[Reevaluation of equivocal HER2 status detected by immunohistochemistry according to the 2019 guidelines for HER2 detection].

Objective To understand the effects and clinical significance of the 2019 guidelines for the human epidermal growth factor receptor 2 (HER2) detection. Methods: According to the 2014 guidelines, 548 cases of invasive breast cancer with equivocal HER2 (2+) detected by immunohistochemistry (IHC) in Taizhou Enze Medical Center, Zhejiang Province, China from 2013 to 2019 were selected. The results of IHC and HER2/CEPl7 double-probe were reevaluated and divided into groups according to the 2019 guidelines for the comparative analysis. Results: Among the 548 IHC HER2 (2+) invasive breast cancers, the number of positive, equivocal and negative cases for HER2 were 96 (17.52%), 81 (14.78%) and 371 (67.70%), respectively, according to the 2014 guidelines. However, according to the 2019 guidelines, 10 cases (1.82%) were reclassified as IHC 1+, 2 cases in the group 2 were reclassified as negative, and all the originally equivocal cases in group 4 were reclassified as negative. Finally, the total number of positive and negative cases for HER2 were 94 (17.15%) and 454 (82.85%), respectively. Conclusions: After applying the 2019 guidelines, the number of IHC 2+ cases decreases, and the positive rate for HER2 also decreases slightly due to the reevaluation change in groups 2 and 4, leading to reclassification of the cases that were deemed equivocal according to the 2014 guidelines. In general, the new 2019 guidelines are more reasonable and easier to use.

Development and characterization of Rift Valley fever virus-like particles.

Rift Valley fever (RVF) is an acute, febrile zoonotic disease that is caused by the RVF virus (RVFV) and spread by arthropod vectors. RVF is currently prevalent in Africa and the Arabian Peninsula, and causes substantial economic losses. Furthermore, this disease poses a serious threat to animal and human health in regions worldwide, making it a serious public health concern. However, RVFV vaccines for human use are still unavailable, and hence there is an urgent need for novel efficient vaccines against RVFV. Vaccine preparation techniques have become a crucial factor in developing new vaccines. In the current study, the N and G protein genes of RVFV were inserted into the pFastBacDual baculovirus expression vector downstream of the pP10 and pPH promoters. The resultant recombinant vector, pFastBacDual-S-M, was transfected into Sf9 insect cells by lipofection. The recombinant baculovirus, named rBac-N-G, was retrieved and infected into Sf9 insect cells to generate RVFV virus-like particles (VLPs). Using polyclonal antibodies against RVFV proteins in immunofluorescence and western blot analyses, we positively identified the presence of the RVFV proteins in VLP preparations. Sucrose density gradient centrifugation and transmission electron microscopy revealed that the morphology of the RVFV VLPs was consistent with previous reports of RVFV virions. This study describes a technique for efficient production of RVFV VLPs, and has laid the foundation for future VLP-based RVFV vaccines.

A prospective analysis of the effects of enteric-coated mycophenolate sodium and mycophenolate mofetil co-medicated with a proton pump inhibitor in kidney transplant recipients at a single institute in China.

BACKGROUND: Enteric-coated mycophenolate sodium (EC-MPS) and mycophenolate mofetil (MMF), two prodrugs of mycophenolic acid (MPA), have been used in immunosuppressive regimens. After being taken orally, both of them transform to MPA to achieve immune suppression effects; however, the main site of absorption and metabolism of EC-MPS is different from that of MMF in vivo. Therefore, combined application with related drugs may result in different MPA levels and have different clinical effects in kidney transplant recipients. OBJECTIVE: To evaluate the efficacy of EC-MPS compared with MMF in Chinese renal transplant patients comedicated with a proton pump inhibitor (PPI). METHODS: Our subjects were 88 patients who received renal transplants at the 309th Hospital of the Chinese PLA from May 2010 to April 2013. These were made up of two groups including 27 patients with EC-MPS and 61 with MMF. The immunosuppression regimen was EC-MPS/MMF + cyclosporine/tacrolimus + steroid hormone, comedicated with a PPI (omeprazole). The patients' levels of exposure of MPA within 1 week after operation were monitored. Clinical indicators such as incidence of delayed graft function and acute rejection, the rate of change of serum creatinine hemoglobin, leucocytes, and neutrophils, as well as clinical adverse drug reactions and drug conversion were analyzed retrospectively. RESULTS AND CONCLUSION: The kidney function of patients recovered to normal in both the EC-MPS and MMF groups. The mean concentration to peak (Cmax), the mean half-life (t1/2), and the area under the concentration-time curve (AUC0-12) of MPA in the EC-MPS group were higher than those in the MMF group (P < .05). This indicated that the pharmacokinetic parameters for MPA when EC-MPS is co-administered with a PPI in kidney transplant patients in China is better than for comedication with MMF and a PPI. The MMF group had a higher incidence of drug withdrawal because of higher infection rates, leucocyte decrease, and more gastrointestinal side effects than the EC-MPS group (P < .05). No significant differences in the clinical effect on kidney transplant recipients were observed in the limited observation time.

Clinical relevance of sHLA-G-mediated with better graft acceptance in early posttransplantation.

The aim of this study was to measure the level of soluble human leukocyte antigen (sHLA-G) in renal transplant patients, to determine the relationship between these levels and the occurrence of acute rejection episodes, and to identify their influence on graft acceptance early posttransplantation. sHLA-G, as measured by an enzyme-linked immunosorbent assay, was significantly increased (P < .01) early posttransplantation (3 months); the other group maintained low levels throughout the study. The latter group displayed a high incidence of acute rejection episodes and a lower clearance of serum creatinine with a longer period for hemoglobin to recover to normal (P < .01). These results suggested that HLA-G participates in the induction of immunologic tolerance in these recipients.

Regulation of adipocyte differentiation by PEGylated all-trans retinoic acid: reduced cytotoxicity and attenuated lipid accumulation.

Obesity is major risk factor for many disorders, including diabetes, hypertension and heart disease. Unfortunately, there is a dearth of therapeutic agents available to clinicians for the treatment of obesity. The principal aim of this study was to investigate whether PEGylated all-trans retinoic acid (PRA) can have favorable stability and biological activity in 3T3-L1 preadipocytes as an antiobesity drug. Here, we found that PRA inhibits the process of adipogenesis, including survival of adipocytes and differentiation to mature adipocytes. The results showed that RA nanoparticles (NPs) were prepared by PEGylation; below 200 nm, PRA-NPs were obtained. Moreover, PRA decreased glycerol-3-phosphate dehydrogenase activity in 3T3-L1 preadipocytes by acting with major adipocyte marker proteins such as PPARgamma2, C/EBPalpha and aP2 modulators. Apoptosis, in addition, increased as the level of RA increased from 10 to 20 microM, whereas PRA reduced apoptosis with increasing concentrations. Our data suggest that PRA-NP has potential as an antiobesity drug carrier due to its small particle size and PEGylated core-shell structure. In addition, our results suggest that PRA inhibits the process of adipogenesis and may be developed to treat obesity. Based on these results, PRA is suitable for adipocyte studies, and an enhanced effect of PRA with adipocyte differentiation offers a challenging approach for pharmaceutical applications.

[Observation of repair of wounded rat skin by affinity histochemical method of SJA].

Time expression of SJA tissue receptors surrounding wounded skin of 40 rats was studied by affinity histochemical method. The results were compared to that by EGFR immunohistochemical method and c-myc situ hydridization method. It was observed that SJA receptors began to decrease after the skin was injured and reached the lowest quantities in 15 minutes. After that, they began to increase and reached the highest quantities in 2 hours. It needs to be further studied whether there is the correlation between SJA receptors and EGFR has the function of repairing wounded skin.

[Inhibitory effect of methanol extract of Boschniakia rossica Fedtsch. et Flerov on rat hepatic preneoplastic lesions induced by diethylnitrosamine].

OBJECTIVE: To investigate the inhibitory effect of Boschniakia rossica(BR) on hepatocarcinogenesis in rats. METHOD: Based on immunohistochemistry techniques, the expression of placental form glutathione S-transferase(GST-P), mutant p53 and p21 protein were investigated in hepatic preneoplastic lesions induced by Solt-Farber protocol in the liver of rats that had been treated with the above method, administered with BR extract and of control group. RESULT: The extract of BR(500 mg/kg) has inhibitory effect on the formation of diethylnitrosamine-induced GST-P-positive foci in F344 rat and the expression of mutant p53 and p21 protein was lower than that of hepatic preneoplastic lesions, and the increasing gamma-glutamyltranspeptidase(gamma-GT) activity in rat liver treated with Solt-Farber protocol was decreased by the extract of BR. CONCLUSION: These results indicate that BR has inhibitory effect on DEN induced hepatic preneoplastic lesions in F344 rat.

Separation of carotenes on cyclodextrin-bonded phases.

The separation of carotenoids and retinoids on a beta-cyclodextrin-bonded stationary phase with conventional mobile phases is reported. Compounds studied include beta-carotene (all-trans), 15,15'-cis-beta-carotene, 7,8,7',8'-dihydro-beta-carotene, alpha-carotene, lycopene, lutein, zeaxanthin, retinal, retinol, retinol palmitate and retinol acetate. The best resolution of carotenes was obtained with low concentrations (less than or equal to 1%) of polar solvents (e.g., 2-propanol or ethyl acetate) in hexane or cyclohexane. Xanthophylls required much higher concentrations of polar solvents. The best solvent for the resolution of lutein and zeaxanthin was found to be dichloromethane. The resolution of cis/trans-isomers and the tentative identification of other isomers present in newly synthesized carotenoid standards is also reported. All-trans-isomers were found to be eluted before cis-isomers.

Cyclodextrin chiral stationary phases for liquid chromatographic separations of drug stereoisomers.

Many active drugs are racemic mixtures. Because the two enantiomers of a racemate often cause different pharmacological responses, the use of optically pure isomers is desirable and may be soon required. Cyclodextrin-bonded silica gel can be used as chiral stationary phase (CSP) in liquid chromatography. The enantiomers of 25 different racemic drugs were separated on such CSPs in the reversed-phase mode. The principal features of the cyclodextrin chiral recognition mechanism are recalled and some information on future trends for cyclodextrin CSPs is provided.

Liquid chromatographic separation of anomeric forms of saccharides with cyclodextrin bonded phases.

A brief review of sugar stereochemistry is given. The separation of 34 different pairs of anomers was accomplished on both alpha- and beta-cyclodextrin columns. Five different mobile phases were evaluated. The separation of anomers could be enhanced or suppressed by altering the mobile-phase composition, column temperature, flow rate, and so on. The separation of anomers that mutarotate is somewhat more difficult than those that do not. Prior knowledge as to the rate of mutarotation is useful so that the chromatographic conditions can be arranged to minimize any deleterious effects on the separation.

Optical enrichment of dansyl-rac-amino acids by formation of crystalline inclusion complexes with cyclodextrins.

Optical enrichment from racemic dansyl-leucine, dansyl-norleucine, and dansyl-phenylalanine with both beta- and gamma-cyclodextrins in water is reported. Initial crystallization yielded the dansyl-L-Leucine isomer complexed in excess with beta-cyclodextrin with an optical purity of 62-78% depending on experimental conditions. The optical purities obtained for L-norleucine and L-phenylalanine were 71 and 64%, respectively. The optical purity can be increased with continued recrystallization. The dansyl-D-leucine isomer was obtained in the mother liquor with an optical purity of 54-93% depending on experimental conditions. The optical purities obtained for D-norleucine and D-phenylalanine were 72 and 58%. The optical purity of the isomer depended on the molar ratio of host:guest and the pH value of the solution. Optimum enrichment of both enantiomers was achieved with host:guest ratios of 2:1 and 3:1. Although maximum crystalline yield of the dansyl-leucine/CD inclusion complex was obtained at a pH of 3.5, optical purity of both enantiomers was less than that obtained at other pHs. The influence of the molar ratio of host:guest and the pH value of the solution are discussed. This method is suitable for large-scale enantiomeric separations.

Evaluation of the liquid chromatographic separation of monosaccharides, disaccharides, trisaccharides, tetrasaccharides, deoxysaccharides and sugar alcohols with stable cyclodextrin bonded phase columns.

Both alpha- and beta-cyclodextrin (CD) bonded phase columns were evaluated for their ability to separate carbohydrates and related molecules. Chromatographic data on approximately 50 solutes are reported. Mobile phases consisting of acetonitrile-water or acetone-water produce the best separations. Separations could be run in isocratic or gradient modes. The alpha-CD column produced slightly more efficient separations than the beta-CD column. Both cyclodextrin columns seemed to be more efficient and selective than alkylamine and ion-exchange columns. The stability and reproducibility of the CD columns were excellent and showed little deterioration after several thousand injections. The retention of saccharides on CD bonded phases seems to be related to the number of available hydroxy groups per solute and to the size of the solute. Analysis times and detection sensitivity are discussed.