[Establishment of mouse lens epithelial cell lines with the genotype of Hsf4-/-].

OBJECTIVE: To establish mouse lens epithelial cell lines with the genotype of Hsf4-/-. METHODS: The expended mouse lens epithelial cells, which were generated from P6 Hsf4-deficient mouse lens epithelia, were immortalized with SV40-T-antigen and named MLEC/Hsf4-/- cell. The expression of alpha A-crystallin was immunoblotted. Hsf4b cDNA was reconstituted by transiently transfection. RESULTS: The SV40-immortalized cells were in adherent growth mode with spindle morphology, pseudopodia, clear nuclear boundary membrane and cytoplasm translucent. Immunoblotting results indicated that the lens biomarker protein alpha A-crystallin was expressed in MLEC/Hsf4-/- cells. Reconstitution of Hsf4b into MLEC/Hsf4-/- cells upregulated the expression of Hsp25. CONCLUSIONS: The SV40-immortalized MLEC/Hsf4-/- cells have the lens epithelial characteristics and could be used as a tool for studying the signal transduction in vitro.

Tyrosine metabolic reprogramming coordinated with the tricarboxylic acid cycle to drive glioma immune evasion by regulating PD-L1 expression.

Due to the existence of the blood-brain barrier in glioma, traditional drug therapy has a poor therapeutic outcome. Emerging immunotherapy has been shown to have satisfactory therapeutic effects in solid tumors, and it is clinically instructive to explore the possibility of immunotherapy in glioma. We performed a retrospective analysis of RNA-seq data and clinical information in 1027 glioma patients, utilizing machine learning to explore the relationship between tyrosine metabolizing enzymes and clinical characteristics. In addition, we also assessed the role of tyrosine metabolizing enzymes in the immune microenvironment including immune infiltration and immune evasion. Highly expressed tyrosine metabolizing enzymes 4-hydroxyphenylpyruvate dioxygenase, homogentisate 1,2-dioxygenase, and fumarylacetoacetate hydrolase not only promote the malignant phenotype of glioma but are also closely related to poor prognosis. The expression of tyrosine metabolizing enzymes could distinguish the malignancy degree of glioma. More importantly, tyrosine metabolizing enzymes regulate the adaptive immune process in glioma. Mechanistically, multiple metabolic enzymes remodel fumarate metabolism, promote α-ketoglutarate production, induce programmed death-ligand 1 expression, and help glioma evade immune surveillance. Our data suggest that the metabolic subclass driven by tyrosine metabolism provides promising targets for the immunotherapy of glioma.

A comparative study on ultrasound-guided elite, Mammotome, and core needle biopsy for diagnosing malignant breast masses.

Introduction: The present study aims to clarify the advantages and disadvantages of elite biopsy, to provide a reference for selecting the puncture method. Material and methods: A total of 802 patients with a BI-RADS grade ≥ 4, as evaluated by the molybdenum target, and measurable lesions revealed by colour Doppler ultrasound, who were admitted at our department from January 2017 to January 2018, were enrolled in the present study. These patients were randomly divided into three groups: elite, Mammotome and core needle biopsy groups. The pathological underestimation rate, diagnostic accordance rate, haematoma incidence rate, and costs of these three biopsy methods were compared. Results: The difference in diagnostic accordance rates between the elite biopsy group and core needle biopsy group was statistically significant (98.9% vs. 94.7%, p = 0.003), as well as between the Mammotome biopsy group and core needle biopsy group (99.6% vs. 94.7%, p < 0.001). The difference in pathological underestimation rates between the elite biopsy group and core needle biopsy group was statistically significant (7.2% vs. 37.3%, p < 0.001), as well as between the Mammotome biopsy group and core needle biopsy group (1.6% vs. 7.2%, p < 0.001). The difference between the Mammotome biopsy group and elite biopsy group was not statistically significant. The incidence of haematoma in the Mommotome, elite, and core needle groups was 15.9%, 13%, and 21.7%, respectively (13% vs. 21.7%, p = 0.021). Conclusions: Elite biopsy has a low rate of pathological underestimation and low incidence of haematoma, can improve the breast conserving rate, and has an affordable cost. As a biopsy method with high accuracy, safety, and economy, elite biopsy can be widely used in clinics.

Experimental study on the clinical effects of Xiaoru Sanjie Jiaonang on mammary glands hyperplasia and ki-67.

OBJECTIVE: This study aims to observe the effect and mechanism of Xiaoru Sanjie Jiaonang (XRSJ) on the treatment of mammary gland hyperplasia, and provide a theoretical basis and clinical evidence for clinical expansion. METHODS: Japanese white rabbits were randomly divided into three groups: high-, middle- and low-dose groups; Xiaoyao Pill group; model control group; normal control group. The observation points were as follows: before XRSJ administration, three months after XRSJ administration, and three months after XRSJ discontinuance. Changes in breast height, morphological changes of the mammary gland under a light and electron microscope, and the expression of ki-67 were observed. At the same time, patients diagnosed with mammary gland hyperplasia at an Outpatient Clinic were selected and divided into treatment groups. These patients received XRSJ and Xiaoyao Pills, respectively, for one month, while patients in the control group did not receive any drug treatment. Clinical efficacy was observed while rechecking at the Outpatient Clinic after three months. Treatment with a therapeutic dose of XRSJ could significantly reduce breast height, decrease the number of lobules and acini in hyperplastic mammary glands and the layer number of ductal glandular epithelial cells, substantially lower the content of serum estradiol (E2), significantly downregulate the expression of ki-67 protein in mammary tissues, and inhibit mammary gland hyperplasia. CONCLUSION: XRSJ treatment can relieve mammary tissue hyperplastic lesions, reduce E2 levels and downregulate the expression of ki-67. It has a significant therapeutic effect on mammary gland hyperplasia.

[Intestinal bacteria metabolism of TSDP and characterization of metabolites in rats].

OBJECTIVE: To study the rat intestinal bacteria metabolism of total saponins of Dioscorea pathaica (TSDP) in vitro, and characterize the metabolites in serum and urine of rats after oral administration of TSDP 900 mg.kg-1. METHOD: TSDP metabolites were detected with thin-layer chromatography (TLC) and combination of electrospray ionization mass spectrometry (ESI-MS) and sequential tandem mass spectrometry (MSn). RESULT: In vitro, TSDP was decomposed easily by rat intestinal bacteria, and metabolites DP-1, DP-2, DP-4, DP-5 and diosgenin (Dio) were observed with prolongation of incubation time by ESI-MS2. In vivo, in the full-scan positive mass spectrum of the rat urine sample, the ion peak at m/z 415 (M-H) and its characteristic fragmentations at m/z 397 and m/z 271 in the MS/MS spectrum were identified with that of metabolite Dio, therefore metabolite Dio was deduced to exist in the rat urine, and metablite Dio was allso detected in the rat serum sample. CONCLUSION: TSDP is decomposed easily by rat intestinal bacteria and metabolite diosgenin is absorbed into blood after oral administration of TSDP.

Isoginsenoside-Rh3, a new triterpenoid saponin from the fruits of Panax ginseng C. A. Mey.

A new dammarane-type triterpene monoglucoside, named isoginsenoside-Rh(3), has been isolated from the fruits of Panax ginseng C. A. Mey, together with eight known analogs, ginsenoside-Rb(1), -Rb(2), -Rc, -Rd, -Re, -Rg(1), -Rh(1), -Rh(2). On the basis of chemical and physicochemical evidence, the structure of isoginsenoside-Rh(3) has been elucidated as 3-O-beta--glucopyranosyl-dammarane-(E)-20(22),24-diene-3beta,12beta-diol (1).