Objective To optimize the extraction process of free anthraquinones from Rheum Emodi Wall. with microwave technique. Methods Single factor and orthogonal test design were used to investigante the effects of the four main factors including microwave power, alcohol concentration, extraction time and the ratio of materials and solution on the extraction efficiency of free anthraquinones. Results Using 70% ethanol solution in high-fire and the ratio of materials and solution of 1∶8 for 5 min to extract two times, the extraction rate of Rhubarb anthraquinones was 1.31% and the relative rate was 88.52%. Conclusion Microwave extraction method is high efficient, easy and timesaving for extracting anthraquinones in Rheum Emodi Wall..

Objective:To isolate breast cancer stem/progenitor cells from human breast cancer and study their proliferation and differentiation biological characteristics over long-term passages in vitro. Methods:Human breast cancer stem/progenitor cells were enriched in suspension cultures as nonadherent mammospheres(MS). Serial sphere formation assay was performed to determine self-renewal ability of mammosphere-derived cells (MSDC). Differentiation was induced by culturing MSDC in DMEM-F12 supplemented with serum but without growth factors. The ratio of CD44~+/CD24~(-/low) cell population was evaluated by flow cytometry(FCM). Results:The mammospheres were formed after inoculation of primary breast cancer cells into culcutre medium with growth factors but without serum. The mammospheres contained undifferentiated cells similar to stem cells, which had self-renewal and extensive proliferation capabilities. With increasing passages, the cells tended to adhere and differentiate. The number of adhering and differentiating cells increased, and the amount and size of mammospheres decreased. The CD44~+/CD24~(-/low) cell population was enriched in the basal-like molecular subtype of human breast tumors. The biological behaviors of mammospheres varied between different specimens.Conclusion:Cancer cells with stem cell properties of self-renewal, indefinite proliferation and multi-lineage differentiation widely existed in human breast cancer tissues. The biological behaviors varied because of different origin of specimens and changed under the effects of environmental factors.

Objective: To investigate a cluster of coronavirus disease 2019(COVID-19)caused by latent infection in Haikou,so as to provide reference for the prevention and control of COVID-19 clusters. Methods: An epidemiological investigation was conducted according to the COVID-19 Prevention and Control Program(Fourth Edition). The course of diagnosis and treatment,clinical characteristics and field investigation data were collected to analyze the transmission chain and the intergeneration between cases. Results : Among 39 people involved,five confirmed cases and two asymptomatic infections were found,with an attack rate of 17.95%. The cases aged from 40 to 65 years and lived in the dormitories near the farm of Dongshan. The first case(named Case 1)closely contacted with a confirmed case of COVID-19 in Chengmai from January 28 to February 10,was isolated on February 13 and developed symptoms on February 16. The other six cases(Case 2 to Case 7)shared the water source with Case 1 from January 28 to February 13(within the incubation period of Case 1). They needed to open the power switch outside Case 1's room and the water valve in Case 1's washroom before the use of water. They might be infected by contacting the doorknob and the water valve contacted by Case 1,or by the aerosol formed after Case 1 used the washroom,then infected each other when living together. The onset of Case 2 to Case 7 was earlier than Case 1,and they had no travelling history in Hubei Province 14 days before and contacted no confirmed cases except Case 1. Therefore,Case 1 was the source of the cluster during his incubation period of COVID-19. Conclusion This was a cluster of COVID-19 due to latent transmission by living in the same area and touching the same objects indirectly,which indicated that COVID-19 was infectious in the incubation period.

OBJECTIVETo investigate the effect of 1,25-dihydroxyvitamin D(3) and 5-fluorouracil, either alone or in combination, on the expression of IGFBP-3 in human esophageal carcinoma 109 cell xenograft in nude mice.

METHODSIn vitro cultured esophageal carcinoma Eca-109 cells were inoculated subcutaneously in BALB/c mice. The tumor-bearing mice were randomly divided into control group (A), 1,25-dihydroxyvitamin D(3) group (B), 5-fluorouracil group (C), and 1,25-dihydroxyvitamin D(3) plus 5-fluorouracil group (D). 1,25-dihydroxyvitamin D(3) and 5-fluorouracil were administered at the doses of 2.5 ug/kg and 25 mg/kg via intraperitoneal injections, respectively, and the mice in the control group received saline injection only. The tumor growth was observed and the expression of IGFBP-3 in the tumor xenograft was detected using immunohistochemistry. An automatic biochemistry analyzer was used to determine serum calcium levels, and Von Kossa staining was utilized for observation of calcium deposition in the kidneys.

RESULTSCompared with that in group A, the xenograft in groups B, C, and D all showed a lowered growth rate with a smaller tumor volume, and presented with stronger IGFBP-3 positivity and significantly higher levels of IGFBP-3 protein expression (P<0.05). In group D, the protein expression of IGFBP-3 was significantly increased compared with that in groups B and C (P<0.05). Compared with that in group A, serum calcium level was slightly increased in groups B, C, and D, , but no obvious calcium deposition was found in the kidney tissue sections.

CONCLUSIONBoth 1,25-dihydroxyvitamin D(3) and 5-fluorouracil can inhibit the growth of the tumor xenograft in nude mice, and their combination is more effective. This effect is probably associated with increased protein expression of IGFBP-3 in the xenograft tumor. No calcium deposition occurs in the kidney tissue of the tumor-bearing mice.

Animals ; Cell Line, Tumor ; Fluorouracil ; pharmacology ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Vitamin D ; analogs & derivatives ; pharmacology ; Xenograft Model Antitumor Assays

Objective To understand the prevalence of autism spectrum disorders (ASD) in children aged 0-6 years old and influencing factors in Hainan province.Methods A total of 37 862 children aged 0-6 years were selected from 18 counties in Hainan province for a screening by using questionnaire of "warning signs in child development",then field diagnosis was made,and general descriptive statistic analysis was conducted.The prevalence of ASD and related factors were analyzed with x2 test and unconditional logistic regression model.Results Among 37 862 children aged 0-6 years,235 were diagnosed with ASD,the prevalence of ASD was 0.62％ (0.99％ in boys,0.17％ in girls),the differences was significant (x2=101.91,P=0.000).The prevalence of ASD increased with age (x2=288.62,P=0.000).The prevalence of ASD was significantly higher in urban area than in other areas (x2=114.77,P=0.000).Factors such as full term pregnancy or not,neonatal asphyxia,father' s characteristics,father' s habit of chewing areca or smoking,mother' s general mood,and mother' s induced abortion history were the influencing factors for ASD.Conclusion The prevalence of ASD in children aged 0-6 years was high in Hainan and was influenced by genetic factors,pregnancy and delivery process,parents unhealthy habit before and during pregnancy and other factors.

Objective To investigate the genetic carrier rate of thalassemia and its gene mutation types as well as the distribution characteristics among couples of reproductive age in Dongfang City of Hainan Province,and to provide a basis for making prevention and control strategies against thalassemia.Methods Samples were collected from 1 000 couples undergoing premarital and pregestational screenings for thalassemia in Dongfang City of Hainan Province from September 2012 to March 2013,in which the positive ones in preliminary screening were further tested by genetic diagnoses and the genotypes were retrospectively analyzed.Results Among 1 000 couples,322 spouses were diagnosed with thalassemia gene mutation and the carrying rate was 16.10％ (322/2 000).In those carriers,246 spouses were α-thalassemia and the carrying rate was 12.30％ (246/2 000),accounting for 76.40％(246/322) of all thalassemia carriers,among them,there were 197 cases of α-deficiency genotype,accounting for 61.18％ (197/322),32 carried mutated α-gene,accounting for 9.94％ (32/322),17 carried both deleted and mutated α-gene,accounting for 5.28％ (17/322);43 spouse were β-thalassemia and the carrying rate was 2.15％(43/2 000);33 spouse were both α-and β-thalassemia and the carrying rate was 1.65％ (33/2 000).In spouses diagnosed with α-thalassemia,the major genotype was-α37/αα,accounting for 19.25％ (62/322);the second ranked was-α4.2/αα,accounting for 17.70％ (57/322),and the third ranked was--SEA/αα,accounting for 8.70％ (28/322).In spouses diagnosed with β-thalassemia,the major genotype was CD41-42/N,accounting for 9.63％ (31/322).Conclusions The population carrying rate of thalassemia in Dongfang City of Hainan Province is high,and its major type is α-thalassemia.For the purpose of decreasing the birth rate of thalassemia,major,local public health department should attach great importance to thalassemia prevention,and strengthen premarital and pregestational screening for thalassemia.