Objective:To optimize the processing technology of processed Morinda Officinalis by taking monotropein content as the index. Methods:The amount of licorice,stir frying time and cooking pot temperature were used as the investigation factors in orthogonal experiments design,and an HPLC method was used to determine the content with he following chromatographic conditions:Kromalsil C18 chromatographic column(200 mm × 4. 6 mm,5μm),mobile phase of methanol-0. 4% phosphoric acid solution(90 ∶10),the column tem-perature at 30℃,the flow rate at 1 ml·min-1 ,the detection wavelength at 233 nm and the sample size of 5μl. Results:The monotropein content of each processed Morinda Officinalis sample after different processing was:0. 535 6, 0. 582 4, 0. 523 4, 0. 589 1, 0. 578 6, 0. 587 8,0. 575 2,0. 609 1 and 0. 558 7 mg·g-1,which showed that the processing technology could affect,the monotropein content. Conclusion:The best processing technology of preparation Morinda Officinalis based on Monotropein content is: Liquorice of 6%,stir time of 10min and boiling pot temperature at 100 ℃.

Objective Using mouse autoimmune premature ovary failure (POF) model to seek theoretical evidence for a possible clinical therapy of autoimmune POF with glucocorticoid (GC) or an-drogen. Methods After autoimmune POF was induced in 60 mice by Pzp3, the mice were randomly as-signed into 3 groups (n=20) : Two groups were treated with GC or androgen and the control group was treated with distilled water. We observed the changes in the sexual cycles of the mice, the serum level of AzpAb, infiltration of cells positively expressing CD45 in the ovary, and pathological alterations of the ovary. Results The sexual cycle of each therapy group was significantly different from that of the control group. The mean serum level of AzpAb of each therapy group was significantly lower than that of the con-trol group, and the mean serum level of AzpAb in the GC group was significantly higher than that of the androgen group. The percentage of growing follicles in the ovary of each therapy group was significantly higher than that of the control group. Ovaries infiltrated by cells positively expressing CD45 of each thera-py group were significantly fewer than those of the control group. Conclusion GC or androgen in mice with autoimmune POF could obviously ameliorate the pathogenetic conditions of the disorder, and both treatments have similar therapeutic efficacy.

Objective To detect PAH gene mutations in classical PKU patients by HRM analysis. MethodsMutation scanning of PAH gene were performed in 17 classical PKU patients by HRM analysis ( LightScanner), covering the 13 exons and exon-intron boundaries. The HRM results were further confirmed by DNA sequencing, and the sensitivity and specificity of HRM method in PKU diagnosis were also evaluated. In addition, prenatal diagnosis was performed in two fetuses at risk for classical PKU. Results In the 17 patients, two mutations were identified in 16 patients, three mutations were identified in 1 patient.In this subject, a total of 22 different pathogenic mutations : 194V( c. 280A ＞ G), IVS4nt-1 G ＞ A( c. 442-1G ＞ A), R158Q( c. 4736 ＞ A), Q160X( c. 478C ＞ T), W187X( c. 561G ＞ A), E6nt-96A ＞ G( c. 611A ＞G), G239D( c. 716G ＞ A), R241 C( c. 721C ＞ T), R243Q( c. 728G ＞ A), G247R (c. 739G ＞ C), G247V (c. 740G＞T), R261X(c. 781C ＞T), PR261Q(c. 782G ＞ A), H264R (c. 791A ＞ G), F302fsX39 (c. 904delT), E305K( c. 913G ＞ A), G312V( c. 935G ＞ T), Y356X( c. 1068C ＞ A ), V399V ( c. 1197A ＞T), R408Q(c. 1223G ＞ A), T418P(c. 1252A ＞ C) , A434D(c. 1301C ＞ A), 3 silent mutations Q232Q (c. 696G ＞ A), V245V(c. 735G ＞ A), L385L(c. 1155C ＞ G), and one single nucleotide polymorphism rs2280615 ( c. 402A ＞ C) were identified, of which 194V ( c. 280A ＞ G), Q160X ( c. 478C ＞ T), H264R (c. 791A ＞ G), G312V( c. 935G ＞ T) and E305K ( c. 913G ＞ A) were novel mutations identified in PAH gene. The prenatal diagnosis results of the two fetuses : one was diagnosed as normal, the other was diagnosed as a carrier. In this study, the sensitivity and specificity for mutation detection by HRM were 100％, and the HRM results were consistent with DNA sequencing results. Conclusions HRM analysis is a simple,accurate, rapid, high-throughput and low-cost genetic analysis approach. It could be applied to mutation scanning of classical PKU of PAH gene and rapid prenatal diagnosis in parents with known mutations.

Objective To identify the effects of androgen on mouse model of autoimmune ovary failure.MethodsFifty mice were randomly divided into five groups as control group,early therapeutic group and its model group,late therapeutic group and its model group.Autoimmune POF was induced in forty mice by Pzp3.Then,the animals were randomly divided into four groups: two were treated with androgen from the 15th and 20th day after immune.The control group was treated with distilled water.The therapeutic efficacy of each treatment was evaluated by the changes in sexual cycles of mice,the serum level of AzpAb,infiltration of cells positively expressing CD45 in the ovary as well as the percentage of growing follicles of the ovary.Results The sexual cycles,serum level of AzpAb,infiltration of CD45 cells in the ovary of each model group were more significant than those in control group;early therapeutic group was less than model group;and serum level of AzpAb in early therapeutic group was lower than that of late therapeutic group.The percentage of growing follicles in the ovary of each model group was lower than that of control group;early therapeutic group was higher than model group and late therapeutic group.Conclusion The use of androgen in mice with autoimmune POF may notably ameliorate the pathogenetic conditions of the disorder,and the treatment during prophase was apparently more effective than that during the advanced stage.

20?10 9/L) was observed (mean 41 days, range 12 to 118 days). There w as the association between the number of CD34 +CD44 + cells infusion and time to neutrophic recovery (?= -0.657 , P

Objective:To optimize the processing technology of prepared Morinda officinalis based on nystose content. Methods:Orthogonal experiments and the nystose determination method in Chinese Pharmacopoeia (2010 edition) were used to optimize the pro-cessing technology. A Welch AQ-C18(250 mm ×4.6 mm,5 μm)chromatographic column was applied with methanol-water(3∶97)as the mobile phase, the column temperature was at 25℃, the flow rate was 0. 8 ml·min-1 , the injection volume was 10μl, and the drift tube temperature was at 40℃ with low temperature atomization. Results:The linearity of nystose was good within the range of 0. 214 8-0. 644 4 mg·ml-1 and the recovery was 99. 5%(RSD=3. 5%,n=6). Conclusion: On the basis of nystose content determination, the optimal processing technology of prepared Morinda officinalis is as follows:liquorice of 4%, stir fry time of 15min and the boiling pot temperature of 200℃.

Objective To explore the molecular mechanisms of primary amenorrhea by using arrayCGH technology. Methods Ten patients with primary amenorrhea and 10 female volunteers with regular menstrual cycles as healthy controls were selected. All patients and control samples were analyzed by conventional chromosome analysis (G-banding technology) and array-CGH technology, respectively. ArrayCGH was performed using Affymetrix Cytogenetic 2. 7M arrays following the manufacturer's standard protocol. Results Both the patient group and control group analyzed by conventional G-banding karyotype technology showed a negative result with a normal female karyotype: 46, XX. The result of array-CGH analysis demonstrated a microdeletion of approximately 110 000 bp located at the end of the short arm of X chromosome [46, X, del (X) (p22. 33 )] were identified in 5 patients, which was not detected in the control group. All healthy control samples by array-CGH analysis showed no pathological DNA copy number variation. Conclusions Array-CGH technology can improve the diagnosis rate of chromosomal disease at the DNA level. It is necessary to provide array-CGH for higher resolution genetic analysis of idiopathic primary amenorrhea patient who can not be identified by conventional technology.

Objective To determine the value of spectral karyotyping(SKY)in identification of the marker chromosome.Methods Selected six cases that could not be identified in clinic were studied,using samples of peripheral blood from four cases,and samples of amonic fluid and fetal cord blood for prenatal diagnosis in two cases were investigated.All cases were analyzed with the routine SKY method.and the results with the SKY View software.The SKY results were identified by using fluorescence in situ hybridization(FISH).And C-banding technique was used to help diagnose the heterochromatin.Results SKY wag successfully performed on all of 6 cases.The origin of all marker chromosomes was identified by SKY.Except case No.4,the others were confirmed by FISH.It helped determine the pregnancy outcome in two cases of prenatal diagnosis:one case of genetic marker chromosome continued the pregnancy,and another case of de novo marker chromosome was terminated of the pregnancy.Conclusion SKY may be a vahable tool to diagnose the marker chromosome with rapidness,direct-viewing and sensitiveness.It can be used to assess the prognosis and the pregnancy outcome.

Objective To investigate the relationship between spontaneous miscarriage and embryonic chromosome abnormalities,and to evaluate the clinical application of karyotype analysis by chorionic villus cell culture. Methods The chorionic villus karyotype of 1983 cases of miscarriage from January 2010 to July 2016 in Guangzhou Women and Children′ s Mecical Center were analyzed retrospectively. The miscarried chorionic villi were obtained by curettage under sterilized condition. The chromosome specimens were prepared after chorionic villus cell culture. Karyotype analysis was performed by G-banding technique. Results In the 1983 samples, successful karyotype analysis was performed in 1770 cases, with the successful rate of 89.98%. Chromosomal abnormalities were found in 1038 cases (58.64%,1038/1770). Chromosomal structural abnormalities were found in 37 cases. The numeral abnormalities were more common than structural abnormalities, and most of the numeral abnormalities were aneupoidies. In turn, they were trisomy 16, 45,X, trisomy 22, trisomy 2, trisomy 21, trisomy 15. The most common structural abnormality was balanced translocation, including Robersonian translocation. Female embryoes accounted for 61.02%(1080/1770) miscarriages and for 57.4%(596/1770) of chromosomal abnormalities, while male embroyes acoounted for 61.02%(1080/1770),57.4%(596/1770)respectively. The proportion of female embryoes was higher than male embryoes. The median age of the patients was 30 years old(16-46 years old). As the maternal age increased, the proportion chromosomal abnormalities increased. The incidence of chromosomal abnormalities in the advanced age group (≥35 years) was 68.38%(240/351), which was significantly higher than that in the younger group (56.24% ,798/1419; χ2=17.10, P<0.01). Conclusions Embryonic chromosomal abnormalities are the most common cause of early spontaneous miscarriage. The abnormalities centralize in some karyotypes. There is certain relationship between maternal age and the incidence of miscarriage, as well as the embryonic gender. Chorionic villus cell culture and karyotype analysis are helpful in finding the cause of miscarriage and counsel the patients.

Objective To investigate the effects of D-tryptophan (D-Trp) on the formation of Streptococcus mutans (S. mutans) biofilm and the dispersal of 24 h-old biofilm, and the drug susceptibility of S. mutans against chlorhexidine (CHX) under the role of D-Trp. Methods Optical density assay was used to evaluate the growth curve of S. mutans exposed to 5.0 mmol/L D-Trp for 28 h. The non-treated group was not added with D-Trp. After treatment with 1.0, 2.5 and 5.0 mmol/L D-Trp, crystal violet staining was used to observe the changes of S. mutans biofilm formation in treatment group and non-treatment group. Crystal violet staining and confocal laser scanning microscopy (CLSM) were applied to illustrate the effects of 1.0, 2.5 and 5.0 mmol/L D-Trp on the dispersal of 24 h-old S. mutans biofilm. Resazurin sodium was used to indicate the effect of 5.0 mmol/L D- Trp on the minimum inhibitory concentration (MIC) and the minimum biofilm inhibitory concentration (MBIC) of treatment groups and negative control group. Results The growth curves of planktonic S. mutans within 28 h was consistent in treatment group and the non-treated group, both attained exponential phase after 4 h and reached stationary phase at 22 h. Notably, when compared with non-treated group, the biomass of S. mutans biofilm was increased with time from 0 to 72 h after treatment with 1.0, 2.5 and 5.0 mmol/L D-Trp. And at the same time point, the biomass was significantly less in each subgroup of treatment group than that of non-treated group (P<0.05). Crystal violet staining demonstrated that values of biomass(OD570)were less in treatment groups treated with 1.0, 2.5 and 5.0 mmol/L D-Trp than those of non-treated group (P<0.01). CLSM also showed that bacteria was adhered to the surface of media intreatment groups treated with 1.0, 2.5 and 5.0 mmol/L D-Trp. The values of biomass were lower in treatment groups than those of non-treated group (P<0.01). The MIC against S. mutans was 0.073 mg/L in both experimental group and negative control group. The values of MBIC were 0.293 mg/L and 2.344 mg/L in experimental group and negative control group, respectively. Under the action of 5.0 mmol/L D-Trp, the MBIC of S. mutans was reduced to 1/8. Conclusion Results indicate that D-Trp may inhibit the formation of S. mutans biofilm and promote the dispersal of biofilm already formed. D-Trp may further help CHX exert its bactericidal activity to S. mutans.