Objective To optimize the manufacture process for Zhibai Anshen oral liquid. Methods The orthogonal designed experiments were conducted to monitor the effects of three factors on the content of mangiferin. The three factors included the amount of water, extraction time and alcohol precipitation concentration. Six month accelerated stability study and twelve month long term stability study were performed. Results Optimum percolation process was boiling the mixture with 10 times of water for 1 hour, followed by deposition with 60% alcohol. Conclusion This optimized process can be used for mass production.

Objective To analyze the changes of endoplasmic reticulum stress (ERS) in the spleen of water-improving fluorosis rat,to explore the mechanism of fluoride-induced immune system damage,and to provide a scientific basis for prevention and control of endemic fluorosis.Methods Forty-eight male Wistar rats of SPF grade were randomly divided into control group and low,medium and high fluoride dose groups according to body mass (120-140 g),12 rats in each group.The sodium fluoride (NaF) content was 0,50,100 and 150 mg/L,respectively.The animals were allowed free access to water and food.After 12 weeks of fluoride exposure,6 rats in each group were selected to isolate the spleen;the remaining rats in each group were changed to drink distilled water containing no NaF,and the spleen was separated after 12 weeks of feeding.The levels of mRNA of glucoseregulated protein (GRP78),spliced X-box binding protein 1 (XBP1-s),activating transcription factor 4 (ATF4),homologous protein (CHOP) and cysteine containing aspartate specific protease 12 (Caspase-12) in spleen were determined by quantitative real-time PCR (qRT-PCR).Results Before the water-improving,the expressions of GRP78 (1.00 ± 0.09,1.69 ± 0.35,1.39 ± 0.29,1.19 ± 0.19),XBP1-s (1.00 ± 0.12,1.40 ± 0.23,1.24 ± 0.26,1.38 ± 0.11),ATF4 (1.00 ± 0.17,1.86 ± 0.56,2.33 ± 0.55,1.95 ± 0.74),CHOP (1.00 ± 0.53,2.84 ± 0.68,3.06 ± 1.29,2.50 ± 0.35) and Caspase-12(1.00 ± 0.12,1.90 ± 0.29,1.56 ± 0.35,1.76 ± 0.23) mRNA in the control group and low,medium and high fluoride dose groups were statistically significant (F =8.45,5.38,6.38,8.21,11.31,P ＜ 0.05).Except for the GRP78 in high fluoride dose group,the above indicators in fluoride groups were higher than the control group (P ＜ 0.05).After the water-improving,the expressions of GRP78 (1.00 ± 0.36,0.75 ± 0.13,0.98 ± 0.41,0.47 ± 0.19),XBP1-s (1.00 ± 0.25,0.70 ± 0.06,0.74 ± 0.17,0.65 ± 0.21),ATF4 (1.00 ± 0.51,0.66 ± 0.09,0.91 ± 0.34,0.81 ± 0.29),CHOP (1.00 ± 0.36,0.92 ± 0.12,0.84 ± 0.16,0.67 ± 0.20) and Caspase-12 (1.00 ± 0.45,0.65 ± 0.11,0.65 ± 0.25,0.51 ± 0.27) mRNA in the control group and low,medium and high fluoride dose groups were not statistically significant (P ＞ 0.05).Before and after the water-improving,the expressions of XBP1-s,ATF4,CHOP and Caspase-12 mRNA were statistically significant in fluoride groups (P ＜ 0.05),and the GRP78 only had a statistically significant difference in the low fluoride dose group (P ＜ 0.05).Conclusions Fluoride exposure causes ERS response in rat spleen,up-regulation of ERS-related gene expression,which is decreased after water-improving,and the ERS response is weakened.The water-improving may contribute to the recovery of fluoride-induced immune function damage.

Objective To study the effects of different fluoride concentrations on proliferation and apoptosis of rats' immune cells,and to explore the effects of fluoride on immune function.Methods Sixty male Wistar rats were randomly divided into four groups,and the concentrations of NaF in drinking water for each group (15 rats in each group) were 0,50,100,and 150 mg/L,respectively.They were freely fed water and commercial standard chow.All rats were sacrificed after 12 weeks.Thymus index was calculated.Lymphocyte proliferation activity in the blood was detected by Cell Counting Kit-8 assay.The apoptosis of monocytes in blood was detected by Annexin V/PI method.The apoptosis of lymphocytes in spleen and thymus was detected by Tunal method.Results After 12 weeks of fluoride exposure,the difference in the thymus immune organ index between the four dose groups was statistically significant (F =6.50,P ＜ 0.05);after 12 weeks,the thymus immune organ index of the low and middle dose groups was significantly lower than that of the control group (0.70 ± 0.19,0.84 ± 0.18 vs 1.16 ± 0.33,P ＜ 0.05).There were significant differences in B and T lymphocyte viability between the four dose groups (F =539.97,4.92,P ＜ 0.05).The viability of B lymphocyte in the blood of the middle dose group was significantly lower than those of control group and the low dose group [(58.09 ± 4.59)％ vs (100.00 ± 9.01)％,(106.70 ± 4.82)％,P ＜ 0.05].The viabilities of T lymphocyte in the blood of the low and middle dose groups were significantly lower than that of the control group [(81.11 ± 2.93)％,(75.68 ± 2.34)％ vs (100.00 ± 34.02)％,P ＜ 0.05].The apoptosis rates of blood mononuclear cells in the fluoride-treated groups were significantly increased than that of control group [(48.00 ± 7.45)％,(47.26± 5.94)％,(48.20 ± 3.40)％ vs (32.50 ± 13.70)％,P ＜ 0.05).Immunohistochemistry results showed that the number of apoptotic lymphocytes in the spleen and thymus increased significantly in high dose group.Conclusion Fluoride can reduce the thymus immune organ index of rats,affect the proliferation of lymphocytes in blood,thymus and spleen,and promote apoptosis of lymphocytes,thus affecting the immune function.

Objective To investigate the pathogenic mutations of BRCA1 and BRCA2 in patients with early-onset breast cancer(≤35 years) and explore the relationships between BRCA1/2 mutations and clinical features.Methods Seventy-four patients with early-onset breast cancer were enrolled,who were treated in Hospital 307 between September 2014 and June 2016.High-throughput sequencing was used to test the 49 exon sequences and adjacent sequences of BRCA1 and BRCA2.χ2 test was used to analyze the distribution of BRCA1/2 pathogenic mutations in each group that was set up according to clinical features.Results Fifteen mutations(20.27%) were identified,including 5(6.76%) in BRCA1 and 10(13.51%) in BRCA2.Eleven new pathogenic mutations were discovered,and BRCA1:c.5470_5477delTGCCCAAT was found in one patient.The frequency of BRCA1/2 mutations in the group with a family history of breast cancer or ovarian cancer was higher than in the group without a family history (40.91% vs 11.54%) (χ2=6.534,P=0.011).Conclusion BRCA1/2 pathogenic mutation is significant for early-onset breast cancer,especially for those with a family history of breast or ovarian cancer.The new mutations may be specific to Chinese people.BRCA1:c.5470_5477delTGCCCAAT may be the ancestor mutation among the Chinese.

Objective To establish a quality control standard for Qingre Baidu granules .Methods Isatidis Radix ,Fruc-tus Forsythiae ,Herba Violae ,and Glycyrrhizae were identified by TLC ,and the concentration of chlorogenic acid was deter-minedbyHPLC.ThismethodwasemployedonanAgilentZORBAXSB-C18column(4.6mm×250mm,5μm)at30℃ witha mobile phase of acetonitrile (A) and 0 .2% formic acid (B) using the gradient elution program shown as follows :0-12 min , 11%-12% A run at the flow rate of 1 .0 ml/min .The injection volume was 20 μl and the detection wavelength was 327 nm . Results Characteristic spots could be detected by TLC and the specificity of the method was satisfactory .As for chlorogenic acid ,the equation of linear regression of chlorogenic acid was Y=60 .239 4X+9 .096 3 (r=0 .999 9) with the linear range of 6.19-396 .00 μg/ml .The average recovery was 99 .66% (RSD=2 .82% ) .Conclusion The established method is simple ,reli-able ,reproducible ,and can be used for the quantitative determination and quality control of Qingre Baidu granules .

This paper is to evaluate the biocompatibility and cytotoxicity of a new Ni-free Zr-based bulk metallic glass (BMG), Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3, by comparing it with conventional Ti6Al4V alloy. According to ISO 10993-5: 1999 and GB/T 16886.5-1997 standards, Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3, pure Zr and Ti6Al4V materials were extracted with surface area of sample/volume of medium ratio being 1 cm2/mL and 0.5 cm2/mL, respectively. The viabilities of MG-63 cells (Human osteosarcoma cell line) cultured in the BMG medium extracts for 1, 3 and 5 days were determined by CCK-8 assay. The cellular morphology of MG-63 cells cultured on the surface of samples for 3 days was tested through laser scanning confocal microscopy (LSCM) and scanning electron microscopy (SEM). The relative growth rate (RGR) of MG-63 cells cultured in Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3 and pure Zr were both more than 85%, indicating that the cytotoxicity of BMG was relatively low and met the national biomedical material eligibility standard. There was insignificant difference in the morphology of MG-63 cells cultured in the BMG medium extracts and the control group through LSCM and SEM, which showed the BMG had excellent biological compatibility. The Zr-based bulk metallic glass Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3 and the conventional Ti6Al4V alloy both had no obvious cytotoxicity to MG-63 cells. These results provided evidence that the new Zr-based bulk metallic glass could be potential replacement material for the orthopedic surgical implant.
Alloys ; Biocompatible Materials ; Cell Line ; Glass ; Humans ; Microscopy, Electron, Scanning ; Nickel ; Titanium ; Zirconium

This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis

Objective To analyze the correlation between the expression of Ki-67 and Her-2 and clinical features in breast invasive ductal carcinoma .Methods Clinical and pathological data of 202 female breast invasive ductal carcinoma patients were collected between January 2012 and September 2014 .The correlation between the expression of Ki-67 and Her-2 and clinical features was analyzed .Results The positive rate of Ki-67 was 73.3%(148/202) and that of Her-2 was 19.3(39/202).The expression of Ki-67 was related to histological grading , ER and PR status(P<0.05),but not to age, tumor size or lymph node status(P>0.05).The expression of Her-2 was related to ER and PR status (P<0.05), but not to age, tumor size, histological grading, or lymph node status(P>0.05).Moreover, the low expression Ki-67 and negative expression of Her-2 were positvely corelated with the positive expression of hormone receptor [estrogen receptor(ER),pro-gestin receptor(PR)](P<0.05).Conclusion High expression of Ki-67 suggested that the differential grade of tumor was lower , malignant grade was higher , infiltration was stronger and metastasis was easier .High expression of Ki-67 and positive Her-2 suggests lower sensitivity to endocrine therapy and treatment .Detection of Ki-67 and Her-2 expression in breast inva-sive ductal carcinoma is of guiding significance for understanding the degree of malignancy and for evaluating prognosis .

Objective To investigate the effects of 8-week Grade 2 and 5 Aerobic Dances on physical fitness and visual evoked potentials(VEPs)of female students.Methods ① Forty five female student volunteers were randomly divided into three groups:control group(C).Grade 2(G2)and 5(G5)Aerobic Dances groups.The aerobic dances groups took part in Chinese National Aerobic Dance Grade 2(135 tempo/min)and 5(148 tempo/min)separately,lh/day,3 days/week,for 8 weeks.②Physical fitness in three groups was compared before and after 8 weeks.③The NDI-200 neural electricity tester(Haishen)was applied to measure the PR-VEPs in all subjects before and after the experiment,and the changes in VEPs latencies,inter-peak latencies and peak to peak amplitudes were compared.ResuIts①The number of sit-ups with 90°knees flexion and sit-and reach,and maximuin oxygen uptake of both groups G2 and G5 increased significantly(P<0.01),especially in group G5.②As compared with the pre-test,the VEPs latencies in both groups G2 and G5 shogened,and the VEP P100 latency shortened more significantly in group G5(P<0.01).③No significant differences were found in both the VEPs inter-peak latencies and peak to peak amplitudes among the three groups(P>0.05).Conclusion Both Grade 2 and 5 Aerobic Dances could improve the physical fitness and shorten the VEP P_(100) latency,especially the Grade 5 Aerobic Dance.

AIM To study the pharmacokinetic characters of dauricine(Dau) in rats after different administration ways. METHOD RP-HPLC method was used in the study. RESULT The results indicated that the plasma C-T curve conform to two-compartment open model after iv. The plasma concentration of Dau in rats after ig Dau 150 mg*kg-1 is low, less than 1 mg*L-1 of peak concentration. The absolute bioavailibility is about 16.6 %. The plasma concentration-time profile shows a double-peak phenomenon. The time taken to reach the peak is about 15 min after ig and the trough time is 3 h. The plasma concentration increased again in 4 h to form the second peak. The studies suppose a stomach-intestine recirculation of Dau is the major reason for double-peak phenomenon. Dau has a wide distribution in rat body. It lies in all tissues and organs in both adminastration ways. The tissue Dau concentration are hundreds times higher than that in plasma concurrently. Feces is the main route whereby Dau are excreted from the rats after ig 150 mg*kg-1. The excreted percentage through feces is 26.29 %, while through urine is 4.93%. The total amount is 31.22% after 48h of oral administration of Dau. The study of the mean percentage of the dose remaining in stomach, small intestine, large intestine and whole GI tract from each rat sacrificed at different times after oral administration of Dau suggest the stomach-intestine circle. CONCLUSION　The bioavailibility of Dau is low. The plasma drug concentration versus time curve shows an innormal double-peak phenomenon. Dau can distribute abroadly to almost all kinds of the tissues in rats. The main excretion routes are through feces and urine. The pilot study suggests that stomach-intestine circle be the main reason for the innormal double-peak phenomenon.