BACKGROUNDAdrenocorticotrophin (ACTH)-secreting pituitary adenomas account for approximately 7% - 14% of all pituitary adenomas, but its pathogenesis is still enigmatic. This study aimed to explore mechanisms underlying the pathogenesis of ACTH-secreting pituitary adenomas.

METHODSWe used fiber-optic beadarray to examine gene expression in three ACTH-secreting adenomas compared with three normal pituitaries. Four differentially expressed genes from the three ACTH-secreting adenomas and three normal pituitaries were chosen randomly for validation by reverse transcriptase-real time quantitative polymerase chain reaction (RT-qPCR). We then analyzed the differentially expressed gene profile with Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway.

RESULTSFiber-optic beadarray analysis showed that the expression of 28 genes and 8 expressed sequence tags (ESTs) were significantly increased and the expression of 412 genes and 31 ESTs were significantly decreased. Bioinformatic and pathway analysis showed that the genes HIGD1B, EPS8, HPGD, DAPK2, and IGFBP3 and the transforming growth factor (TGF)-β signaling pathway and extracellular matrix (ECM)-receptor interaction pathway may play important roles in tumorigenesis and progression of ACTH-secreting pituitary adenomas.

CONCLUSIONSOur data suggest that numerous aberrantly expressed genes and several pathways are involved in the pathogenesis of ACTH-secreting pituitary adenomas. Fiber-optic beadarray combined with pathway analysis of differential gene expression appears to be a valid method of investigating tumour pathogenesis.

ACTH-Secreting Pituitary Adenoma ; etiology ; genetics ; Adenoma ; etiology ; genetics ; Adult ; Disease Progression ; Expressed Sequence Tags ; Extracellular Matrix Proteins ; physiology ; Female ; Fiber Optic Technology ; Gene Expression Profiling ; Humans ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; methods ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; physiology ; Transforming Growth Factor alpha ; physiology

AIMTo study the effects of TNF receptor blocking peptide on adjuvant arthritis in rats.

METHODSThe model of rat adjuvant arthritis was induced by injection of complete Freund's adjuvant. The TNF receptor blocking peptide was injected locally in the ankle. The ankle swelling, the pathologic changes in the ankle joint and the expression of IL-1 beta mRNA and TNF-alpha mRNA by peritoneal macrophages (RT-PCR) were observed.

RESULTSThe model of rat adjuvant arthritis induced by injection of complete Freund's adjuvant was similar to human rheumatoid arthritis. The treatment with TNF receptor blocking peptide for 10 days resulted in complete inhibition of joint swelling, a decrease in infiltration of inflammatory cell into joint tissue, an obvious alleviation of inflammatory pathological damages and an apparent decline of TNF-alpha mRNA and IL-1 beta mRNA of peritoneal macrophages of rats.

CONCLUSIONThe TNF receptor blocking peptide can protect the joint from inflammatory damage induced by adjuvant arthritis by suppression of TNF-alpha and IL-1 production, thereby alleviating the pathological injury of joint and controlling effectively the clinic course of arthritis.

Animals ; Ankle Joint ; pathology ; Arthritis, Experimental ; immunology ; pathology ; Interleukin-1 ; biosynthesis ; genetics ; Macrophages, Peritoneal ; metabolism ; Male ; Peptides ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Tumor Necrosis Factor ; antagonists & inhibitors ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVETo establish a stable method for obtaining large quantity of highly purified immature dendritic cells (imDCs) in vitro, and identify the morphology, function and surface markers of the cells.

METHODSCD117(+) hemopoietic stem cells (HSCs) were isolated and purified from the bone marrow of healthy C57 mice by magnetic affinity cell sorting. After cell expansion by treatment with stem cell factor (SCF) and interleukin-3 (IL-3), the HSCs were induced for directional differentiation into imDCs by treatment with GM-CSF, IL-4 and IL-10. The imDCs obtained were identified by morphological and functional observation under inverted microscope, scanning electron microscope and transmission electron microscope, followed by detection of the expressions of the surface markers using flow cytometry.

RESULTSAfter 3, 5 and 7 days of culture in the presence of SCF+IL-3, the cells were expanded by 10.34-/+1.43, 22.65-/+2.71 and 54.39-/+3.08 folds, respectively. The HSCs were successfully induced to differentiate into imDCs with phagocytotic activity. The dendrites of the imDCs were short small, and appearing spinous. The expressions of surface markers were detected from the cells showing the phenotype of CD11c(+), I-A/I-E(low), CD40(-), CD80(-), CD86(-).

CONCLUSIONThe method described allows steadily acquisition of large quanty of highly purified imDCs and of their effective identification in vitro.

Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Dendritic Cells ; cytology ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Proto-Oncogene Proteins c-kit

BACKGROUNDRestoration of blood flow to the ischemic liver lobes may paradoxically exacerbate tissue injury, which is called hepatic ischemia/reperfusion injury (IRI). Toll-like receptor 4 (TLR4), expressed on several liver cell types, and the nuclear factor-kappa B (NF-kappaB) signaling pathway are crucial to mediating hepatic inflammatory response. Because IRI is essentially a kind of profound acute inflammatory reaction evoked by many kinds of danger signals, we investigated TLR4/NF-kappaB signaling pathway activation in a murine model of partial hepatic IRI.

METHODSWild-type mice (WT, C3H/HeN) or TLR4 mutant mice (C3H/HeJ) were subjected to 45 minutes of partial hepatic ischemia followed by 1 hour, 3 hours of reperfusion. Sham group accepted the same procedure without the obstruction of blood supply. At the end of reperfusion, the compromise of liver function and the histological change of liver sections were measured as the severity of liver injury. The level of endotoxin in the portal vein was measured by limulus assay. NF-kappaB activation was determined by electrophoretic mobility shift assay (EMSA). The levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in systemic blood after hepatic IRI were assessed by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe compromise of liver function and the morphological injuries in mutant mice were relieved more markedly than those in WT mice after partial hepatic IRI. NF-kappaB activation in WT mice was stronger than that in TLR4 mutant mice, and both were stronger than those in the sham operated mice (P < 0.01). Endotoxin in each group was undetectable. The levels of TNF-alpha and IL-1beta in systemic blood were elevated in both strains, but lower in the sham operated group. These mediators were significantly decreased in TLR4 mutant mice compared with those in WT mice (P < 0.01).

CONCLUSIONSThe TLR4/NF-kappaB signaling pathway may mediate hepatic IRI triggered by endogenous danger signals. Inhibition of the TLR4/NF-kappaB pathway may be a potential therapeutic target for attenuating ischemia/reperfusion-induced tissue damage in some clinical settings.

Alanine Transaminase ; blood ; Animals ; Interleukin-1beta ; biosynthesis ; Liver ; blood supply ; Mice ; Mice, Inbred C3H ; NF-kappa B ; physiology ; Reperfusion Injury ; etiology ; Signal Transduction ; physiology ; Toll-Like Receptor 4 ; physiology ; Tumor Necrosis Factor-alpha ; biosynthesis

BACKGROUNDGlobally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered 'nurse cells' in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro.

METHODSAdult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets' function when they were co-cultured for 21 days in vitro.

RESULTSIn cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P > 0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P < 0.01).

CONCLUSIONSA method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets' function when they were co-cultured in vitro. They could be a helper cell in islet transplantation.

Adult ; Cell Separation ; methods ; Cell Survival ; Cells, Cultured ; Coculture Techniques ; Humans ; Islets of Langerhans ; physiology ; Islets of Langerhans Transplantation ; Male ; Sertoli Cells ; cytology ; physiology

Objective To explore and compare the relevant regional anatomies as they relate the fron- tolateral keyhole approach under microscopy and neuroendoscopy for operations in anterior cranial base and sellar region.Methods Fifteen silieone-injected cadaveric heads were dissected to reveal and compare the extent of expesure through the transfrontolateral keyhole approach under neuroendoscopy and microscopy. Results Portions in the areas of olfactory groove,sellar region and sylvian tissure were blind under micro- scope.Endoscope could allow observation of areas considered blind under the microscope.It could increase light intensity during the approach to objects,extend viewing angles,clear depiction of details in close-up po- sitions and inspect hidden structures.But images of endoscope were two dimensional,lack of view depth.Mi- croscopy and neuroendoscopy could help each other to recuperate deficiency.Conclusion Endoscope-assis- ted neuromicrosurgery is helpful,safe and minimally invasive to treat deepseated lesions in anterior cranial base,sellar region by transfrontolateral keyhole approach.

OBJECTIVETo evaluate the chromium (Cr) levels in blood and urine among general population in China between 2009 and 2010, and thereby to analyze its prevalent features.

METHODSFrom year 2009 to 2010, a total of 11 983 subjects of general population aged between 6 and 60 year-old were recruited from 24 districts in 8 provinces in eastern, central and western China mainland, by cluster random sampling method. The information about their living environment and health status were collected by questionnaire, and 11 983 blood samples and 11 853 urine samples were also collected. Inductively coupled plasma mass spectrometry (ICP-MS) was applied to test the Cr level both in blood and urine; and the Cr distribution in blood and urine among groups of population in different ages, genders and districts, were then analyzed.

RESULTSAmong general population in China, the geometric mean (GM) of Cr concentration in blood was 1.19 µg/L, with median at 1.74 µg /L and 95% percentile at 5.59 µg/L. The Cr concentration in blood among males and females were separately 1.18 µg/L and 1.20 µg/L(P > 0.05); while its GM in the groups of population aged 6 - 12, 12 - 16, 16 - 20, 20 - 30, 30 - 45 and 45 - 60 years old were 1.00, 1.22, 1.01, 1.40, 1.27 and 1.30 µg/L (P < 0.01), respectively; and the figures in populations from eastern, central and western China were 1.00, 1.70 and 1.98 µg/L (P < 0.01), respectively. Among general population, the GM of Cr concentration in urine was 0.53 µg/L, with median was lower than 0.42 µg/L and 95% percentile at 3.53 µg/L. The Cr concentration in urine among males and females were separately 0.52 µg/L and 0.53 µg/L (P > 0.05);while its GM in the groups of population aged 6 - 12, 12 - 16, 16 - 20, 20 - 30, 30 - 45 and 45 - 60 years old were 0.56, 0.60, 0.52, 0.50, 0.52 and 0.46 µg/L (P < 0.01), respectively;and the figures in populations from eastern, central and western China were 0.58, < 0.42 and 0.60 µg/L (P < 0.01), respectively.

CONCLUSIONThe study reported the Cr levels in blood and urine among general population in China, and thereby provided basic data evidence for the following Cr biological monitoring studies in near future.

Adolescent ; Adult ; Child ; China ; Chromium ; blood ; urine ; Female ; Humans ; Male ; Middle Aged ; Population Surveillance ; Young Adult

ObjectiveTo evaluate the analgesic effect of intrarectal local anesthesia (IRLA) versus that of periprostatic nerve block anesthesia (PPNB) in initial transrectal ultrasound-guided prostate biopsy (TRUS-PB) for patients with different prostate volumes (PV).

METHODSA total of 253 patients undergoing initial TRUS-PB in our hospital from January 2014 to November 2017 were divided into three PV groups (<50 ml, 50－100 ml, and >100 ml), each again randomized into three subgroups (control, IRLA, and PPNB) with the random number table method. The pain during the procedure was assessed based on the Visual Analogue Scale (VAS) scores and the blind method was used by the biopsy operator, VAS valuator and data analyst.

RESULTSAmong the patients with PV <50 ml, the VAS scores in the blank control, IRLA, and PPNB subgroups were 4.39±0.87, 3.51±0.84 and 3.43±1.07, respectively, remarkably higher in the control than in the IRLA and PPNB groups (P<0.05), but with no statistically significant differences between the latter two (P>0.05). Among those with PV of 50－100 ml, the VAS scores in the three subgroups were 4.50±1.05, 4.38±1.13 and 3.38±1.44, respectively, markedly higher in the control and IRLA than in the PPNB group (P<0.05), but with no statistically significant differences between the former two groups (P>0.05). Among those with PV >100 ml, the VAS scores in the three subgroups were 5.19±1.05, 5.00±1.25 and 4.19±0.91, respectively, remarkably higher in the former two groups than in the latter (P<0.05), but with no statistically significant differences between the former two groups (P>0.05).

CONCLUSIONSEither IRLA or PPNB can be recommended for initial TRUS-PB in patients with PV <50 ml, PPNB for those with PV of 50－100 ml, and PPNB with other painkillers for those with PV >100 ml.

Administration, Rectal ; Aged ; Anesthesia, Local ; methods ; Anesthetics, Local ; administration & dosage ; Biopsy ; Humans ; Male ; Nerve Block ; methods ; Pain Measurement ; Pain, Procedural ; etiology ; prevention & control ; Prospective Studies ; Prostate ; pathology

Objective: To investigate the clinical application and outcomes of microdissection testicular sperm extraction (micro-TESE) in patients with nonmosaic Klinefelter syndrome (KS). METHODS: A total of 143 nonmosaic KS patients underwent micro-TESE in the Center of Reproductive Medicine of Peking University Third Hospital between July 2012 and August 2016. We analyzed their clinical and follow-up data and evaluated the outcomes. RESULTS: Spermatozoa were successfully retrieved from the testicular tissue in 44.76% (64/143) of the patients, 84.4% (54/64) by unilateral and 15.6% (10/64) by bilateral micro-TESE. Seventy-five of the KS patients were followed up in the years of 2014 and 2015. Of the 34 patients with successful sperm retrieval, 73.52% (25/34) achieved clinical pregnancy and 8 boys and 8 girls were already born in 14 of the 25 cases. CONCLUSIONS The micro-TESE is a useful method for sperm retrieval in nonmosaic KS patients, with high rates of sperm retrieval, clinical pregnancy, and birth of biological offspring.
Female ; Humans ; Klinefelter Syndrome ; Male ; Microdissection ; Pregnancy ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; Sperm Retrieval ; Spermatozoa ; Testis

A new styrene dimer derivative has been isolated from the branch of Litsea greenmaniana by column chromatography over silica gel and Sephadex LH-20, as well as semi-preparative HPLC. Its structure was identified by spectroscopic data analysis (MS, UV, IR, 1D and 2D NMR) as (E)-2,4-bis(p-hydroxyphenyl)-2-butenol, named as listeanol. At a concentration of 1×10-5 mol•L⁻¹, compound 1 was inactive in the assays, including cytotoxicity against human tumor cell lines (HCT-8, Bel-7402, BGC-823, A549 and A2780), antioxidant activity in Fe²⁺-cystine-induced rat liver microsomal lipid peroxidation, neuroprotective activity against serum deprivation or glutamate induced neurotoxicity in cultures of PC12 cells, and the inhibitory activity against protein tyrosine phosphatase 1B (PTP1B).