema were collected from departments of dermatology of 4 hospitals, including Beijing Friendship Hospital, Hebei Traditional Chinese Medical Hospital, the Third People′s Hospital of Hubei Province, and Taizhou Central Hospital in Zhejiang Province. The patients were randomly divided into two groups by using a random number table: observation group topically treated with a skin care ointment containing oligomeric maltose X, and vehicle control group topically treated with an ointment vehicle. The ointments were applied during the attacks of itching for 14 consecutive days. Visits were scheduled before, 7, and 14 days after the start of the adjuvant treatment. The efficacy was evaluated according to the eczema area and severity index (EASI) and visual analog scale (VAS) , and adverse events were recorded. The efficacy and safety analyses were conducted by using chi-square test and t test. Results:Totally, 232 patients with eczema were enrolled, including 90 males and 142 females, with the age being 40.13 ± 13.36 years; 156 patients were in the observation group, and 76 in the vehicle control group. Before the adjuvant treatment, there were no significant differences in EASI (2.07 ± 2.24 points vs. 2.29 ± 2.28 points) or VAS (6.22 ± 1.78 points vs. 6.20 ± 1.79 points) scores between the observation group and vehicle control group ( t = -0.70, 0.06, P = 0.486, 0.955, respectively) . After one-day treatment, the VAS scores significantly decreased compared with the baseline scores in the two groups ( P < 0.001, P = 0.003, respectively) . After 14-day treatment, the VAS score was significantly lower in the observation group (2.67 points) than in the vehicle control group (3.35 points; t = -2.28, P = 0.024) . After 7- and 14-day treatment, the EASI scores significantly decreased compared with the baseline scores in both the two groups (all P < 0.001) , but there were no significant differences between the two groups ( P = 0.853, 0.731) . No adjuvant treatment-related adverse events were recorded in either of the two groups. Conclusion:The skin care ointment containing oligomeric maltose X is safe and effective in the adjuvant treatment of eczema-related pruritus, and can be applied to clinical practice.

Objective To explore the mechanism of tumor-associated fibroblasts(CAFs)for regulating proliferation and migration of prostate cancer(PCa)cells.Methods We conducted a bioinformatics analysis to identify miRNAs with high expression in PCa.The proliferation,migration and hsa-miR-18b-5p expression levels were observed in PCa cells co-cultured with CAFs.We further examined hsa-miR-18b-5p expression level in 20 pairs of PCa and adjacent tissue samples and in different PCa cell lines and normal epithelial cells using RT-qPCR.In PCa cell lines C4-2 and LNCAPNC,the effects of transfection with a hsa-miR-18b-5p inhibitor on cell proliferation,migration,invasion,drug resistance,apoptosis and cell cycle were evaluated,and the effects of has-miR-18b-5p knockdown on C4-2 cell xenograft growth and mouse survival were observed in nude mice.Dual luciferase reporter gene assay was used to validate the targeting relationship between hsa-miR-18b-5p and its target genes,whose expressions were detected in PCa cells using RT-qPCR and Western blotting.Results The expression of hsa-miR-18b-5p was significantly increased in the co-culture of CAFs and PCa cell lines,which exhibited significantly enhanced proliferation and migration abilities.Transfection with has-miR-18b-5p inhibitor strongly attenuated the effect of CAFs for promoting proliferation and migration of PCa cells,and in C4-2 and LNCAP cells cultured alone,inhibition of hsa-miR-18b-5p obviously suppressed cell proliferation,migration,invasion,and drug resistance.In the tumor-bearing mice,hsa-miR-18b-5p knockdown in the transplanted cells significantly inhibited xenograft growth and increased the survival time of the mice.Target gene prediction suggested that FBXL3 was a potential target of hsa-miR-18b-5p,and dual luciferase reporter gene confirmed a binding site between them.In C4-2 and LNCAP cells,hsa-miR-18b-5p knockdown resulted in significantly increased expression levels of FBXL3.Conclusion CAFs promotes proliferation and migration of PCa cells by up-regulating hsa-miR-18b-5p to suppress FBXL3 expression.

Objective To explore the mechanism of tumor-associated fibroblasts(CAFs)for regulating proliferation and migration of prostate cancer(PCa)cells.Methods We conducted a bioinformatics analysis to identify miRNAs with high expression in PCa.The proliferation,migration and hsa-miR-18b-5p expression levels were observed in PCa cells co-cultured with CAFs.We further examined hsa-miR-18b-5p expression level in 20 pairs of PCa and adjacent tissue samples and in different PCa cell lines and normal epithelial cells using RT-qPCR.In PCa cell lines C4-2 and LNCAPNC,the effects of transfection with a hsa-miR-18b-5p inhibitor on cell proliferation,migration,invasion,drug resistance,apoptosis and cell cycle were evaluated,and the effects of has-miR-18b-5p knockdown on C4-2 cell xenograft growth and mouse survival were observed in nude mice.Dual luciferase reporter gene assay was used to validate the targeting relationship between hsa-miR-18b-5p and its target genes,whose expressions were detected in PCa cells using RT-qPCR and Western blotting.Results The expression of hsa-miR-18b-5p was significantly increased in the co-culture of CAFs and PCa cell lines,which exhibited significantly enhanced proliferation and migration abilities.Transfection with has-miR-18b-5p inhibitor strongly attenuated the effect of CAFs for promoting proliferation and migration of PCa cells,and in C4-2 and LNCAP cells cultured alone,inhibition of hsa-miR-18b-5p obviously suppressed cell proliferation,migration,invasion,and drug resistance.In the tumor-bearing mice,hsa-miR-18b-5p knockdown in the transplanted cells significantly inhibited xenograft growth and increased the survival time of the mice.Target gene prediction suggested that FBXL3 was a potential target of hsa-miR-18b-5p,and dual luciferase reporter gene confirmed a binding site between them.In C4-2 and LNCAP cells,hsa-miR-18b-5p knockdown resulted in significantly increased expression levels of FBXL3.Conclusion CAFs promotes proliferation and migration of PCa cells by up-regulating hsa-miR-18b-5p to suppress FBXL3 expression.

ObjectiveTo demonstrate the long-term effect of Yishi Therapy for primary tinnitus and analyze the factors affecting the effectiveness. MethodsWe conducted long-term follow-ups of primary tinnitus cases treated with Yishi Therapy during 1 January 2017 to 30 June 2023 by face-to-face， online and telephone between 1 July 2023 and 31 January 2024， to understand the final regression of primary tinnitus and lifestyle changes. We retrospectively analysed the basic data of patients before treatment， and statistically analysed the recovery rate and overall effective rate of tinnitus treated by the Yishi Therapy， and the factors affecting the effectiveness such as lifestyle， age， duration of disease， gender， tinnitus location， severity of tinnitus， accompanying symptoms and complications on the overall effective rate. ResultsA total of 496 cases of primary tinnitus completed follow-ups successfully， and the total Tinnitus Evaluation Questionnaire （TEQ） at follow-up was significantly lower than that before treatment （P＜0.001）. Among the 496 cases， 101 （20.36%） cases cured， 189 （38.10%） cases showed significant improvement， 116 （23.39%） cases were effective， 90 （18.15%） cases were ineffective， and the overall effect rate was 81.85% （406/496）. At follow-up， the score of lifestyle was higher than that before treatment （P＜0.001）. Grouped by the score of lifestyle at follow-up， the effective rates of better lifestyle group and poor lifestyle group were 97.53% （237/243） and 66.80% （169/253）， respectively， with statistical significance （P＜0.001）. Grouped by the mean value of the difference in lifestyle scores before and after treatment， the effective rates of the group with greater lifestyle adjustment and the group with smaller adjustment were 97.55% （239/245） and 66.53% （167/251）， respectively， and the differences were statistically significant （P＜0.001）. Binary logistic regression showed that the better lifestyle， the greater lifestyle adjustment and the severity of tinnitus were positively correlated with the total effective rate of tinnitus （P＜0.001）， while older and hearing loss were negatively correlated with the total effective rate of tinnitus （P＜0.05）. ConclusionYishi Therapy on primary tinnitus shows good long-term effect， lifestyle， severity of tinnitus， hearing loss or not， age are the relevant factors affecting the efficacy of tinnitus， lifestyle is the main factor affecting clinical efficacy.