Objective To determine the content of baicalin in Radix Scutellariae granule from different manufacturers using near infrared spectroscopy(NIRS)technology. Methods Utilizing NIRS combined with partial least squares, and simultaneously optimizing the pretreatment methods and the range of spectrum, the quantitative model of baicalin in Radix Scutellarie granules was established. Results In the model correlation coefficient(R2)was 0.9702, root-mean-square error of calibration(RMSEC)was 0.555, and root-mean-square error of predict(RMSEP)was 1.05. Conclusion In this paper, rapid quality analysis method of Radix Scutellariae granules from different manufacturers is studied using NIRS technology. The model can predict the content of baicalin nondestructively and rapidly. It can give some reference for researching the quality control of Radix Scutellariae granules.

BackgroundCorneal graft reject is a major cause of corneal transplantation failure.Although many immune-suppressing drugs have been utilized to reduce the reject response,their adverse effects on organ and tissue are still insoluble.The tolerance induction of natural killer T (NKT) cells is currently under investigation.However,the study on the application of NKT cells in high risk corneal transplantation is seldom.ObjectiveThe present study was to explore the effects of α-GalCer-activated NKT cella on allografts survival after high-risk corneal transplantation surgery via retro-bubar injection.Methods The lymphocytes were picked up from the spleen of SPF Lewis rats and cultured in RPMI 1640 medium with 100 mg/L α-GalCer.After one week,NKT cells were sorted by the FACSVantage system as CD161+ TCR-α+ cell from the lymphocytes with the cell densities 5×106/ml.Ten SPF Fisher344 rats were used to prepare the donor corneas,and 20 Lewis rats served as recipients.The high risk corneal transplantation models were created by corneal suturing in 20 recipient rats.Penetrating keratoplasty (PKP) was performed in the model rats.0.1 ml NKT cells or the same volume of normal saline solution were retro-bubarly injected at the end of surgery respectively.The corneal allografts were observed and scored based on Holland criteria at the three-day interval under the slit lamp for 30 days.Two weeks after surgery,three rats from each group were sacrificed by excessive anesthesia method and the eyeballs were obtained for histopathological examination.The inflammatory cell infiltration ( CD4+ and CD8+ ) in grafts was evaluated by immunochemistry and flow cytometry.The use of the animals complied with the Statement of ARVO.ResultsThe mean survival time of the allografts was (7.90± 1.37) days in normal saline solution group and (14.70± 1.49) days in NKT cell group,showing a statistically significant difference between the two groups ( t =10.61,P =0.00 ).Two weeks after surgery,all the allografts showed the severe opacity with lots of new blood vessels and edema in normal saline solution group.However,the corneal grafts were clear in NKT cell group.Abundant CD4+ and CD8+T lymphocytes were seen in the allografts in normal saline solution group,but the inflammatory cells were obviously less in NKT cell group.The percentage of NKT cells in the spleen was (5.67±0.25)％ in NKT cell group and ( 1.21±0.19)％ in normal saline solution group ( t =8.43,P =0.00 ).Conclusionsα-GalCer-activated NKT cells can prolong the survival time of allografts in high-risk corneal transplantation.Retro-bubar injection of α-GalCer-activated NKT cells probably is a new approach to the prevention of the rejection of corneal transplantation.

OBJECTIVEThe study aimed to test if Paridis Rhizoma total saponins (PRTS) could induce apoptosis of human gastric cancer cell MKN-45.

METHODBased on the previous researches, PRTS was set by different concentrations to treat human gastric cancer cell for 12 h (5, 10, 20 mg x L(-1)). Fluorescent staining methods were adopted to observe apoptotic morphological changes of MKN-45. The apoptosis rates were analyzed by flow cytometry with Annexin V-FITC/PI staining. The enzymatic activities of caspase-3 and caspase-8 were measured by ELISA. The protein levels of Fas and FasL were detected by Western blotting.

RESULTUnder a fluorescence microscope, MKN-45 treated by PRTS was seen typical apoptotic morphological features. PRTS significantly increased the rate of apoptosis. Compared with the control group, there exsited significant differences in apoptosis rate of PRTS concentration of 20 mg x L(-1) (P < 0.01); besides, the enzymatic activities of caspase-3 and caspase-8 were promoted obviously after the effect of PRTS on MKN-45 cells for 12 h (P < 0.01). The protein levels of Fas and FasL in the MKN-45 were upgraded significantly.

CONCLUSIONPRTS can induce apoptosis of human gastric cancer cell MKN-45 , which is concerned with caspase-3 and caspase-8 and upgraded Fas and FasL.

Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 8 ; genetics ; metabolism ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; metabolism ; Humans ; Magnoliopsida ; chemistry ; Rhizome ; chemistry ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Stomach Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; fas Receptor ; metabolism

Objective To evaluate the clinical value of contrast-enhanced ultrasound (CEUS) in preoperative diagnosis of anal fistula. Methods Forty-five patients with fistula in ano were evaluated by physical examination, then CEUS were peformed by injecting SonoVue through the external opening to enhance the detection of the fistulous track and the internal opening. The results of CEUS were matched with surgical features to establish their accuracy in preoperative assessment of anal fistula. Results Simple typing fistula was found in 19 of 45 patients and 26 patients had complex fistulas. The accurate diagnostic rates by conventional ultrasound of simple typing fistula and complex fistulas were 89. 5% and 61.5%,respectively,the accurate diagnostic rates by CEUS of simple typing fistula and complex fistulas were 94. 7% and 92.3%, respectively, there was no significant difference between conventional ultrasound and CEUS about simple typing fistula( P ＞0.05), and there was significant difference about complex fistulas ( P ＜0. 05). Conclusions CEUS has a good visibility and accurate rate for diagnosis of anal fistula,It plays an important role for operation.

Akt1 is a serine-threonine protein kinase that has been implicated in the control of cellular metabolism,survival and growth.Elevated expression of Akt1 has been noted in a significant percentage of human tumors,promoting cellular metastasis.Conversely,some studies have revealed hyperactivated Akt1 inhibited the invasiveness and metastasis of breast cancer cells.To clarify the definite effect of Akt1 on tumorigenesis and development,Akt1 was silenced by RNAi in the highly metastatic murine breast cancer 4T1 cells.Akt1 silencing didn't affect the proliferation of breast cancer cells in MTT assay,while reduced the migration in Transwell assay.Consistent with the above results,Akt1 silencing didn't change the primary tumor weight,but significantly suppressed lung metastasis of 4T1 cells.These observations indicated Akt1 plays an important role in murine breast cancer metastasis,and suggested that Akt1 might be a therapeutic target for breast cancer metastasis.

BACKGROUNDElevation of intraocular pressure is usually associated with primary open angle glaucoma and caused by increased outflow resistance. A two-color fluorescent tracer technique was developed to investigate the hydrodynamics of aqueous humor outflow with changing intraocular pressure within the same eye, to better understand the relationship between outflow facility and effective filtration area.

METHODSEighteen enucleated bovine eyes were first perfused at 30 mmHg with Dulbecco's phosphate-buffered saline containing 5.5 mmol/L D-glucose. After a stable baseline facility, red fluorescent microspheres (0.5 microm, 0.002% v/v) were exchanged and perfused. Eyes in the one-color control group (n = 6) were immediately perfused with fixative. In the experimental group (n = 6), eyes were perfused with green tracer after intraocular pressure reduced to 7 mmHg, while in the two-color control group (n = 6), eyes were perfused with green tracer with intraocular pressure remaining at 30 mmHg. All 12 eyes were then perfusion-fixed. Outflow facility was continuously recorded in all eyes. Confocal images were taken along the inner wall of the aqueous plexus and the percent of the effective filtration length (PEFL; length of inner wall exhibiting tracer labeling/total length of inner wall) was measured. The relationships between outflow facility and PEFL were analyzed statistically.

RESULTSNo significant differences were found in baseline facilities (microl x min(-1) x mmHg(-1)) among the three groups (the experimental group: 0.93 +/- 0.12; the two-color control group: 0.90 +/- 0.19; the one-color control group: 0.98 +/- 0.13). In the experimental group, the outflow facility was significantly higher at 7 mmHg (4.29 +/- 1.01) than that at 30 mmHg (1.90 +/- 0.67, P < 0.001), which corresponded to a significant increase in the PEFL at 7 mmHg (54.70 +/- 8.42) from that at 30 mmHg ((11.76 +/- 4.56)%, P < 0.001). The PEFL labeled by red fluorescent microspheres in the experimental group ((11.76 +/- 4.56)%) showed no significant difference from that of the one-color control group ((13.39 +/- 2.19)%, P = 0.473) or the two-color control group ((11.49 +/- 4.95)%, P = 0.930). The PEFL labeled by green fluorescent microspheres in the experimental group ((54.70 +/- 8.42)%) was significantly higher than that of the two color control group ((37.34 +/- 8.17)%, P = 0.010). A positive correlation was found between outflow facility and PEFL (r = 0.897, R(2) = 0.804) in the experimental group.

CONCLUSIONSChanges in aqueous humor outflow patterns before and after a change in intraocular pressure can be successfully distinguished within the same eye using our newly developed two-color tracer perfusion technique. The PEFL showed positive correlation with the outflow facility.

Animals ; Aqueous Humor ; physiology ; Cattle ; Intraocular Pressure ; Luminescent Proteins ; metabolism ; Microscopy, Confocal ; Microspheres ; Perfusion ; methods

Objective To investigate the sanitation status of centralized air ventilation system in Putuo District of Shanghai, to analyze correlation between different indexes and to provide basis for improving the health status of public places and strengthening health supervision. Methods According to the requirements of WS 394-2012 The Sanitation Criterion of the Centralized Air Ventilation System in Public Places, 18 centralized air ventilation systems in public places in Putuo District of Shanghai were randomly selected for sampling, testing and evaluation during 2014-2018, and 9 indexes of air supply, inner surface of air duct and indoor air were analyzed for correlation. Results The overall qualified rate of centralized air ventilation system in 10 hotels, 4 restaurants and 4 supermarkets was 86.0%.The over standard rate of PM₁₀, bacteria and fungi in air supply was 3.4%, 31.8% and 33.0%, respectively, but β-hemolytic streptococcus was not detected.The dust amount and microbial index on the inner surface of the air duct were up to standard.Another two indexes of fresh air quantity and Legionella pneumophila were tested in the hotel.The qualified rate of fresh air quantity was 25.6%, the positive rate of Legionella pneumophila type 1(LP1) in cooling water was 30.0%, and no Legionella pneumophila was detected in condensate water.The correlation analysis of 9 indexes showed that there was a significant positive correlation between PM₁₀ in air supply and in indoor air; the amount of microbial pollution on the inner surface of air duct and in air supply; and the amount of microbial pollution in air supply and in indoor air. Conclusions In Putuo District, the general sanitary condition of the centralized air ventilation system in public places is not to be optimistic.The qualified rate of bacteria and fungi in the air supply and fresh air quantity is low, and Legionella pneumophila was detected.The indoor air quality can be improved by installing efficient air purification device and regular cleaning and disinfection.

OBJECTIVETo construct DNA methyltransferase 1 (DNMT1) low expression 16HBE cell line and observe the variation of cell cycle and global genomic DNA methylation.

METHODSThe method of Lenti-virus induced RNA interference was applied to introduce four different shRNA fragment into 16HBE cells. Flow cytometry and 5-mC immunofluorescence methods were used to observe the cell cycle and global DNA methylation status of DNMT1 low expression 16HBE cells.

RESULTSThe DNMT1 protein relative expression level of 16HBE-shDNMT1-4 cell line was down regulated about 44% (P < 0.05) compared with the control. No obvious differences of cell cycle and global genome DNA methylation status were observed between the 16HBE and 16HBE-shDNMT1.

CONCLUSIONThe DNMT1 gene low expression cell is successfully constructed, and there are no obvious changes happened on the cell cycle and global genomic DNA methylation.

Cell Cycle ; Cell Line ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; metabolism ; DNA Methylation ; Down-Regulation ; Epithelial Cells ; metabolism ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics

OBJECTIVEThe aim of this study was to evaluate the effects of XRCCl gene polymorphisms and its haplotype on the susceptibility of pancreatic carcinoma.

METHODSPeripheral blood DNA was extracted from 210 pancreatic carcinoma patients and 213 control subjects. SNaPshot technique was used for genotyping seven SNP sites of the XRCCl gene (rs3213403, rs25487, rs1799782, rs731420, rs1001581, rs12611088, and rs3213282). Logistic regression model was performed to analyze the relationship of different genotypes or haplotype and the susceptibility of pancreatic carcinoma.

RESULTSThe frequency for allele A at site rs25487 in the case group was significantly higher than that in the control group (P < 0.05). The frequency of GG, GA and AA genotype between the case group and control group had statistically significant differences (P < 0.05). Compared with GG genotype, the risk of pancreatic carcinoma in the subjects carrying mutated allele A (GA+AA) was increased by 0.648 times (P < 0.05). Among them the pancreatic carcinoma risk of individuals carrying A allele was increased by 0.552 times compared with the individuals carrying G allele. The frequency of allele and genotype at site rs1799782 in the case group and control group had a significant difference (P < 0.05). Compared with the CC genotype, the risk of pancreatic carcinoma in the subjects carrying mutated allele T (CT+TT) was increased by 0.683 times. Among them the pancreatic carcinoma risk of individuals carrying T allele was increased by 0.549 times compared with the individuals carrying C allele. Significant differences were observed in linkage disequilibrium between any two of the seven SNPs (P < 0.05), the frequency of H4-AGCCCGC, H6-GGCCCGG or H7-AGCCTAG haplotypes was significantly lower in the case group than that in the control group (P < 0.05).

CONCLUSIONSThe single nucleotide polymorphisms of rs25487 and rs1799782 for XRCC1 gene may be correlated with the occurrence of pancreatic carcinoma. The haplotypes of H4-AGCCCGC, H6-GGCCCGG and H7-AGCCTAG might be a potential genetic protective factor for the occurrence of pancreatic carcinoma.

Alleles ; DNA-Binding Proteins ; genetics ; metabolism ; Genetic Predisposition to Disease ; epidemiology ; Genotype ; Haplotypes ; Humans ; Pancreatic Neoplasms ; epidemiology ; Polymorphism, Single Nucleotide ; X-Rays ; X-ray Repair Cross Complementing Protein 1

OBJECTIVETo investigate the correlation of serum sex hormones and parathyroid hormone (PTH) with the biochemical markers of bone turnover in aged men.

METHODSWe collected the laboratory data of 465 men aged 60- 93 (73. 1 +/- 8. 3) years old, who came for routine physical examinations in our hospital. We obtained the levels of serum follicle- stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), testosterone (T), sex hormone binding globulin (SHBG), PTH, 25-hydroxy-vitamin D3 (25(OH) D3), and bone turnover markers C-terminal telopeptide of type I collagen (CTX), osteocalcin (OC) and amino-terminal propeptide of type I procollagen (PINP). We also determined free testosterone (FT) , bioactive testosterone (BT) , testosterone secretion index (TSI) and FT index (FTI), and analyzed the correlation of each index with the biochemical markers of bone turnover.

RESULTSThe concentrations of serum FSH, LH, and SHBG increased, while the levels of FT, BT, TSI, FTI, PTH, CTX, OC and PINP decreased with age, especially in those over 80 years old (P <0.05). PTH was positively correlated with CTX, OC and PINP (r =0. 227, 0. 269 and 0. 162, P <0. 01), even after the adjustment for age, while SHBG negatively correlated with OC (r = -0. 100, P <0.05). The bone turnover markers increased with the elevation of the PTH quartiles, with significant differences between the first and the fourth quartile (P <0. 01). Multiple stepwise regression analysis showed that age was correlated inversely with CTX, OC and PINP ( beta = -0. 126, -0. 141 and -0. 122, P <0.05) , PTH positively with the three markers (beta = 0. 196, 0.279 and 0.189; P <0. 001), and SHBG negatively with OC ( beta = -0. 100, P <0.05) .

CONCLUSIONAging is the fundamental cause of reduced bone turnover in aged men. The levels serum PTH and SHBG are significantly associated with the biochemical markers of bone turnover.

Aged ; Aged, 80 and over ; Aging ; Bone Density ; Bone Remodeling ; physiology ; Bone and Bones ; metabolism ; Estradiol ; blood ; Follicle Stimulating Hormone ; blood ; Gonadal Steroid Hormones ; blood ; Humans ; Male ; Middle Aged ; Parathyroid Hormone ; blood ; Testosterone ; blood