[Gene expression signature analysis of peripheral blood mononuclear cells from patients with for high altitude pulmonary hypertension and value for potential drug selection].

Objective: To investigate the gene expression characteristics of peripheral blood mononuclear cells from patients with high altitude pulmonary hypertension (HAPH) in Naxi residents living in Lijiang, Yunnan, and to explore the underlying pathogenesis and value for potential drug selection. Methods: This is a case-control study. Six patients with HPAH (HPAH group) and 4 normal subjects (control group) were selected from the Naxi residents who originally lived in Lijiang, Yunnan Province. The general clinical data of the two groups were collected, and the related indexes of pulmonary artery pressure were collected. Peripheral blood mononuclear cells of the subjects were collected for RNA sequencing. The differences on gene expression, regulatory network of transcription factors and drug similarity between the two groups were compared. The results were compared with the public data of idiopathic pulmonary arterial hypertension (IPAH). Biological processes and signal pathways were analyzed and compared between HPAH and IPAH patients. Results: The age of 6 patients with HAPH was (68.1±8.3) years old, and there were 2 males (2/6). The age of 4 subjects in the control group was (62.3±10.9) years old, and there were 2 males (2/4). Tricuspid regurgitation velocity, tricuspid pressure gradient and pulmonary systolic pressure in HAPH group were significantly higher than those in control group (all P<0.05). The results of RNA sequencing showed that compared with the control group, 174 genes were significantly upregulated and 169 genes were downregulated in peripheral blood mononuclear cells of HAPH group. These differentially expressed genes were associated with 220 biological processes, 52 molecular functions and 23 cell components. A total of 21 biological processes and 2 signal pathways differed between HPAH and IPAH groups, most of which were related to inflammation and immune response. ZNF384, SP1 and STAT3 were selected as highly correlated transcription factors by transcription factor prediction analysis. Trichostatin A and vorinostat were screened out as potential drugs for the treatment of HAPH by drug similarity analysis. Conclusions: There are significant differences in gene expression in peripheral blood monocytes between HAPH patients and normal population, and inflammation and immune dysfunction are the main pathogenic factors. Trichostatin A and Vorinostat are potential drugs for the treatment of HAPH.

[Clinical study of psychological changes and post traumatic stress disorder in elderly patients with hip fracture].

Objective: To explore the psychological state and affected factors of elderly patients with hip fractures. Methods: A retrospective analysis of 156 elderly hip fracture patients(>65 years) admitted to the Department of Orthopaedics, the First Affiliated Hospital of Harbin Medical University from January 2016 to August 2019 was performed. General and psychological information were collected by questionnaire.General information included age, gender, education, whether surgery, length of stay.SCL-90, a self-assessment scale, was chosen as the psychological test to analyzed the elderly hip fracture patients' psychological status during hospitalization and the norms of SCL-90 in Chinese which were established in 1986 were used as the control group. The prognostic factors were examined by univariate and multivariate analysis. Results: Somatization, interpersonal sensitivity, depression, anxiety, paranoid factor scores, and total scores of the elderly hip fracture patients were significantly higher than control group(all P=0.00).Univariate analysis and logistic regression analysis showed that non-surgery treatment and more than 10 days of hospitalization were independent prognostic factors that affected the psychological state of elderly hip fracture patients (all P=0.00). Conclusion: Elderly patients hospitalized with osteoporosis and hip fractures are prone to have negative emotional and psychological changes.The length of hospitalization and the choice of treatment can affect patients' psychological state, suggesting that effective psychological intervention is necessary.

Meningoencephalitis or meningitis in relapsing polychondritis: four case reports and a literature review.

Relapsing polychondritis (RP) is a rare autoimmune disease that affects cartilage throughout the body, causing episodic and progressive inflammation. Although rare, RP has diverse acute and subacute nervous system complications, which may sometimes precede systemic manifestations. Here, we report four patients with RP who presented with meningoencephalitis or meningitis without infectious aetiology. In addition, we review the literature for this disease with regard to clinical manifestations and treatment options.

Role of circadian clocks in the development and therapeutics of cancer.

Circadian clocks are endogenous molecular time-keeping systems that underlie daily fluctuations in multiple physiological and biochemical processes. It is now well recognized that dysfunction of the circadian system may be associated with a heightened incidence of cancer. This brief review presents evidence supporting the important role played by circadian clocks in the development of cancer and the therapeutic efficacy of anticancer agents. A number of circadian clock genes have been identified, which include transcription factors that regulate gene expression. Continued research in this area should increase understanding of the role of circadian clocks which could, ultimately, reduce the incidence of cancer in people with disrupted sleep-wake cycles, such as 24-h shift workers and flight attendants, and provide optimal chronopharmacology for cancer treatment.

Computed tomography in left-sided and right-sided blunt diaphragmatic rupture: experience with 43 patients.

AIM: To investigate differences in the radiographic signs for left and right-sided blunt diaphragmatic rupture (BDR) in order to provide guidance to avoid missing these injuries. MATERIALS AND METHODS: A retrospective review of the computed tomography (CT) examinations of 43 patients with BDR treated at our hospital between January 1995 and 2007 was undertaken. The presence of diaphragmatic discontinuity, diaphragmatic thickening, herniation of abdominal organs into the thoracic cavity, collar/hump sign, dependent viscera sign, abnormally elevated 4 cm or more above the dome of the other-sided hemi-diaphragm, and of associated injuries was recorded and their relationship to each other and to BDR diagnosis examined. A comparison between the use of axial and sagittal/coronal reconstruction images in diagnosis was also performed in 15 patients. RESULTS: On axial imaging, left-sided diaphragmatic rupture occurred in 31 patients (72%) and right-sided in 12 (28%). Twenty-nine patients had associated injuries. More than 60% of the patients showed the "dependent viscera" sign, "abdominal organ herniation" sign, diaphragm thickening, or had a more than 4 cm elevation of one side of the diaphragm. "Diaphragmatic discontinuity" and "stomach herniation" were seen almost exclusively in left-sided rupture. Those with BDR and haemothorax had a significantly lower incidence of "diaphragm discontinuity" (p=0.034) than those without haemothorax. Sagittal/coronal reconstruction slightly increased the number of band signs, diaphragmatic discontinuities and diaphragmatic thickenings seen. CONCLUSIONS: Of the CT signs examined in this study, when herniation of abdominal organs was used as a diagnostic marker, only a very small fraction of trauma patients identifiable by CT would be missed. Further, CT signs differ for left-sided and right-sided BDR, thus the possibility of BDR should be considered when any of the reported CT signs are present.

Giant extracranial aneurysms of both internal carotid arteries with aberrant jugular veins in a patient with neurofibromatosis type 1.

Although neurofibromatosis type 1 (NF-1) is commonly considered neurocutaneous, severe arterial and venous abnormalities have been noted. Our patient, a 28-year-old woman, had bilateral giant extracranial aneurysms of the internal carotid arteries as well as skull base meningoceles involving the jugular foramina and aberrant jugular veins. CT and MR imaging, as well as digital subtraction and/or other angiography techniques, may be required to clarify pathology in patients with suggested vascular lesions.

Vertebral osteonecrosis: MR imaging findings and related changes on adjacent levels.

BACKGROUND AND PURPOSE: No prior report has comprehensively discussed the intravertebral vacuum cleft sign and the fluid sign on MR images of vertebral osteonecrosis. The purpose of this study was to investigate MR images of osteonecrotic vertebral bodies and adjacent intervertebral disks and vertebral bodies. METHODS: We retrospectively reviewed MR images of patients with vertebral osteonecrosis. Affected vertebral bodies with osteonecrosis were defined as an avascular area (nonenhanced area on enhanced T1-weighted images) with collections of intravertebral fluid (hyperintense signal on T2-weighted images), air (signal void on all images), or both. The degree of vertebral collapse was classified as mild (>50%) or severe (<50%) preserved vertebral height. Changes in adjacent intervertebral disks or vertebral bodies 2 above and 2 below the affected vertebrae were compared. RESULTS: We enrolled 112 patients (30 men, 82 women; 121 vertebral bodies) in our study. Intravertebral air alone was observed in 48 involved levels (39.7%), intravertebral fluid alone was found in 47 (38.8%), and both coexisted in 26 (21.5%). Degree of vertebral collapse in affected vertebral bodies significantly differed with presence of air or fluid (P < .05). Vertebral compression fractures adjacent to the affected vertebral bodies were more common in those with intravertebral air alone than in those with intravertebral fluid alone (P < .05). CONCLUSION: Vertebral collapse was more advanced and adjacent vertebral compression fractures were more frequent in patients with intravertebral air than in those with intravertebral fluid.

Comparison of agar plate and real-time PCR on enumeration of Lactobacillus, Clostridium perfringens and total anaerobic bacteria in dog faeces.

AIMS: To compare agar plate and real-time PCR methods on enumeration of total anaerobic bacteria, Lactobacillus and Clostridium perfringens in dog faeces. METHODS AND RESULTS: Thirty-two faecal specimens from Labrador retriever dogs were used to compare agar plate and real-time PCR enumeration methods for Lactobacillus, C. perfringens and total anaerobic bacteria. Total anaerobic bacteria, C. perfringens and Lactobacillus of faeces were counted (as CFU g(-1) faeces) for 48-h incubation at 37 degrees C in an anaerobic gas chamber on genus-selective media. Total genomic DNA from samples was extracted by the QIAamp DNA stool mini kit. The quantification of DNA (as DNA copy per gram faeces) by real-time PCR was performed with a LightCycler system with the QuantiTect SYBR green PCR kit for PCR amplification. The results indicated that there was a significant correlation between CFU and DNA copy of Lactobacillus (R2 = 0.78, P < 0.01) and total anaerobic bacteria (R2 = 0.21, P < 0.05); but no correlation was found between CFU and DNA copy of C. perfringens. The regression equations for Lactobacillus and total anaerobic bacteria were log(DNA copy) = 0.83 x log(CFU) + 1.43 and log(DNA copy) = 1.62 x log(CFU) - 6.32 respectively. CONCLUSIONS: The real-time PCR method could be used to enumerate Lactobacillus within 2 days when compared with plating method which requires 5-6 days. SIGNIFICANCE AND IMPACT OF THE STUDY: The real-time PCR method and the primer set for Lactobacillus spp. harboured in the dog intestine can be used for rapid enumeration of lactobacilli and monitoring of the faecal Lactobacillus community.

Planar heterogeneous structures for coherent emission of radiation.

The coupling of excited surface polaritons with thermal radiation through diffraction by gratings results in coherent thermal emission in a certain frequency range toward well-defined directions for p polarization. A planar coherent source is proposed that uses multilayers of negative permittivity (epsilon) and negative permeability (mu) materials. Owing to the excitation of surface polaritons at the interface between the negative-epsilon and negative-mu layers, coherent emission can be achieved for both s and p polarization. Moreover, one can control the emission frequency and direction by adjusting the layer thicknesses.

Fructooligosaccharide supplementation in the yearling horse: effects on fecal pH, microbial content, and volatile fatty acid concentrations.

Short-chain fructooligosaccharides (FOS) were supplemented to the diets of nine quarter horses ranging in age from 489 to 539 d with initial BW averaging 400.6 +/- 21.2 kg. The objectives of this study were to determine the effects of dietary FOS on the fecal responses in terms of pH, the microbial population, and VFA concentrations. The horses were used in a 3 x 3 replicated Latin square design, fed according to NRC requirements, and their individual diets were supplemented with no FOS (CON), 8 g of FOS/d (LOW), or 24 g of FOS/d (HIGH) over three 10-d feeding periods. On the last 3 d of each 10-d feeding period, a single fecal sample was collected between 0730 and 0930. Fecal pH decreased linearly (P = 0.01) from 6.48 with the CON diet to 6.38 with the HIGH diet, but there was no change (P = 0.19 for linear effect) in fecal consistency among treatments. A quadratic effect (P < 0.01) was observed for fecal Escherichia coli population, but no difference (P = 0.88 for linear effect) was found in fecal Lactobacilli enumeration among treatments. The presence of fecal Bifidobacteria was unable to be confirmed and was therefore not reported. Fecal acetate concentrations increased linearly (P = 0.03), with means of 2.13, 2.18, and 2.52 mg/g of wet feces for CON, LOW, and HIGH treatments, respectively. Similarly, fecal propionate concentrations increased linearly (P = 0.01), with means of 0.58, 0.64, and 0.73 mg/g for CON, LOW, and HIGH treatments, respectively. Fecal butyrate concentrations also increased linearly (P = 0.02), with means of 0.40, 0.46, and 0.54 mg/g for CON, LOW, and HIGH treatments, respectively. Total VFA (P = 0.01) and lactate (P = 0.02) concentrations increased linearly, with total VFA means of 3.47, 3.69, and 4.25 mg/g for CON, LOW, and HIGH treatments, respectively, and lactate means of 0.36, 0.41, and 0.47 mg/g for CON, LOW, and HIGH treatments, respectively. Supplementing FOS in diets fed to yearling horses altered fecal microbial populations, fecal VFA concentrations, and pH.

Pre-harvest factors influencing the acid resistance of Escherichia coli and E. coli O157:H7.

The effects of pH, acetate, propionate, or butyrate concentration, and diet on acid resistance of fecal Escherichia coli and E. coli O157:H7 were determined by in vitro and in vivo experiments. The pH tested was from 4.0 to 8.0, and the VFA concentrations tested were 0 to 100 mM. The E. coli O157:H7 used was strain 505B. In an in vivo study, cattle were fed a grain-based diet, then either not switched or switched to a grain-based diet with 3% added calcium carbonate or two fiber-based diets (soybean hulls or hay). Acid resistance was expressed as viability after acid-shock at pH 2.0 for 1 h and 4 h for fecal E. coli and E. coli O157:H7, respectively. Enumeration methods used were multitube fermentation, agar plate, and petri-film methods. The E. coli O157:H7 was not found in continuous culture inocula or in vivo samples. The viability of fecal E. coli decreased linearly (P < 0.01) as the culture pH increased, and viability of E. coli O157:H7 was highest (P < 0.01) when cultivated at pH 6.0. The viability of fecal E. coli and E. coli O157:H7 showed quadratic responses (P < 0.05) as acetate and butyrate concentrations increased at pH 7.2, with maximal acid resistance at 20 and 12 mM, respectively. As propionate concentration increased, the acid resistance was not different (P > 0.05) for fecal E. coli. Acid resistance of E. coli was induced by acetate and butyrate, even though the environmental pH was near neutral. Similar results were measured in the in vivo study, where viability after acid shock was more dependent on VFA concentration than on pH. Increasing the dietary calcium carbonate concentration also increased (P < 0.05) acid resistance of fecal E. coli. Results from these studies demonstrated that culture pH and VFA affect acid resistance of E. coli.

[Changes of human recombination bone morphogenetic protein-2 in bone and marrow in tail suspended rats].

OBJECTIVE: To study human recombination bone morphogenetic protein-2 (rhBMP-2) change in bone and marrow under simulated weightlessness. METHOD: 5 SD rats were tail suspended for 14, 28 d with another 5 freely active rats as control. Histological samples were in situ hybridized. RESULT: rhBMP-2 expression of bone and marrow were lower in tail suspended rats than control levels (P < 0.05). rhBMP-2 expression tail suspended rats in 14 day were higher than that in 28 day suspended group (P < 0.05). CONCLUSION: rhBMP-2 levels in rats bone and marrow were lower after tail suspension.

Ruminal peptide concentration required to optimize microbial growth and efficiency.

Experimentation, with a single-phase continuous culture system operated at fractional dilution rates of 0.03 and 0.09 per hour and four cannulated crossbred steers (260 +/- 20 kg) used in a 4 x 4 Latin square design, was conducted to determine the level of ruminally degradable protein (RDP) that maximized microbial efficiency (MOEFF), microbial nitrogen flow, and nutrient digestibility in the rumen. Treatments consisted of increasing diet RDP levels (from 3.5 to 17.5% RDP on a DM basis). The basal diet was corn with casein or soybean meal used as the source of RDP in the continuous culture experiment and animal study, respectively. Dietary nonstructural carbohydrate (NSC) ranged from 52 to 75% on a dry matter basis. Urea was added to ensure an adequate ammonia nitrogen source for microbial growth. The RDP levels did not affect true digestibility of dry matter and organic matter, ruminal pH, particulate passage rate, or liquid passage rate in the animal study. As RDP increased, bacterial nitrogen production increased linearly (P < 0.05) only for the 0.09/h dilution rate. Microbial efficiency was not influenced by RDP level. Ruminal peptide and NH3 N concentration increased linearly (P < 0.05) as RDP increased. Based on this experimentation, 1.8 mM peptide maximized MOEFF when ammonia nitrogen was not limiting (> 2 mg/dL). We concluded from this research that the RDP requirement of NSC-fermenting bacteria was lower than that currently fed in many diets.

In vitro metabolism and interaction of cilostazol with human hepatic cytochrome P450 isoforms.

1. Cilostazol (OPC-13013) undergoes extensive hepatic metabolism. The hydroxylation of the quinone moiety of cilostazol to OPC-13326 was the predominant route in all the liver preparations studies. The hydroxylation of the hexane moiety to OPC-13217 was the second most predominant route in vitro. 2. Ketoconazole (1 microM) was the most potent inhibitor of both quinone and hexane hydroxylation. Both the CYP2D6 inhibitor quinidine (0.1 microM) and the CYP2C19 inhibitor omeprazole (10 microM) failed to consistently inhibit metabolism of cilostazol via either of these two predominant routes. 3. Data obtained from a bank of pre-characterized human liver microsomes demonstrated a stronger correlation (r2=0.68, P < 0.01) between metabolism of cilostazol to OPC-13326 and metabolism of felodipine, a CYP3A probe, that with probes for any other isoform. Cimetidine demonstrated concentration-dependent competitive inhibition of the metabolism of cilostazol by both routes. 4. Kinetic data demonstrated a Km value of 101 microM for cilostazol, suggesting a relatively low affinity of cilostazol for CYP3A. While recombinant CYP1A2, CYP2D6 and CYP2C19 were also able to catalyze formation of specific cilostazol metabolites, they did not appear to contribute significantly to cilostazol metabolism in whole human liver microsomes.

Simultaneous quantitative determination of cilostazol and its metabolites in human plasma by high-performance liquid chromatography.

A high-performance liquid chromatographic (HPLC) method for the simultaneous determination of cilostazol, a quinolinone derivative, and its known metabolites OPC-13015, OPC-13213, OPC-13217, OPC-13366, OPC-13269, OPC-13326 and OPC-13388 in human plasma was developed and validated. Cilostazol, its metabolites and two internal standards, OPC-3930 and OPC-13112, were extracted from human plasma by a combination of liquid-liquid and liquid-solid phase extractions, with combined organic solvents of n-butanol, methanol, chloroform, methyl-tert.-butyl ether, and a Sep-Pak silica column. The combined extract was then evaporated and the residue was reconstituted in ammonium acetate buffer (pH 6.5). The reconstituted solution was injected onto a HPLC system and was subjected to reversed-phase HPLC on a 5 microm ODS-80TM column to obtain quality chromatograph and good peak resolution. A gradient mobile phase with different percentages of acetonitrile in acetate buffer (pH 6.5) was used for the resolution of analytes. Cilostazol, its metabolites and the two internal standards were well separated at baseline from each other with resolution factor being 74 and 138. This HPLC method was demonstrated to be specific for all analytes of interest with no significant interference from the endogenous substances of human plasma. The lower limit of quantitation was 20 ng/ml for cilostazol and all metabolites. The method was validated initially for an extended linear range of 20-600 ng/ml for all metabolites and cilostazol, and has been revised later for a linear range of 20-1200 ng/ml for cilostazol and two major and active metabolites OPC-13015 and OPC-13213. The overall accuracy (relative recovery) of this method was established to be 98.5% to 104.9% for analytes with overall precision (CV) being 1.5% to 9.0%. The long-term stability of clinical plasma samples was established for at least one year at -20 degrees C. Two internal standards of OPC-3930 and OPC-13112 were evaluated and validated. However, the data indicated that there was no significant difference for all accuracy and precision obtained by using either OPC-3930 or OPC-13112. OPC-3930 was chosen as the internal standard for the analysis of plasma samples from clinical studies due to its shorter retention time. During the validation standard curves had correlation coefficients greater than or equal to 0.998 for cilostazol and the seven metabolites. These data clearly demonstrate the reliability and reproducibility of the method.

[Effect of ligustrazine on isolated myocardial ischemic reperfusion injury in rats].

In recent years, it is believed by some scholars that the injury of myocardial ischemic reperfusion is correlated to the thromboxane A2 (TXA2) released by platelets. In order to explore that whether the myocardial and hemangio-endothelial cells participate in the TXA2 production during the process of reperfusion, the modified Langendorff method was used to establish the model of reperfusing the isolated rat heart. On the other hand, this experiment was also intended to observe the effect of ligustrazine on the injury of myocardial ischemic reperfusion. The results revealed that the level of thromboxane B2 (metabolite of TXA2) and lactic dehydrogenase (LDH) in coronary sinus reflux fluid increased during the process of reperfusion, while the level of 6-keto-PGF1 alpha in the same fluid relatively decreased (P < 0.05). The ratio of TXB2/6-keto-PGF1 alpha was raised. The ligustrazine inhibited the release of TXB2 and LDH, but promoted the production of 6-keto-PGF1 alpha (P < 0.05). The results also proved that the myocardial and hemangio-endothelial cells could synthesize TXA2, and the amount of TXA2 released increased during the reperfusion of ischemic myocardium, which was likely to be the major factor of the injury of ischemic myocardial reperfusion. Ligustrazine plays an important role in protecting the myocardium.

The pharmacokinetics of aspirin in rats and the effect of buffer.

Aspirin (acetylsalicyclic acid) was administered to rats intravenously, orally, and intraintestinally at different doses or in different dosage forms. The distribution and elimination kinetics of aspirin in rats following intravenous administration were best described by a two-compartmental open system and were dose independent up to 15 mg/kg. The terminal elimination half-life following intravenous dosing (10 mg/kg) was 3.36 +/- 0.85 min (n = 15) with the clearance being 8.40 +/- 1.24 L/(kg.hr). Intravenous distribution and elimination kinetics of aspirin in rats were not influenced by an orally administered buffered solution with a buffer capacity of 0.933 mEq ANC (acid neutralizing capacity) per kg of body weight. However, this orally buffered solution did change the gastrointestinal absorption kinetics of aspirin in rats. The absolute bioavailable dose of aspirin was 56.6 +/- 10.4% (n = 6) following its administration in an unbuffered solution while it was only 31.8 +/- 8.0% (n = 6) following administration in the buffered solution. The corresponding values of the absolute bioavailable doses were 43.4 +/- 3.7% and 25.5 +/- 1.8% following intraintestinal administration. The lower systemic availability of aspirin in the presence of buffer is attributed to a greater fraction of the administered dose becoming available for absorption from the intestine where the extraction efficiency is higher than that in the stomach.