ABSTRACT
Cryptorchidism is associated with infertility in adulthood. Early orchiopexy is suggested to reduce the risk. Information is lacking on the potential link between infant germ cell maturation and the risk of future infertility. The objective of the study was to evaluate age-related germ cell development in cryptorchidism. Immunostaining for markers of germ cell development (octamer-binding transcription factor 3/4 [OCT3/4], placental alkaline phosphatase [PLAP], KIT proto-oncogene [C-KIT], podoplanin [D2-40], Lin-28 homolog A [LIN28], and G antigen 7 [GAGE-7]) was performed in testicular biopsies from 40 cryptorchid boys aged 4-35 months. Germ cell numbers and distributions were evaluated in cross sections of seminiferous tubules, with and without immunostaining. OCT3/4, D2-40, and LIN28 were generally expressed in the early stages of germ cell development, as shown by positive expression in germ cells in the central region of seminiferous tubules. In contrast, PLAP and GAGE-7 were expressed in both central and peripheral parts of the tubules in the early stages of development and expressed mainly in a peripheral position with advancing age. Germ cell maturation was delayed in this study population as compared with that observed in our previous study on germ cell markers in a healthy population. The number of GAGE-7-positive germ cells per tubular cross section obtained by immunostaining was significantly higher than that obtained by standard hematoxylin and eosin staining. Double immunostaining revealed heterogeneity in germ cell development in cryptorchid testes. These results shed light on the pathophysiology of germ cell development in boys with cryptorchidism.
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AIM:To observe the the effect of vitreomacular adhesion on the anti-vascular endothelial growth factor (VEGF) treatment in patients with macular edema (ME) induced by branch retinal vein occlusion (BRVO).METHODS:A retrospective clinical study.Totally 58 patients(60 eyes) were induced in this study during April 2014 to May 2016,who were diagnosed with macular edema due to branch retinal vein occlusion.All subjects were assigned to 2 groups according to the examination of optical coherence tomography (OCT):combined with vitreomacular adhesion groups (Group A) and uncombined with vitreomacular adhesion groups (Group B).The basic situation of the two groups was as follows:Group A:23 cases (24 eyes),the average age was 55.91± 7.34,the mean disease course was 3.4±1.01mo,the mean macular central retinal thickness (CMT) was 463.26±53.73μm and the average BCVA was 0.63±0.11;Group B:35 cases (36 eyes),the average age was 56.33± 5.34,the mean disease course was 2.82± 1.33mo,the mean CMT was 482.90± 37.43μ m and the average BCVA was0.59±0.12.All cases received vitreous injection of 0.5mg Conbercept.Injections were repeated based on the visual changes and the OCT findings.The follow-up time was more than 6mo.BCVA,CMT,and the numbers of injections of two groups were recorded at pre-operation and postoperative 1,6mo and the statistical analysis was conducted.RESULTS:BCVA and CMT of the most patients were improved compared to prior treatment and the difference had statistical significance (P ＜ 0.05).There were significant differences on CMT between two groups at 1 and 6mo after treatment (t=9.13,10.01;P＜0.05).While BCVA between two groups at 1 and 6mo after treatment was not statistically different (t=2.13,5.32;P＞0.05).At 6mo after treatment,the average numbers of injections in combined vitreomacular adhesion groups were 4.38±0.97 times and the average numbers of injections in uncombined vitreomacular adhesion groups were 3.56 ± 0.71 times.The difference had statistical significance (t=4.56,P＜ 0.05).Systematic adverse reactions and persistent intraocular pressure elevation,retinal detachment,endophthalmitis,vitreous hemorrhage were never found in the follow-up period.CONCLUSION:Vitreous injection of anti-VEGF treatment for macular edema induced by branch retinal vein occlusion has good clinical results.However,if there is merged with vitreomacular adhesion at the same time,the treatment effect of anti-VEGF will be weakened to some extent.
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The enzymatic browning is one of the main reasons for affecting the quality of medicinal flowers. In the process of chrysanthemum harvesting and processing, improper treatment will lead to the browning and severely impact the appearance and quality of chrysanthemum. Peroxidase enzyme is one of the oxidoreductases that cause enzymatic browning of fresh chrysanthemum. The enzymatic characteristics of peroxidase (POD) in chrysanthemum were studied in this paper. In this experiment, the effects of different reaction substrates and their concentrations, PH value of buffer and reaction temperatures on the activity of POD enzyme were investigated. The results showed that the optimal substrate of POD was guaiacol, and the optimal concentration of POD was 50 mmol·L⁻¹. The optimal pH value and reaction temperature were 4.4 and 30-35 °C, respectively. Michaelis-Menten equation was obtained to express the kinetics of enzyme-catalyzed reaction of POD, Km=0.193 mol·L⁻¹, Vmax=0.329 D·min⁻¹. In addition, the results of POD enzyme thermal stability test showed that the POD enzyme activity was inhibited when being treated at 80 °C for 4 min or at 100 °C for 2 min. The above results were of practical significance to reveal the enzymatic browning mechanism, control the enzymatic browning and improve the quality of chrysanthemum, and can also provide the basis for the harvesting and processing of medicinal materials containing polyphenols.
ABSTRACT
To investigate the differences of chemical compositions in Gynostemma pentaphyllum leaves prepared by different processing methods. Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to compare the chemical compositions between shade-dried processing and drum-dried processing. Forty six gypenosides were identified by control comparison， liquid chromatography-mass spectrometry(LC-MSn) fragmentation information， and literature data. The mass spectral peak area statistics was combined with principal component analysis(PCA)， and the results showed that eight batches of Gynostemma pentaphyllum leaves samples were divided into two groups according to the two different processing methods; ten chemical compositions with significant differences were screened according to mass spectrum information combined with partial least-squares discriminant analysis(PLS-DA). The result showed that most parent nucleus of the gypenosides contained three to four glycosides in drum-dried samples， and one to two glycosides in the shade-dried samples. It was inferred from further MS analysis that desugarization of gypenosides was present to produce secondary glycosides with the effect of glucosidase in the shade-drying， thus resulting in difference in compositions. This study provided data support for harvesting， processing and quality control of Gynostemma pentaphyllum leaves.
Subject(s)
Chromatography, High Pressure Liquid , Gynostemma , Chemistry , Mass Spectrometry , Plant Leaves , Chemistry , Saponins , ChemistryABSTRACT
Seven compounds(deacetylasperulasidic acid methyl ester, gardenoside, chlorogenic acid, geniposide, crocin-Ⅰ, crocin-Ⅱ, chikusetsu saponin Ⅳa)were determined simultaneously by multiple wavelength HPLC with diode array detector(DAD) in different parts of Gardenia jasminoides. The results showed that these components in different parts of G. jasminoides had a different distribution, and there was a large difference in content of each component. Geniposide was mainly distributed in fruits and leaves; chikusetsu saponin Ⅳa was mainly distributed in roots and stems; crocus glycosides existed mainly in fruits; chlorogenic acid had a higher distribution in leaves and stems; gardenoside had a higher distribution in leaves and roots, while ceacetylasperulasidic acid methyl ester had a higher distribution in roots and stems. Based on the analysis of the chemical composition and content difference in different parts of G. jasminoides, the basis for the comprehensive utilization and quality evaluation of resources of G. jasminoides was provided.
ABSTRACT
To optimize the purification process of gynostemma pentaphyllum saponins (GPS) based on "adjoint marker" online control technology with GPS as the testing index. UPLC-QTOF-MS technology was used for qualitative analysis. "Adjoint marker" online control results showed that the end point of load sample was that the UV absorbance of effluent liquid was equal to half of that of load sample solution, and the absorbance was basically stable when the end point was stable. In UPLC-QTOF-MS qualitative analysis, 16 saponins were identified from GPS, including 13 known gynostemma saponins and 3 new saponins. This optimized method was proved to be simple, scientific, reasonable, easy for online determination, real-time record, and can be better applied to the mass production and automation of production. The results of qualitative analysis indicated that the "adjoint marker" online control technology can well retain main efficacy components of medicinal materials, and provide analysis tools for the process control and quality traceability.
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<p><b>OBJECTIVE</b>To explore the clinical efficacy and safety of EPOCH±R followed by DICE±R regimen for primary breast diffuse large B-cell lymphoma.</p><p><b>METHODS</b>Forty-three patients with primary breast diffuse large B-cell lymphoma were admitted in our hosptial from January 2000 to April 2016. Among them 24 patients were treated with CHOP±R regimen, 19 patients were treated with EPOCH±R followed by DICE±R regimen. The clinical efficacy, survival rate and adverse effects were observed and compared between them.</p><p><b>RESULTS</b>The complete rate in EPOCH±R followed by DICE±R regimen group was higher than that in the CHOP±R group (84.2% vs 70.8%), and the relapsed rate was lower in EPOCH±R followed by DICE±R regimen group than that in the CHOP±R group (6.25% vs 35.3%). Progression-free survival (PFS) and overall survival (OS) rates of 5 years after diagnosis in the EPOCH±R followed by DICE±R group were significantly higher as compared with that in CHOP±R group (PFS, 75% vs 47.4%, P=0.035; OS, 73.3% vs 45.2%, P= 0.043). Treatment-related hematologic adverse events were more serious in the EPOCH±R followed by DICE±R group(63.2% vs 25%). However, these adverse events were controlled and no treatment-related deaths were observed. Multivariate analysis showed that age (P=0.008; 95% CI, 0.026 to 0.579), radiotherapy (P=0.045; 95% CI, 1.028 to 14.719) and LDH level (P=0.007; 95% CI, 0.017 to 0.531) were independent prognostic factors for 5 year overall survival.</p><p><b>CONCLUSION</b>EPOCH±R followed by DICE±R regimen is an effective and safe treatment regimen for PB-DLBCL. Prognostic factors for survival are age, LDH level and radiotherapy.</p>
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AIM: To observe the effect of perioperative detail treatment of pterygium. METHODS:Sixty cases(100 eyes) of pterygium patients were collected from Sep. 2012 to Jan. 2014. All cases were performed modified pterygium excision with limbal epithelial autograft transplantion under microscope. Corneal wound healing, graft survival, and the recurrence of pterygium were observed. RESULTS:All cases of conjunctival flap survive, corneal wound closed well. The patients were followed up for 7 to 12mo. Among them, 3 eyes recurred and the recurrence rate was 3%. There were no serious complications. CONCLUSION:In the operation of pterygium excision with limbal epithelial autograft transplantion, standard and detail treatment can significantly improve the clinical effect.
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The aim of this study was to illustrate the mechanism of inhibiting the cell growth in diffuse large B-cell lymphoma by histone deacetylase inhibitor valproic acid (VPA) combined with mTOR inhibitor temsirolimus (TEM). MTT assay and Wright's stain were used to assess cell growth inhibition and to detect the cell morphological changes respectively. The cell apoptosis, cell cycle and cell autophagy were determined by flow cytometry. Ultrastructure changes were confirmed by electron microscopy. Protein changes were detected by Western blot. The results showed that both VPA and TEM alone inhibited cell proliferation and the effect was more obvious in the combination group. VPA combined with TEM induced cell arrest in G0/G1 phase and upregulated the expression of autophagy-related protein LC3, without cell apoptosis. Moreover, typical autophagosomes were observed, further confirming the presence of autophagy. Western blot showed the changes of proteins involved in autophagy signaling pathway. VPA decreased HDAC1 and HDAC3 expression and increased histone acetylation, suggesting that VPA also affected lymphoma cell proliferation through epigenetic modification. It is concluded that the combined treatment of VPA and TEM induces cell cycle arrest and cell autophagy, which provides a new clue for their clinical application in diffuse large B-cell lymphoma.
Subject(s)
Humans , Autophagy , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , Epigenesis, Genetic , Histone Deacetylase 1 , Metabolism , Histone Deacetylases , Metabolism , Lymphoma, Large B-Cell, Diffuse , Pathology , Microtubule-Associated Proteins , Metabolism , Sirolimus , Pharmacology , Valproic Acid , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of epidermal fatty acid-binding protein (E-FABP) and fatty acid synthase (FAS) in human breast cancer and identify the potential markers and therapeutic targets for breast cancer.</p><p><b>METHODS</b>FAS and E-FABP expressions were detected in 76 patients with infiltrating ductal breast carcinoma using RT-PCR, immunohistochemistry and Western blotting. The possible associations of the expression of the two proteins with the major clinicopathological factors were analyzed.</p><p><b>RESULTS</b>E-FABP and FAS expression levels were significantly decreased (P<0.05) in grade III as compared with grades I and II infiltrating ductal breast carcinoma. There was a positive correlation between E-FABP and FAS expressions, but their expressions were not correlated to the clinicopathological factors of the patients except for the tumor grades. High E-FABP expression level in grades I and II tumors were associated with an early increased responsiveness to FAS.</p><p><b>CONCLUSION</b>The variation of the E-FABP and FAS expressions in the lesions is associated with increase of the risk for breast cancer, and the results of this study provide evidence for developing new molecular markers of high-risk lesions and identifying new the targets for breast cancer therapy.</p>
Subject(s)
Adult , Aged , Female , Humans , Middle Aged , Biomarkers, Tumor , Genetics , Blotting, Western , Breast Neoplasms , Genetics , Metabolism , Pathology , Carcinoma, Ductal, Breast , Genetics , Metabolism , Pathology , Fatty Acid Synthases , Genetics , Fatty Acid-Binding Proteins , Genetics , Immunohistochemistry , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of signal factors of corticosterone (CS), cAMP, cGMP, Ca2+ andprotein kinase C (PKC) on lymphocyte apoptosis in mouse thymus induced by X-rays of 4 Gy in vitro.</p><p><b>METHODS</b>The DNA lytic rate for thymocytes was measured by fluorospectrophotometry.</p><p><b>RESULTS</b>The DNA lyric rate for thymocytes 4-8 hours after irradiation with 2-8 Gy was significantly higher than that in the control (P<0.01). As compared with the control, the DNA lytic rate for thymocytes treated with 0.01 micromol/L CS (P<0.01), 50 ng/mL cAMP (P<0.01), 0.05-0.4 microg/mL ionomycin (Iono, P<0.05 or P<0.01) or 0.05-0.4 ng/mL phorbol myristate acetate (PMA, P<0.05 or P<0.01), respectively, was significantly increased, while the rate for thymocytes treated with 50 ng/mL cGMP was not significantly increased. The DNA lytic rate for thymocytes treated with 0.01 micromol/L CS (P<0.01), 50 ng/mL cAMP (P<0.01), 0.2 and 0.4 microg/mL Iono (P<0.05), and 0.2 and 0.4 ng/mL PMA (P<0.05) plus 4-Gy irradiation, respectively, was significantly higher than that treated with single 4-Gy irradiation, while the rate for thymocytes treated with 50 ng/mL cGMP plus 4-Gy irradiation was not increased. When both 0.4 microg/mL Iono and 0.4 ng/mL PMA acted on the thymocytes, the DNA lytic rate for thymocytes was significantly higher than that in the control (P<0.01), the DNA lytic rate for thymocytes treated with both 0.4 microg/mL Iono and 0.4 ng/mL PMA plus 4-Gy irradiation was significantly higher than that treated with single 4-Gy irradiation (P<0.05), but was not significantly higher than that treated with 0.4 microg/mL Iono plus 4-Gy irradiation or 0.4 ng/mL PMA plus 4-Gy irradiation.</p><p><b>CONCLUSION</b>CS, cAMP, Ca2+, and PKC signal factors can promote thymocyte apoptosis induced by larger dose X-rays.</p>
Subject(s)
Animals , Male , Mice , Apoptosis , Radiation Effects , Calcium , Pharmacology , Corticosterone , Pharmacology , Cyclic AMP , Pharmacology , Cyclic GMP , Pharmacology , Ionomycin , Pharmacology , Protein Kinase C , Metabolism , Spectrometry, Fluorescence , Tetradecanoylphorbol Acetate , Pharmacology , Thymus Gland , Cell Biology , X-RaysABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of liver-type fatty acid-binding protein (L-FABP) and vascular endothelial growth factor (VEGF) in human hepatocellular carcinoma (HCC) and its adjacent liver tissues, and investigate the correlation between the expressions of L-FABP and VEGF and their role in the occurrence and progression of HCC.</p><p><b>METHODS</b>Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical (IHC) staining were employed to examine the expression of L-FABP and VEGF in HCC and its adjacent liver tissues obtained from the surgical specimens of 61 HCC patients who underwent liver resections in West China hospital.</p><p><b>RESULTS</b>The results of RT-PCR showed that the expression level of L-FABP and VEGF in HCC was significantly higher than that in its adjacent liver tissues (L-FABP: 0.97-/+0.12, 0.83-/+0.14, t=5.21, P<0.05; VEGF: 0.92-/+0.11, 0.59-/+0.15, t=11.79, P<0.05). L-FABP tended to co-express with VEGF (P<0.05). IHC staining revealed that the expression of L-FABP and VEGF was mainly located in the cytoplasm, and the gray scale of L-FABP expression was significantly higher than that in the adjacent liver tissues (92.73-/+7.67, 82.83-/+6.90, t=7.44, P<0.05). The number of L-FABP- and VEGF-positive cells in HCC was significantly lower than that in the adjacent liver tissues (L-FABP: 92.18-/+4.44, 84.52-/+6.43, t=5.94, P<0.05; VEGF: 88.69-/+5.56, 77.64-/+5.93, t=8.72, P<0.05). Co-expression of L-FABP and VEGF observed in RT-PCR and also in IHC (P<0.05).</p><p><b>CONCLUSION</b>Both L-FABP and VEGF expressions are up-regulated in HCC. L-FABP gene may be involved in the carcinogenesis of human HCC. Expression of L-FABP is associated with VEGF expression, suggesting that L-FABP promotes the growth of blood vessels by taking up the fatty acids from the bloodstream, and both of them produce a marked effect on energy metabolism in HCC.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Blotting, Western , Carcinoma, Hepatocellular , Genetics , Metabolism , Pathology , Fatty Acid-Binding Proteins , Genetics , Gene Expression Regulation, Neoplastic , Immunohistochemistry , Liver Neoplasms , Genetics , Metabolism , Pathology , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Vascular Endothelial Growth Factor A , GeneticsABSTRACT
OBJECTIVE@#To investigate the clinical significance of the changes of vascular endothelial growth factor ( VEGF ) and interleukin-6 ( IL-6 ) level in multiple myeloma (MM), solid tumor following bone metastasis.@*METHODS@#Thirty- seven MM patients, including 7 in Stage I , 8 in Stage II , 22 in Stage III, 8 solid tumor with bone metastasis patientsly, and 17 healthy controls were enrolled in this study. The serum VEGF and IL-6 levels were determined by ELISA.@*RESULTS@#Serum VEGF and IL-6 concentrations in patients with MM and solid tumor were significantly higher than those of the healthy controls (P <0.01 ), and the VEGF level was higher in MM than in solid tumor with bone metastasis. There was significant difference in VEGF and IL-6 levels in various clinical stages of MM. VEGF levels in Stage II were significantly higher than in Stage I (P < 0.05 ) and IL-6 levels in Stage II were significantly higher than in Stage I (P < 0.05). The levels of IL-6 showed great difference according to bone lesion scores (P < 0.05). There was a positive correlation between IL-6 and serum calcium or C-reactive protein( P <0.01) , and there was a positive correlation between VEGF and serum Cr or urinary Bene-Jones protein lambda (P < 0.01 ). The IL-6 levels had significant differences between patients with the normal serum CRP, serum calcium, and beta2-MG and patients with abnormal ones (P < 0.05). VEGF levels showed significant differences between the patients with normal serum Cr, serum calcium Bene-Jones protein lambda, and urinary Bene-Jones protein lambda and patients with abnormal ones (P < 0.05).@*CONCLUSION@#Serum VEGF and IL-6 levels are helpful to diagnose the clinical stages, and understand bone lesion and serevity of MM.