Objective To explore the inhibition effects of epidermal growth factor receptor (EGFR) antagonist monoclonal antibody cetuximab (C225) on breast cancer stem cells in human breast cancer cell line MCF-7.Methods The effects of C225 on the proliferation of breast cancer cell line MCF-7 were detected by MTT assay. MCF-7 cells were cultured to generate primary mammospheres, and were divided into control group, C225 group, epidermal growth factor (EGF) group and EGF + C225 group according to whether or not the culture media contained exogenous EGF and C225.Thirteen days after culture, the volume and number of mammospheres of these four groups were observed, and mammosphere-forming efficiency ( MFE) was calculated. The percentages of CD44~+ CD24~- cells in mammospheres of these four groups and in routinely cultured MCF-7 cells were determined by flow cytometry. Results The inhibition rate on MCF-7 cells increased with the concentration of C225.Compared with control group, the volume of mammospheres in C225 group significantly decreased, and MFE and percentages of CD44~+ CD24~- cells in mammospheres significantly decreased [(0.61 ±0.04)% vs (1.44±0.09)%, P<0.01; (3.50±0.29)% vs (9.07 ±0.52) % , P<0.01]. Compared with EGF group, the volume of mammospheres in EGF + C225 group significantly decreased, and MFE and percentages of CD44~+ CD24~- cells in mammospheres significantly decreased [ (0.68 ± 0.04) % vs (1.61 ± 0.05) % , P < 0.01; (4.00 ± 0.58) % vs (10.47 ± 0.79) % , P < 0.01]. The percentage of CD44~+ CD24~- cells in routinely cultured MCF-7 cells was (2.03 ±0.15) % , and was significantly different from those in EGF group and control group (P < 0.01). There was no significant difference in volume of mammospheres, MFE and percentage of CD44~+ CD24~- cells in mammospheres between EGF group and control group (P >0.05). Conclusion C225 has significant inhibition effects on CD44~+ CD24~- cells in MCF cells.

Objectives To study the effect of VIM in Enterovirus 71 (EV71) infection of (human brain microvascular endothelial cells (HBMEC) and elaborating the mechanism of EV71 infection in the nervous system. Methods Knocked out the VIM by CRISPR technology , the differences in EV71 absorption , replication , release between wild VIM and VIM knocked-out (VIM-KO) HBMEC were detected by fluorescence quantitative PCR. Results 4 ℃ absorption experiment conformed that EV71 adsorption in VIM- KO is 40% less than in the normal HBMEC. After EV71 infect HBMEC for 48 h (48 h p. i.), the quantitative PCR result showed intracellular viral RNA in VIM-KO was only 1/12 of that in the normal HBMEC. Also the extracellular viral RNA was quantified, and the number of cells in VIM-KO had been reduced 1.4 times compared with the normal HBMEC. Conclusions Once VIM knocking out, EV71 attachment has been obviously reduced. Meanwhile, the level of viral RNA replication and release are decreased compared with the normal HBMEC. VIM may be an attachment receptor of EV71 in HBMEC , when the virus invades HBMEC with the binding of VIM. Moreover , VIM plays an important role in the replication and release of EV71.

0.05). 1 h, 2 h , 3 h, and 4 h after co-culture, the inhibition rate in capsule-depleted group was higher than that of the wild strain group(P0.05). Conclusion: Capsule-depleted C. neoformans causes more significant endothelial cell injury than wild strain C. neoformans does, indicating that the capsule play an important role in C. neoformans-induced damage of endothelial cells.

A gram-negative bacillus was isolated from the bodies of dead clams among clambanks along the south beach of Jiangsu province. It was confirmed to the Vibrio furnissii by morphological and biochemical characteristic examinations. This isolate grows well in clam's body fluid media and may multiply rapidly in sea-water at temperature about 25—37℃. It is of high toxicity. The healthy clams were all diseased with the same syndrome and died as natural illness by inoculate them with the isolate. It is considered that the large enormous death of clams along the south beach of Jiansu province is concerned with the wide spreading of the clam's infectious disease caused by Vibrio furnissii which hasn't been reported both internal and abroad.

Objective To study the factors influencing epidemiological characteristics and virulence of Enterovirus 71 (EV71) in Guangdong province from 2008 to 2010. Methods RNA was extracted from collected samples or cultured virus , then reversing transcription into cDNA. We amplified full-length EV71-VP1 using poly-merase chain reaction technology , then conducted sequence alignment and established phylogenetic tree with MEGA software (version 5.0) to confirm the genotype of EV71. The association between severity of clinical symp-toms and sex, age, viral genotype and VP1 variation was also analyzed using Logistic regression. Results The genotype of the predominant epidemical strain was C4a in Guangdong from 2008 to 2010. However , this subtype had already differentiated into 4 subgroups (C4a1- C4a4). There was no correlation between clinical syndrome and sex or viral genotype; the severity of symptoms was negatively correlated with age: before 4 years old, varia-tion A289T can easily lead to severe cases, increasing the risk of infection (P<0.05, OR = 2.360, 95%CI:1.163~ 4.659). Conclusion The main epidemical EV71 strain is C4a1 in Guangdong province. The emerging differen-tiation and simultaneous prevalence should merit attention to strengthen relevant surveillance; and the protection of the susceptible population should be reinforced during EV71 prevalence.

The prosthesis fitting and the rehabilitation are discussed in this paper for 6 cases double above knee amputees hospitalized in the past six years. The author suggest that the prosthesis fitting and rehabilitation training are different between double and single above knee amputees. This paper expounds particularly the structure of temporary stubbie with peculiarity of rocker feet; the distinct demand of double knee joint of permanent prosthesis with peculiarity of artificial feet;the use of permanent stubbie in daily living. The important of physical strength training, the standing balance training, the walking functional training, some relative functional movement training and on problem of the amputees sitting wheelchair backward fall with preventive measures has been emphasized.Keyworks:amputation; prosthesis; rehabilitation

Objective To explore the detection trend of vaginal intraepithelial neoplasia(VaIN)of lower genital tract from 2013 to 2015. Methods A retrospective analysis was undertaken of colposcopy-directed biopsy of cervical, vaginal and vulvar intraepithelial neoplasia lesions include cervical intraepithelial neoplasia (CIN), VaIN and vulvar intraepithelial neoplasia (VIN) in Obstetrics and Gynecology Hospital of Fudan University from January 2013 to December 2015. Results (1) Overall data of CIN, VaIN and VIN:a total of 16732 cases were diagnosed of lower genital intraepithelial neoplasia in 3 years, accounting for 23.20% (16732/72128) of total colposcopy-directed biopsy cases. Among them, CIN, VaIN and VIN accounted for 19.48%(14053/72128), 2.67%(1923/72128), 1.05%(756/72128) of total colposcopy-directed biopsy cases of the lower genital tract, 83.99%(14053/16732), 11.49%(1923/16732), 4.52%(756/16732) of total lower genital intraepithelial neoplasia, respectively. (2) Annual data of CIN, VaIN and VIN from 2013 to 2015. The annual proportion of CIN in all intraepithelial neoplasia of lower gential tract was basically stable, consisting of 86.02%(3955/4598),83.25%(4795/5760) and 83.20%(5303/6374), respectively. The annual proportion of VaIN was gradually increasing, consisting of 8.09% (372/4598), 12.45%(717/5760) and 13.08%(834/6374), respectively. The annual proportion of VIN was gradually decreasing, consisting of 5.89% (271/4598), 4.31% (248/5760) and 3.72% (237/6374), respectively. Conclusion The increasing detection of VaIN from 2013 to 2015 might correlate with the increasing attention to inspection of the entire vaginal wall.

OBJECTIVE: To analyze the clinical, familial and hereditary features of myotonic dystrophy to improve the knowledge and provide molecule evidence for gene diagnosis and prenatal diagnosis of myotonic dystrophy or dystrophia myotonia (DM) families. METHODS: Clinical data of 2 DM families were collected based on the probands. The number of trinucleotide CTG repeat in the 3' untranslated region of myotonic dystrophy protein kinase (DMPK) gene on chromosome 19 was determined by DNA sequence and repeat fragment. RESULTS: Except for 1 subclinical patient, another 5 patients progressed slowly with the features of myotonic muscular weakness and atrophy. One patient had hatchet face, 1 had cataract and diabetes mellitus, and the other 3 were bald. Electromyologram showed 3 patients had myotonic discharge and myopathic abnormalities. The number of trinucleotide CTG repeat in the 3' untranslated region of DMPK gene of 5 patients exceeded 50. CONCLUSION DM can be anticipated. Gene analysis can verify the disease and identify subclinical patients. It helps to prevent the DM births by hereditary consultation performing prenatal diagnosis.
Adolescent ; Adult ; Female ; Humans ; Male ; Myotonic Dystrophy ; diagnosis ; genetics ; Myotonin-Protein Kinase ; Pedigree ; Polymerase Chain Reaction ; methods ; Protein-Serine-Threonine Kinases ; genetics ; Trinucleotide Repeats

Objective:To investigate the effect of bedside high-flow continuous blood purification (CBP) combined with Xuebijing in the treatment of severe sepsis (SS) and the influence on the patient′s coagulation-fibrinolysis index, immunity index and expression of peripheral blood Toll-like receptor 4 (TLR4).Methods:Ninety-three patients with SS who were admitted and treated in the Lianyungang First People′s Hospitalfrom January 2017 to October 2019 were selected. They were divided into the combined group (51 cases, treatment with bedside high-flow CBP and Xuebijing injection based on bundle therapy) and the control group (42 cases, treatment with Xuebijing injection based on bundle therapy). The changes in coagulation and fibrinolysis index, immunity index, biochemical index such as TLR4 before treatment and after 1 week of treatment were compared between the two groups. The incidences of complications in both groups were statistically analyzed, and the discharge time from ICU, mechanical ventilation time and 28-day mortality were recorded.Results:After 1 week of treatment, the levels of prothrombin time (PT) and activated partial thromboplastin time (APTT) in the two groups were shortened, D-dimer (D-D) and fibrinogen (FIB) were decreased ( P<0.05); and the levels of PT and APTT in the combined group were shorter than those in the control group, the levels of DD and FIB were lower than those in the control group, there were statistical differences ( P<0.05). After 1 week of treatment, the levels of CD 4+ and CD 4+/CD 8+ ratio in both groups were increased ( P<0.05), and the levels of CD 4+ and CD 4+/CD 8+ ratio in the combined group were higher than those in the control group ( P<0.05). After 1 week of treatment, the levels of TLR4, C-reactive protein (CRP), procalcitonin (PCT), white blood cell count (WBC), blood lactate (Lac), blood urea nitrogen (BUN) and serum creatinine (Scr) in both groups were decreased ( P<0.05), meanwhile, the above indexes in the combined group were lower than those in the control group ( P<0.05). The incidence of multiple organ failure and the 28-day mortality rate in the combined group were lower than those in the control group: 3.92%(2/51) vs. 19.05%(8/42), 13.73%(7/51) vs. 30.95%(13/42), there were statistical differences ( P<0.05). The discharge time from ICU and mechanical ventilation time in the combined group were shorter than those in the control group: (12.35 ± 2.14) d vs. (14.17 ± 3.36) d, (7.12 ± 2.23) d vs. (8.51 ± 2.39) d, there were statistical differences ( P<0.05). Conclusions:Bedside high-flow CBP combined with Xuebijing injection in the treatment of SS can improve the patient′s condition, regulate the balance of coagulation and fibrinolysis, avoide the activation of coagulation, inhibite inflammatory response, reduce the expression of TLR4 in peripheral blood, improve immune function, protecte kidney function and promotethe patient′s recovery.

This article was aimed to study the chemical constituents of Dioscorea opposita Thunb.The chemical constituents were isolated and purified by Diaion HP-20,Toyopearl HW-40,Sephadex LH-20,MCI Gel CHP-20,silica gel column chromatography and preparative HPLC,TLC,purification and isolation from Dioscorea opposita Thunb.The structures of isolated compounds were identified by thc physicochemical properties and spectral analysis.The result showed that 14 compounds were isolated from Dioscorea opposita Thunb.The chemical structures were elucidated as L-Tryptophane (1),Seguinosides F (2),1-methoxycarbonyl-β-carboline (3),Helichrysin A (4),Bungein A (5),Hydroquinone (6),Zarzissine (7),Cyclo-(Pro-Thr) (8),4-Hydroxy-3-methoxybenzyl alcohol (9),pyridine-3-carboxamide (10),Arbutin (11),Methyl syringate 4-O-β-D-glucopyranoside (12),L-Phenylalanine (13),1,2-benzenedicarboxylic acid,1,2-bis[2-(2-hydroxyethoxy) ethyl] ester (14).It was concluded that chemical compounds 1-14 were isolated for the first time from Dioscorea opposita Thunb.