Objective:To evaluate the effect of long-term intake of ω-3 polyunsaturated fatty acids (ω-3 PUFAs) on the activation of hippocampal microglia in a mouse model of postoperative cognitive dysfunction (POCD).Methods:Ninety-six clean-grade healthy male C57BL/6 mice, aged 8 weeks, weighing 18-24 g, were stratified according to body weight and divided into 4 groups ( n=24 each) by a random number table method: control diet group (group C), ω-3 PUFAs group (group ω), control diet plus POCD group (group C+ P) and ω-3 PUFAs plus POCD group (group ω+ P). Mice were fed a special ω-3 PUFAs diet (DHA 0.14 g/100 g, EPA 0.03 g/100 g) for 12 weeks in group ω and group ω+ P, while mice were fed with a control diet for 12 weeks in group C and group C+ P.Tibial fracture procedures were performed under isoflurane anesthesia to develop the POCD model after 12 weeks of feeding.The fear conditioning test and Y maze test were performed on 1st and 3rd days after developing the model.The mice were sacrificed after behavioral tests, and the hippocampal tissues were removed for determination of the contents of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) (by gas chromatography-mass spectroscopy), density of Iba-1 positive microglia (by immunofluorescence staining), and expression of mature brain-derived neurotrophic factor (mBDNF) and precursor brain-derived neurotrophic factor (pro-BDNF) (by Western blot), and contents of interleukin-1beta (IL-1β) and interleukin-6 (IL-6) (by enzyme-linked immunosorbent assay). Results:Compared with group C, the contents of DHA and EPA were significantly increased, the percentage of freezing time in the contextual test was increased, mBDNF/pro-BDNF ratio was increased ( P<0.05), no significant change was found in the rotation accuracy in Y maze test, density of Iba-1 positive microglia and contents of IL-1β and IL-6 in hippocampus ( P>0.05) in group ω ( P<0.05), and no significant change was found in the contents of DHA and EPA ( P>0.05), the percentage of freezing time in the contextual test and accuracy of rotation in Y maze test were decreased on 1st and 3rd days after operation, the density of Iba-1 positive microglia and contents of IL-1β and IL-6 were increased, and mBDNF/pro-BDNF ratio was decreased in group C+ P ( P<0.05). Compared with group C+ P, the contents of DHA and EPA were significantly increased, the percentage of freezing time in the contextual test and accuracy of rotation in Y maze test were increased on 1st and 3rd days after operation, the density of Iba-1 positive microglia and contents of IL-1β and IL-6 were decreased, and mBDNF/pro-BDNF ratio was increased in group ω+ P ( P<0.05). Conclusions:Long-term intake of ω-3 PUFAs can improve cognitive function in a mouse model of POCD, and the mechanism may be related to inhibition of activation of hippocampal microglia, reduction of inflammatory responses, and thus increasing the mBDNF/Pro-BDNF ratio.

【Objective】 To investigate the mechanism of pristane inducing autophagy in macrophages. 【Methods】 Pristane was used to stimulate NR8383， a rat macrophage cell line. The changes in signaling pathways of AMPK， mTOR， and endoplasmic reticulum （ER） stress pathways including eIF2α and IRE1α in the cell model， as well as the expression of transcriptional factor TFEB and its translocation to the nucleus， were detected by using Western blotting. ER stress pathways were intervened by using an inducer DTT or an inhibitor 4-PBA to determin its effect on mTOR expression and autophay. 【Results】 In pristane-stimulated NR8383 cell model， ER stress pathway eIF2α was activated at 0.5 h after stimulation， and then mTOR expression was decreased at 1 and 3 h after stimulation. There was no change for AMPK and IRE1α pathways. With 4-PBA treatment， pristane-reduced mTOR expression and increased LC3-II were reversed， while with DTT treatment， mTOR expression decreased and LC3-II expression increased even more. Pristane induced the expression and activation of TFEB in NR8383 cells. 【Conclusion】 Pristane induces ER stress and leads to autophagy enhancement in rat macrophages.

The glymphatic system is a fluid dynamics network that is important for maintaining homeostasis of the brain, and it is also a new target for the treatment of various central nervous system diseases. The crucial point regarding research into the glymphatic system is the microhydrodynamics of the cerebrospinal fluid tracer. This review summarizes the emerging technologies, such as magnetic resonance technology, two photon microscopic imaging technology, near infrared fluorescence imaging technology, and transcranial macroscopic imaging, and summarizes its research applications and technical advantages to provide methodological strategies for basic and clinical research on glymphatic system function.

【Objective】 To establish a microbial limit test method for diatomite and pearlite, and verify its applicability. 【Methods】 According to the requirements of general rule 1105, Microbial Limit Test for Non Sterile Products of Pharmacopoeia of the People′s Republic of China (2020 Edition), the applicability test of microbial counting methods for three batches of perlite and diatomite was conducted before the microbial limit test of samples. The microbial growth of filter aid was analyzed and the recovery rate of each test bacterium was calculated. 【Results】 The ratio of the colony number of the test group minus the colony number of the test sample control group to the bacterial liquid control group was in the range of 0.5~2.0. 【Conclusion】 The method is accurate, reliable and can be used for microbial limit test of diatomite and perlite.

OBJECTIVE：To study ongoing change characteristics of the contents of syringin and total flavonoids in different medicinal parts （root bark ，tree bark ，leaf）of Toricellia angulata from Guizhou ，and to provide reference for the development and application of T. angulata . METHODS ：The root bark ，tree bark and leaf parts of T. angulata during different harvesting periods （Jan.-Dec.） were taken as the research samples. The content of syringin was determined by HPLC. The determination was performed on Agela Promosil C 18 column with mobile phase consisted of 0.5％ phosphoric acid solution-acetonitrile （gradient elution）at the flow rate of 1.0 mL/min. The detection wavelength was set as 210 nm，and column temperature was 35 ℃. The sample size was 5 μL. The content of total flavonoids was determined by UV-visible spectrophotometry under detection wavelength of 510 nm. RESULTS ：The linear range of syringin and total flavonoids were 0.095 9-1.150 8 mg/mL（r＝0.999 6）and 0.072 2- 1.083 0 mg/mL（r＝0.999 9），respectively. RSDs of precision ，stability and repeatability tests were all less than 3％（n＝6）. The average recoveries were 101.74％（RSD＝2.36％ ，n＝6）and 99.63％（RSD＝2.19％ ，n＝6），respectively. During different harvesting periods ，the contents of syringin in root bark ，tree bark ，leaf of T. angulata collected on Aug. ，May and Sept. were the highest，and the contents of total flavonoids in samples collected on Feb. ，Dec. and Sept. were the highest. The contents of syringin in different medicinal parts of T. angulata were in descending order as follows as tree bark ＞root bark ＞leaf，and the content of syringin was commonly relatively high in tree bark part ；the content of total flavonoids in different medicinal parts of T. angulata were in descending order as follows as root bark ＞tree bark ＞leaf，and the contents of total flavonoids in three medicinal parts was generally low. The content of total flavonoids in root bark was the highest in Feb. of that year ，and the content of syringin in root bark at same month was second only to Aug. of that year ；the content of syringin in tree bark was the highest in May ，and the content of total flavonoids in tree bark at same month was second only to Oct. and Dec. of that year ；the contents of total flavonoids and syringin in leaf were the highest in Sept. of that year. CONCLUSIONS ：It is suggested that Feb. is the best time for harvesting root bark ，May for tree bark and Sept. for leaf of T. angulata .

Leukocyte differentiation antigens (LDAs) play important roles in the immune system, by serving as surface markers and participating in multiple biological activities, such as recognizing pathogens, mediating membrane signals, interacting with other cells or systems, and regulating cell differentiation and activation. Data mining is a powerful tool used to identify novel LDAs from whole genome. LRRC25 (leucine rich repeat-containing 25) was predicted to have a role in the function of myeloid cells by a large-scale "omics" data analysis. Further experimental validation showed that LRRC25 is highly expressed in primary myeloid cells, such as granulocytes and monocytes, and lowly/intermediately expressed in B cells, but not in T cells and almost all NK cells. It was down-regulated in multiple acute myeloid leukemia (AML) cell lines and bone marrow cells of AML patients and up-regulated after all-trans retinoic acid (ATRA)-mediated granulocytic differentiation in AML cell lines and acute promyelocytic leukemia (APL; AML-M3, FAB classification) cells. Localization analysis showed that LRRC25 is a type I transmembrane molecule. Although ectopic LRRC25 did not promote spontaneous differentiation of NB4 cells, knockdown of LRRC25 by siRNA or shRNA and knockout of LRRC25 by the CRISPR-Cas9 system attenuated ATRA-induced terminal granulocytic differentiation, and restoration of LRRC25 in knockout cells could rescue ATRA-induced granulocytic differentiation. Therefore, LRRC25, a potential leukocyte differentiation antigen, is a key regulator of ATRA-induced granulocytic differentiation.
Antigens, Differentiation ; immunology ; metabolism ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Granulocytes ; cytology ; drug effects ; immunology ; metabolism ; Humans ; Leukocytes ; cytology ; drug effects ; immunology ; metabolism ; Membrane Proteins ; antagonists & inhibitors ; immunology ; metabolism ; RNA, Small Interfering ; pharmacology ; Tretinoin ; pharmacology

Background and purpose:The previous research has found that the prostate stromal cells derived from different prostate zones have distinct effect on prostate epithelial cells. We also revealed that LMO2 protein was highly expressed in PZ stromal cells (PZSCs) and prostate cancer associated fibroblasts (CAFs) compared with TZ stromal cells. This study investigated the effect of LMO2 protein in prostate stromal cells on proliferation and invasion of prostate cancer PC-3 cells and its mechanisms. Methods:Lentivirus overexpression vectors were used to establish LMO2-overexpressed prostate WPMY-1 stromal cell line. shRNA plasmids were used to suppress LMO2 in CAFs. LMO2 mRNA and protein level of both WPMY-1 and CAFs were evaluated by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Then, PC-3 cells were co-cultured with different prostate stromal cells and the in vitro proliferation and invasion of PC-3 were measured by CCK-8 and matrigel invasion assays respectively. Results:When co-cultured with LMO2-overexpressed prostate stromal cells, both proliferation and in-vasion of PC-3 were improved. However, when co-cultured with CAFs which have inhibited expression of LMO2, the proliferation and invasion of PC-3 were reduced. The protein array proifling found that both interleukin-11 (IL-11) and ifbroblast growth factor-9 (FGF-9) were enhanced extensively in the supernatant collected from LMO2-overexpressed WPMY-1 cells. Conclusion:The expression of LMO2 in prostate stromal cells could be responsible for development of prostate cancer. Paracrine of cytokines, such as IL-11 and FGF-9, from LMO2-overexpressed stromal cells had effects on the proliferation and invasion of prostate cancer cells.

Objective To evaluate the characteristics of the testicular torsion patients with the first symptom of acute abdominal pain.Methods From October 1998 to May 2014, 8 adolescent or boys with testicular torsion presenting acute abdominal pain instead of scrotal pain as their primary symptom were retrospectively reviewed.The average age was 12 (3-16) years.An acute abdominal pain presented firstly in all cases, which accompanied by nausea and vomiting in 4 cases, groin pain in 2 cases, fever in 1 case. In genital examination, the scrotum skin was red and swollen, while the testis was in transverse position and tenderness.The testicular and homolateral abdominal pain got worse when we pulled the involved testis. Doppler ultrasound examination revealed that the blood supply of the involved testis decreased or disappeared.Results In the 8 cases, 5 cases were not examined their testicles or scrotum during the first evaluation, which were found the testicular necrosis during the following exploratory surgery, and orchidectomy was performed. In the other 3 cases, external genital were examined during the first evaluation, and the scrotal and testicular abnormalities were detected.Doppler ultrasonography showed abnormality in testicular blood supply.Testicular torsion was proved during the emergent operation, and the 3 testes were salvaged.The mean duration time from symptom onset to operation was 4 h in the salvaged group, while the average time was 28 h in the orchiectomy group.The mean follow-up time was 18 ( 10-36) months.In the 3 cases whose testis was saved, 1 affected testis was atrophied, and the blood supply of the other two was good.No severe complications occurred in the other 5 cases.Conclusions Testicular torsion should be considered in adolescents and boys with acute lower abdominal pain.The routine external genital examination should be performed when a boy or adolescent complains of abdominal pain.

Objective To detect the changes of G protein-coupled inwardly rectifying potassium channels (GIRK)expression in allergic asthma model and identify the regulatory factors.Methods The E3 rat asthma models were induced by challenge with ovalbumin 14 days after immunization with ovalbumin and aluminium adjuvant.The asthma models were evaluated based on changes in lung pathomorphology and total IgE levels.The levels of GIRK1-4 mRNA and protein were detected using real time-PCR and Western blot.The anatomic sites where GIRK was expressed dominantly in the lung were identified using immunohistological staining.To identify the effects of IL-4 on the expressions of GIRK channels,GIRK 1 -4 mRNA and protein in IL-4 stimulated bronchial epithelial cell line A549 were detected by RT-PCR and Western blot.Results The levels of GIRK1-4 mRNA and protein decreased significantly in the lung in asthmatic E3 rats.The results of immunohistological staining showed that GIRK channels were dominantly expressed in airway epithelia in the lung.The levels of GIRK 1-4 mRNA and protein were down-regulated in time-and dose-dependent manners in IL-4 treated A549.Conclusion IL-4 down-regulates the expression levels of GIRK subunits in bronchial epithelia during allergic asthma.

OBJECTIVETo screen molecular markers in early breast cancer and establish gene subtyping-based diagnostic criteria for predicting the prognosis of early breast cancers.

METHODSTumor tissue specimens were obtained from 8 patients with early breast cancer for analysis of the differentially expressed genes using Agilent custom 8×15 000 chips in combination with the prognostic data of the patients. Another 42 tumor tissue specimens were used to validate the differential genes by real-time fluorescent quantitative PCR.

RESULTSGene microarray analysis identified 132 differentially expressed genes between the patients with favorable and poor prognosis, and 44 of these genes were significantly up-regulated (by over two folds) and 88 down-regulated in patients with poor prognoses.

CONCLUSIONThe gene expression profiles differ in early breast cancer tissues of the same pathological type but with different clinical stages and prognoses, and CD44, MKI67, NTRK2, Nek2, C16orf60, TOP2A, ANCCA, and RRM2 genes can be used as the prognostic markers for early breast cancer.

Adult ; Aged ; Biomarkers, Tumor ; analysis ; genetics ; Breast Neoplasms ; genetics ; pathology ; Carcinoma, Ductal, Breast ; genetics ; pathology ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Middle Aged ; Neoplasm Staging ; Oligonucleotide Array Sequence Analysis ; Prognosis