BACKGROUND: There are currently many cryopreservation methods for the aminotic membrane, which have varying effects on the ultrastructure and biological activity of amniotic membrane, but on no one is effective.
OBJECTIVE: To compare the effects of different cryopreservation methods on the ultrastructure and viability of aminotic membrane and to seek the ideal cryopreservation method.
METHODS: Aminotic membrane separated from the fresh placenta was preserved respectively with deep-frozen cryopreservation and vitrification, and everyway was run for 3 and 6 months. Fresh aminotic membrane was used as control. The ultrastructure of aminotic membrane was observed by transmission electron microscopy, and the viability of aminotic membrane was assessed by microcomputer analysis system for biological oxygen consumption, and immunohistochemical staining combined with image analysis system was used for lactate dehydrogenase activity.
RESULTS AND CONCLUSION:After 3 and 6 months of crypreservation, the damage to the ultrastructure of aminotic membrane by vitreous cryopreservation was slighter than that of amniotic membrane cryopreserved at-80℃. Compared with the fresh aminotic membrane, the gray value of lactate dehydrogenase and partial pressure of oxygen were significantly decreased in the cryopreserved aminotic membrane by deep-frozen cryopreservation at 3 and 6 months (P < 0.05) and by vitreous cryopreservation at 6 months (P < 0.05), but there was no statisticaly significant difference in the change rate of oxygen partial pressure and the gray value of lactate dehydrogenase between the fresh aminotic membrane and the cryopreserved aminotic membrane by vitreous cryopreservation at 3 months. The present study led to the conclusion that vitreous cryopreservation protocol alows to not only maintain the integrity of AM, but also to preserve the viability of the cels. So the vitreous cryopreservation is superior to the deep-frozen cryopreservation for cryopreservation of aminotic membrane.

Objective To obtain serological prevalence data for HBV markers in inpatients of Xi'an area with consequence of providing basis for nosocomial infection control and clinical stuff.Methods The serological markers of HBV (HBsAg,HBsAb,HBeAg,HBeAb,HBcAb) in serum of inpatients including 5 248 males and 5 345 females in 2015 were quantitatively detected by chemiluminescent analyzer ARCHITECT i4000SR.Results The infection rate of HBV was 7.01％ (743/10593) and there were 14 patterns of HBV serum markers in inpatients.Of all patterns of HBV infection in this study,there were 5.17 ％ (548/10 593) with HBsAg+ HBeAb+ HBcAb+,1.34 ％ (142/10 593) with HBsAg+ HBeAg+ HBcAb+,0.25％ (27/10 593) with HBsAg+HBcAb+ and 0.25％ (26/10 593) with other uncommon ones.Of all patterns of HBV convalescent stage,there were 21.02％ (2 227/10 593) with HBsAb+,13.71％ (1 452/10 593) with HBsAb+HBeAb+ HBcAb+,and 15.07％ (1 596/10 593) with HBsAb+HBcAb+.The percentage of five serum markers with negative was 31.38％ (3 324/10 593).There existed statistical difference for patterns of HBV serum markers concerning gender and different age groups,respectively (P＜0.05).The clinical departments with highest percentages of HBsAg-+-HBeAg+ HBcAb +-,HBsAg+ HBeAb+ HBcAb+ and HBsAb+ were department of gastroenterology with 7.39 ％ (36/487),department of gastroenterology with 16.43％ (80/487) and thoracic surgery one with 89.23％ (58/65),respectively.Conclusion This study provided clinical data of management and controlling the transmitting of HBV and promotion of HBV vaccination.Meanwhile it is necessary for government to take effective measures to reduce the infection rate of HBV in Xi'an area.

BACKGROUND: Models concerning tumor external environment mainly concentrated on laboratory two-dimensional culture and in vitro animal experiment, which lack of three-dimensional stereo.OBJECTIVE: To establish in vitro bone metastasis stereo models of human prostate carcinoma, and to investigate the effect of stem cells on proliferation rate and clustering size of prostate carcinoma cells. METHODS: Bone marrow mesenchymal stem cells (BMMSCs) were extracted from 2 clean grade SD rats. Alginate was used to simulate medullary microenvironment, where prostate carcinoma cells and BMMSCs were co-culturedd. Growth of the cells in the three-dimensional model was observed through microscope and histological sections. The carcinoma cells were transfected with green fluorescent protein. The proliferation of monoclonal cells clustering was observed under light microscope and fluorescence microscope. RESULTS AND CONCLUSION: In the co-culture group, the clustering speed, clustering amount and tumor formation rate were greater that those of the control group. The monoclonal cells clustering was formed at 7.75 days and 6.00 days in the control and co-culture groups, respectively, with cell counts of (95.13±11.63) and (112.53±14.67) after 10 days. The formation rate of fluorescent cell clones was (77.10±6.85)% in the control group and (64.55±6.21)% in the co-culture group, the difference had significance. The results suggested that: the alginate microenvironment is conductive to proliferation and clustering of prostate carcinoma cells and BMMSCs.

Objective To investigate the effects of caudal type homeobox 2 (Cdx2) silence on reservion of multi-drug resistance of gastric carcinoma cisplantin-resistant cell SGC7901/DDP.Methods Gastric carcinoma cisplantin-resistant cells SCG7901/DDP in the logarithmic phase were cultured in the plate,and were divided into the experimental group [gastric carcinoma cells of SGC7901/DDP were infected with a silent Cdx2-recombinanted lentiviral vector (pLL-Cdx2-shRNA)],the negative control group (gastric carcinoma cells of SGC7901/DDP were infected with empty lentiviral vector) and the blank control group (gastric carcinoma cells of SGC7901/DDP were not treated).The protein and mRNA expressions of Cdx2 and apoptosis related genes like c-myc,cyclin D1 and survivin were detected by the Western blot and reverse-transcription PCR,respectively.The sensitivity of the cells in the 3 groups to adriamycin,5-fluorouracil and cisplatium were assessed by MTT.The pump-out rate of adriamycin,cell cycle distribution and apoptosis of the 3 groups were analyzed using flow cytometry.All measurement data were expressed with mean ± standard deviation.Comparison among multi-groups was done by one-way analysis of variance,and comparison between 2 groups was done by SNK-q test.The enumeration data were analyzed using the chi-square test.Results The relative protein expression levels of Cdx2,c-myc,cyclin D1 and survivin were 0.187 ± 0.060,0.086 ± 0.004,0.016 ± 0.005 and 0.276 ± 0.012 in the experimental group,0.535 ± 0.033,0.379 ± 0.006,0.141 ± 0.003 and 0.672 ± 0.009 in the negative control group,and 0.567 ± 0.014,0.354 ± 0.004,0.162 ± 0.008 and 0.517 ± 0.313 in the blank control group,respectively.The relative protein expression levels of Cdx2,c-myc,cyclin D1 and survivin in the experimental group were significantly lower than those in the negative control group and the blank control group (F =247.385,3.353,597.882,98.628,P ＜0.05).The relative mRNA expression levels of Cdx2,c-myc,cyclin D1 and survivin were 0.184 ± 0.010,0.212 ± 0.022,0.045 ± 0.009 and 0.401 ± 0.027 in the experimental group,0.894 ± 0.056,0.538 ± 0.021,0.163 ±0.009 and 0.824 ± 0.016 in the negative control group,and 0.837 ±0.049,0.545 ±0.032,0.157 ±0.010 and 0.782 ±0.056 in the blank control group,respectively.The relative mRNA expression levels of Cdx2,c-myc,cyclin D1 and survivin in the experimental group were significantly lower than those in the negative control group and the blank control group (F =243.776,161.793,138.523,118.426,P ＜ 0.05).The IC50 values detected by MTT of adriamycin,5-flurouracile and cisplatin to gastroc carcinoma cisplantin-resistant cell SCG7901/DDP were (0.12 ± 0.05) mg/L,(0.52 ± 0.13) mg/L and (0.82 ± 0.13) mg/L in the experimental group,(0.33 ± 0.08) mg/L,(4.10.± 1.25) mg/L and (2.81 ± 0.50) mg/L in the negative control group,(0.39 ±0.15)mg/L,(4.05 ± 1.44) mg/L and (3.28 ± 1.03) rng/L in the blank control group,respectively.The pump-out rates of adriamycin of the experimental group,negative control group,and the blank control group were0.21％,0.37％ and 0.35％.Compared with the negative control group and the blank control group,the IC50values of adriamycin,5-fluorouracil and cisplatin in the experimental group were significantly increased,and thepump-out rate of adriamycin was significantly decreased (F =8.101,13.854,15.159,x2 =7.106,P ＜ 0.05).The ratios of cells in the G0/G1 phase were 17.87％,34.71％ and 37.20％ in the experimental group,negative control group and the blank control group,respectively.Compared with the negative control group and blank control group,the ratio of cells in the G0/Gt was significantly decreased (x2=1.055,P ＜ 0.05).The ratio of cells in the G2/M phase in the experimental group was 11.93％,and the apoptosis rate was 31.13％,which were significantly higher than the negative group (0.26％,16.58％) and the blank control group (0.35％,13.18％) (x2=2.249,11.030,P ＜ 0.05).Conclusions Silent Cdx2 can effectively enhance the sensitivity of the SGC7901/DDP cells and the intracellular accumulation concentration of the drugs.Silent Cdx2 can also reverse the multidrug resistance of the SGC7901/DDP cells.

OBJECTIVETo compare the therapeutic effect and toxicity of chemotherapy, used alone or in combined with Shenqi Fuzheng Injection (SFI), for the treatment of advanced colorectal carcinoma (ACRC).

METHODSOne hundred and fifty-two patients with ACRC were equally randomized by digital table, to the treated group, treated by chemotherapy of FOLFOX regimen combined with SFI, and the control group treated by FOLFOX regimen alone. The therapeutic effect and adverse reaction of the treatment in patients were assessed.

RESULTSThe effective rate (CR +PR) was 63.2% (48/76) in the treated group and 46.1% (35/76) in the control group, showing significant difference between the two groups (P < 0.05). The median survival time in the two groups was 31 weeks and 28 weeks respectively. CD4/CD8 ratio was significantly increased in the treated group (1.56 +/- 0.21, 1.64 +/- 0.28, P < 0.05), but significantly decreased in the control group (1.58 +/- 0.22, 1.46 +/- 0.33, P < 0.01). Quality of life in the former group was higher than that in the latter group (P < 0.05). Times/case of nausea, vomiting, leukopenia occurring in the control group was more than those in the treated group A (P < 0.05).

CONCLUSIONBy combining with SFI, some adverse reactions of chemotherapy (such as nausea, vomiting, leukopenia) and its influence on patients' immunity could be alleviated in treating ACRC, which might enhance the efficacy of chemotherapy, and improve the quality of life and prolong the median survival time in patients.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Colorectal Neoplasms ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Fluorouracil ; therapeutic use ; Humans ; Leucovorin ; therapeutic use ; Organoplatinum Compounds ; therapeutic use ; Quality of Life ; Survival Rate

OBJECTIVETo investigate the effects of E2F-1 gene silencing on the chemosensitivity of human gastric cancer SGC-7901/DDP cells to cisplatin and explore the underlying mechanism.

METHODSGastric cancer SGC-7901/DDP cells were transfected with the recombinant lentivirirus vector Lv-shRNA-E2F-1 for E2F-1 gene silencing, with cells transfected with the control recombinant lentivirirus vector Lv-shRNA-NC as the negative control. MTT assay was used to evaluate cisplatin chemosensitivity of the cells, and the cell apoptosis rate and cell cycle distribution were detected by flow cytometry. The mRNA and protein expressions of E2F-1 and apoptosis-related genes (survivin and Bcl-2) were detected by RT-PCR and Western blotting.

RESULTSMTT assay showed that the IC50 of cisplatin was significantly lowered in Lv-shRNA-E2F-1-transfected cells compared with the negative and blank control cells (P<0.05). Lv-shRNA-E2F-1 transfection caused significant cell cycle arrest in G0/G1 phase and induced obvious cell apoptosis. Compared with Lv-shRNA-NC group and the blank control group, Lv-shRNA-E2F-1 group showed significantly lowered expressions of E2F-1 mRNA by 45.0% and 41.3% and E2F-1 protein by 66.7% and 70.5%, survivin mRNA by 30.3% and 28.7% and survivin protein by 56.5% and 53.6%, and Bcl-2 mRNA by 76.6% and 76.8% and Bcl-2 protein by 74.6% and 79.9%, respectively. No significant difference was found in the measurements between Lv-shRNA-NC group and the blank control group (P>0.05).

CONCLUSIONE2F-1 gene silencing can enhance cisplatin chemosensitivity of gastric cancer SGC-7901/DDP cells possibly by down-regulating survivin and Bcl-2 expressions, suggesting the value of E2F-1 as a new chemotherapeutic target for gastric cancer.

Apoptosis ; Cell Cycle ; Cell Line, Tumor ; drug effects ; Cisplatin ; pharmacology ; Down-Regulation ; E2F1 Transcription Factor ; genetics ; metabolism ; Gene Silencing ; Genetic Vectors ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; RNA, Small Interfering ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

To study the effect of isoprenoid and aliphatic saturated alcohols as modificator on benzoic nitrogen mustard, the intermediate 4-[N,N-bis(2-chloroethyl) amino] benzoic acid 4 was prepared in four steps utilizing p-amino benzoic acid as the starting material. Target compounds were synthesized by the catalytic esterification of DCC/DMAP and the structures of the six new esters were characterized by elemental analysis, 1H NMR, 13C NMR and MS. Antitumor activities were evaluated in vitro using MTT assay. The result showed that some derivatives were more potent than the intermediate 4, and compound 5c modified with dodecanol exhibited similar activity to the commercial drug melphalan.
Aminobenzoates ; chemical synthesis ; pharmacology ; Animals ; Antineoplastic Agents, Alkylating ; chemical synthesis ; pharmacology ; CHO Cells ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cricetinae ; Cricetulus ; Humans ; Inhibitory Concentration 50 ; K562 Cells ; Melanoma, Experimental ; pathology ; Melphalan ; pharmacology ; Nitrogen Mustard Compounds ; chemical synthesis ; pharmacology

OBJECTIVETo explore the association of plasma level of advanced oxidation protein products (AOPPs) with Framingham risk score in type 2 diabetic patients without vascular diseases.

METHDOSThis cross-sectional study was conducted among type 2 diabetic patients without vascular diseases recruited from 3 affiliated hospitals of Southern Medical University between March, 2010 and May, 2011, with age- and gender-matched healthy individuals as the control group. The demographic data were collected from all the participants, and the biochemical indexes and plasma levels of AOPPs were examined. The risk of cardiovascular disease in 10 years was assessed for all the participants based on their Framingham risk scores.

RESULTSA total of 112 diabetic patients and 49 healthy subjects were enrolled in this study. The diabetic patients had significantly higher body mass index (BMI), blood glucose, triglyceride, low-density lipoprotein (LDL), plasma AOPPs and Framingham risk score but lower high-density lipoprotein level than the control subjects. Spearman correlation analysis showed that plasma level of AOPPs was positively correlated with the Framingham risk score (r=-0.44, P<0.001), and further multiple linear regression analysis suggested that plasma AOPPs level was independently correlated with the Framingham risk score (β0.305, P<0.001).

CONCLUSIONType 2 diabetic patients without vascular diseases have significantly higher plasma levels of AOPPs and are at a greater risk of cardiovascular events in 10 years than healthy individuals. Plasma AOPPs are positively correlated with the Framingham risk score, suggesting the value of plasma AOPPs level in predicting the risk of cardiovascular events in these patients.

Objective To investigate the relationship between serum uric acid level and renal function decline by retrospective cohort study.Methods Through the physical examination system of the First People's Hospital of Foshan,the physical examination data from 2015 to 2018 of a public institution in Foshan city were obtained.The gender,age,blood cell analysis,liver function,serum creatinine,uric acid,fasting blood glucose were obtained.The change of eGFR (△eGFR=eGFR2018-eGFR2015) was analyzed.Results A total of 2505 subjects were followed up for four years.The subjects were divided into △eGFR ≥0 group and △eGFR ＜ 0 group.There were 845 subjects in △eGFR ≥0 group,and 1660 subjects in △eGFR ＜ 0 group.Compared with that in △eGFR ＜ 0 group,the base-level of uric acid in △eGFR ≥ 0 group was higher [(349.48±87.62) μmol/L vs (325.72±82.58) μmol/L,t=6.669,P ＜ 0.001],but the rate of uric acid decline was greater [-15.00(-53.50,17.00) μmol/L vs 15.50(-18.00,49.00) μmol/L,Z=-13.470,P ＜ 0.001].According to the levels of uric acid in 2015 and 2018,then the subjects were divided into four groups,normal to normal group (N-N,1551 cases),normal change into high uric acid group (N-H,299 cases),high uric acid drop to normal group (H-N,238 cases),and high to high uric acid group (H-H,417 cases).The △eGFR was-1.58(-4.17,1.01) ml · min-1 · (1.73 m2) 1 in N-N group,and-3.60(-7.24,-0.98) ml · min-1 · (1.73 m2)-1 in N-H group,-0.20(-3.14,3.27) ml· min-1· (1.73 m2)-1 in H-N group,-0.96(-4.07,1.93) ml· min-1· (1.73 m2)-1 in H-H group,respectively.The △eGFR decreased most significantly in N-H group than the other three groups (x2=103.130,P ＜ 0.001).Multivariate logistic regression analysis showed that elevated uric acid was an independent risk factor for eGFR decline (OR=1.739,95％CI 1.587-1.906,P ＜ 0.001),while elevated indirect bilirubin (OR=0.968,95％CI 0.943-0.993,P=0.013),elevated red blood cells (OR=0.815,95％ CI 0.680-0.976,P=0.026) were independent protective factors for eGFR decline.Conclusion Elevated uric acid is an independent risk factor for the decline of renal function.Good control of hyperuricemia is beneficial to the protection of renal function.

Objective Sperm screening is an essential step in IVF procedures. The swim-up method, an assay on sperm motility, is used clinically to select the ideal sperm for subsequent manipulation. However, additional parameters, including acrosome reaction capability, chemotaxis, and thermotaxis are also important indicators of mammalian sperm health. To monitor both sperm motility and chemotaxis simultaneously during sperm screening, we designed and constructed a microdevice comprising a straight channel connected with a bi-branch channel that mimics the mammalian female reproductive tract. Methods The width and length of the straight channel were optimized to select the motile sperm. Cumulus cells were selectively cultured in the bi-branch channel to generate a chemoattractant-forming chemical gradient. Sperm chemotaxis was represented by the ratio of the sperm swimming towards different branches. Results The percentage of motile sperm was improved from ( 58. 5 ± 3. 8 ) % to ( 82. 6±2.9)% by a straight channel 7 mm in length and 1 mm in width. About 10% of sperm were found chemotactically responsive in our experiment, which is consistent with previous studies. Conclusion The combined evaluation of both sperm motility and chemotaxis was achieved for the first time, and the motile and chemotactically responsive sperm can be easily enriched on a lab-on-a-chip device to improve IVF outcome.