Small molecule proteomics quantifies differences between normal and fibrotic pulmonary extracellular matrices / 中华医学杂志(英文版)

BACKGROUND@#Pulmonary fibrosis is a respiratory disease caused by the proliferation of fibroblasts and accumulation of the extracellular matrix (ECM). It is known that the lung ECM is mainly composed of a three-dimensional fiber mesh filled with various high-molecular-weight proteins. However, the small-molecular-weight proteins in the lung ECM and their differences between normal and fibrotic lung ECM are largely unknown.@*METHODS@#Healthy adult male Sprague-Dawley rats (Rattus norvegicus) weighing about 150 to 200 g were randomly divided into three groups using random number table: A, B, and C and each group contained five rats. The rats in Group A were administered a single intragastric (i.g.) dose of 500 μL of saline as control, and those in Groups B and C were administered a single i.g. dose of paraquat (PQ) dissolved in 500 μL of saline (20 mg/kg). After 2 weeks, the lungs of rats in Group B were harvested for histological observation, preparation of de-cellularized lung scaffolds, and proteomic analysis for small-molecular-weight proteins, and similar procedures were performed on Group C and A after 4 weeks. The differentially expressed small-molecular-weight proteins (DESMPs) between different groups and the subcellular locations were analyzed.@*RESULTS@#Of the 1626 small-molecular-weight proteins identified, 1047 were quantifiable. There were 97 up-regulated and 45 down-regulated proteins in B vs. A, 274 up-regulated and 31 down-regulated proteins in C vs. A, and 237 up-regulated and 28 down-regulated proteins identified in C vs. B. Both the up-regulated and down-regulated proteins in the three comparisons were mainly distributed in single-organism processes and cellular processes within biological process, cell and organelle within cellular component, and binding within molecular function. Further, more up-regulated than down-regulated proteins were identified in most sub-cellular locations. The interactions of DESMPs identified in extracellular location in all comparisons showed that serum albumin (Alb) harbored the highest degree of node (25), followed by prolyl 4-hydroxylase beta polypeptide (12), integrin β1 (10), apolipoprotein A1 (9), and fibrinogen gamma chain (9).@*CONCLUSIONS@#Numerous PQ-induced DESMPs were identified in de-cellularized lungs of rats by high throughput proteomics analysis. The DESMPs between the control and treatment groups showed diversity in molecular functions, biological processes, and pathways. In addition, the interactions of extracellular DESMPs suggested that the extracellular proteins Alb, Itgb1, Apoa1, P4hb, and Fgg in ECM could be potentially used as biomarker candidates for pulmonary fibrosis. These results provided useful information and new insights regarding pulmonary fibrosis.

Blast-Induced Traumatic Brain Injury Triggered by Moderate Intensity Shock Wave Using a Modified Experimental Model of Injury in Mice / 中华医学杂志(英文版)

<p><b>Background</b>The increasing frequency of explosive injuries has increased interest in blast-induced traumatic brain injury (bTBI). Various shock tube models have been used to study bTBI. Mild-to-moderate explosions are often overlooked because of the slow onset or mildness of the symptoms. However, heavy gas cylinders and large volume chambers in the model may increase the complexity and danger. This study sought to design a modified model to explore the effect of moderate explosion on brain injury in mice.</p><p><b>Methods</b>Pathology scoring system (PSS) was used to distinguish the graded intensity by the modified model. A total of 160 mice were randomly divided into control, sham, and bTBI groups with different time points. The clinical features, imaging features, neurobehavior, and neuropathology were detected after moderate explosion. One-way analysis of variance followed by Fisher's least significant difference posttest or Dunnett's t 3-test was performed for data analyses.</p><p><b>Results</b>PSS of mild, moderate, and severe explosion was 13.4 ± 2.2, 32.6 ± 2.7 (t = 13.92, P < 0.001; vs. mild group), and 56.6 ± 2.8 (t = 31.37, P < 0.001; vs. mild group), respectively. After moderate explosion, mice showed varied symptoms of malaise, anorexia, incontinence, apnea, or seizure. After bTBI, brain edema reached the highest peak at day 3 (82.5% ± 2.1% vs. 73.8% ± 0.6%, t = 7.76, P < 0.001), while the most serious neurological outcomes occurred at day 1 (Y-maze: 8.25 ± 2.36 vs. 20.00 ± 4.55, t = -4.59, P = 0.048; 29.58% ± 2.84% vs. 49.09% ± 11.63%, t = -3.08, P = 0.008; neurologic severity score: 2.50 ± 0.58 vs. 0.00 ± 0.00, t = 8.65, P = 0.016). We also found that apoptotic neurons (52.76% ± 1.99% vs. 1.30% ± 0.11%, t = 57.20, P < 0.001) and gliosis (2.98 ± 0.24 vs. 1.00 ± 0.00, t = 14.42, P = 0.021) in the frontal were significantly higher at day 3 post-bTBI than sham bTBI.</p><p><b>Conclusions</b>We provide a reliable, reproducible bTBI model in mice that can produce a graded explosive waveform similar to the free-field shock wave in a controlled laboratory environment. Moderate explosion can trigger mild-to-moderate blast damage of the brain.</p>

Effect of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomi compatibility on uric acid metabolism and urinary neutrophil gelatinase-associated lipocalin and kidney injury molecule-1 in rats with hyperuricemia / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the effects of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomi compatibility (PR) on uric acid metabolism and the expression of urinary neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) in rats with hyperuricemia.</p><p><b>METHODS</b>Seventy male Sprague Dawley (SD) rats were randomly divided into 7 groups with 10 rats per group, including the normal group, model group, allopurinol group, benzbromarone group and PR groups at 3 doses (3.5, 7, 14 g/kg). Except the normal group, rats of the other groups were intragastrically administered 100 mg/kg hypoxanthine and 250 mg/kg ethambutol, and subcutaneously injected with 200 mg/kg potassium oxonate. All rats were continuously modeled for 17 days, and gavaged with corresponding drugs. The rats of the normal and model groups were gavaged with saline, once a day, for 2 weeks. The levels of serum uric acid (SUA), blood urea nitrogen (BUN) and creatinine (Cr) were determined. In addition, the contents of NGAL and KIM-1 in urine and the mRNA and protein expressions of xanthine oxidase (XOD) in liver of hyperuricemia rats were measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. Moreover, the pathological changes of kidney were analyzed by hematoxylin and eosin (HE) stain method.</p><p><b>RESULTS</b>Compared with the normal group, the levels of SUA, BUN, NGAL and KIM-1 and the expressions of hepatic XOD mRNA and protein in the hyperuricemia rats were increased signifificantly (P<0.01). PR signifificantly decreased the levels of SUA, BUN, NGAL and KIM-1 and down-regulated the mRNA and protein expressions of hepatic XOD (P<0.05 or P<0.01). In addition, the pathological changes of kidney were signifificantly suppressed by oral administration of PR.</p><p><b>CONCLUSIONS</b>PR ameliorated uric acid metabolism and protected renal function, the underlying mechanism was mediated by decreasing the levels of SUA, BUN, NGAL and KIM-1, inhibiting the expression of hepatic XOD and ameliorating the pathological change of kidney.</p>

Clinical characteristics of 638 patients infected with hepatitis C virus by post blood transfusion,in Shanghai / 中华流行病学杂志

Objective To investigate me clinical-epidemiologic characteristics of patients with hepatitis C virus(HCV)infection by post blood transfusion.Methods Polymerase chain reaction (PCR)and enzynle linked immunosorbent assay(ELISA)were used to detect HCV RNA and anti.HCV,respectively.Analysis was performed on patients'age distribution,cause of primary diseases,years ofexposure,ingredient and amount of transfusion,incubation period,disorder on liver function and changes on abdominal ultrasound image,etc.Results HCV RNA levels were higher than 3.0log10 copy/ml in 90.8%infected patients、with a median as 6.10 log10 copy/ml.19.2%of the patients showed viral load 3.0 to 4.0 iog10 copy/ml,and 66.1%of them showed 5.0 to 6.0 log10 copy/ml.Only 14.7%of the infected persons had HCV RNA levels higher than 7.0 log10 copy/ml.Eighty-one point five percent(44/54)of the infected persons were confirmed as HCV RNA positive by HCV RNA qualitative analysis with HCV genotype as primarily type 1.99.8%(636/637)of the pmients were detected as anti-HCV positive by serological test.The sensitivity of serological test was higher than both quantitative and qualitative HCV RNA assays(P=0.000,P=0.000,respectively).HCV infection post blood transfusion was more seen in common people at 40 to 60 years old Most cases(85.7%)had their first exposure during 1990 to 1994.More than 10% of the cases had primary diseases aS obstetrics,orthopedics or gastrointestinal tract hemorrhage.79.9%of the patients received whole blood product transfusion.The mean interval between transfusion and clinical diagnosis was 8.5±5.5 years.90.1%of the infected patients had liver function damage,while most of them showed elevated alanine aminotransferase(ALT)no more than 5 upper limits of normal(ULN).wheteas Serum total bilirubin(TBIL).ALT and aspartate aminotransferase(AST)≥5×ULN level were showing more clinicaI manifestations(P=0.000.P=0.001,P=0.009,respectively).Abdominal ultrasound among 8.9%of the infected persons showed changes in cirrhosis,and most of them werc older than 50years of age.Conclusion Most of the post transfusion HCV infected cases happened in adulthood,and were mainly exposed during 1990 to 1994.Infected pmients usually had their liver function damaged with elevated ALT no more than 5×ULN and with medium HCV RNA levels.HCV genotype was mainly for type 1.Patients who weTe ofolder age showed higher incidence ofcirrhosis.If a patients'infection period Was longer than 5 years,he/she would show higher incidence of cirrhosis.

Radiosensitivity of nasopharyngeal carcinoma in relation to cell cycle synchronization effect of tumor necrosis factor alpha / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of tumor necrosis factor alpha (TNFalpha) on radiosensitivity of nasopharyngeal carcinoma (NPC) in relation to TNFalpha-induced cell cycle synchronization.</p><p><b>METHODS</b>The radio-resistance of a NPC cell line subclone CNE-2Z-S1 was verified by in vivo experiments and flow cytometry was performed to evaluate cell cycle synchronization in TNFalpha-treated CNE-2Z-S1 cells. The radiosensitivity of the cell synchronized CNE-2Z-S1 cells was determined by clone formation in vitro and in vivo experiment in nude mice.</p><p><b>RESULTS</b>TNFalpha was capable of inducing cell cycle arrest and synchronization of CNE-2Z-S1 cells. Pretreatment with TNFalpha remarkably enhanced the radiosensitivity of CNE-2Z-S1 in vitro, and in vivo experiments with nude mice also suggested the role of TNFalpha in enhancing the radiosensitivity of NPC.</p><p><b>CONCLUSION</b>TNFalpha can enhance the radiosensitivity of NPC cells by inducing cell cycle synchronization.</p>

Study on the correlation between gene ponA/penA and the penicillin-resistance of Neisseria gonorrhoeae / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between penA/ponA and penicillin resistance of Neisseria gonorrhoeae.</p><p><b>METHODS</b>Agar dilution method was used to determine the minimum inhibitory concentrations (MICs) of the strains. Polymerase chain reaction-single stand conformation polymerphism (PCR-SSCP) and PCR- restriction fragment length polymorphism (RFLP) were used to detect the mutations in ponA and penA genes, which encoding the penicillin binding protein-1 and -2 (PBP1 and PBP2), respectively.</p><p><b>RESULTS</b>All the 80 N. gonorrhoeae isolates had a D345 insertion detected in penA while 93.7% of N. gonorrhoeae isolates having a point mutation Leu421 --> Pro in ponA. Most of the penicillinase producing N. gonorrhoeae (PPNG) strains possessed the mutations in ponA and penA.</p><p><b>CONCLUSION</b>Our data suggested that the plasmid and chromosome mediated penicillin-resistance conjugately increased the level of resistance.</p>

Modulation of frequency receptive field plasticity in rat auditory cortical neurons by electrical stimulation of medial prefrontal cortex / 生理学报

Using conventional electrophysiological technique, we investigated the effects of stimulating the medial prefrontal cortex (mPFC) on plasticity of frequency receptive field (RF) in auditory cortical (AC) neurons in rats. When the mPFC was electrically stimulated, the RF plasticity of 51 (27.2%) neurons was not affected and that of 137 neurons (72.8%) was either inhibited (71 neurons, 37.7%) or facilitated (66 neurons, 35.1%). The modulation of RF plasticity by the stimulation of mPFC was dependent upon the time interval between acoustic and electrical stimuli. The best interval time that produced optimal modulation (inhibition or facilitation) ranged from 5 to 30 ms. The inhibitory modulation of mPFC prolonged RF shifting time and shortened RF recovery time. Conversely, the facilitatory modulation of mPFC shortened RF shifting time and prolonged RF recovery time. Our results suggest that the mPFC may affect the plasticity of functional activity in AC neurons, and also may participate in the process of auditory learning and memory.

Correspondence analysis on random amplified polymorphic DNA genotyping and drug-resistance of Neisseria gonorrhoeae strains in Pudong area, Shanghai / 中华流行病学杂志

<p><b>OBJECTIVE</b>Using molecular epidemiology methods to investigate relationship between genotypes and drug-resistance of neisseria (N.) gonorrhoeae in Shanghai area.</p><p><b>METHODS</b>A random amplified polymorphic DNA (RAPD) fingerprint method at the molecular level was used to differentiate the strains which were isolated from the outpatients of sexually transmitted disease clinics. The sensitivity to antibiotic of the 78 N. gonorrhoeae strains on 9 different antibiotics was tested and the relationship between different genotypes and phenotypes was studied.</p><p><b>RESULTS</b>Selected RAPD primer could give out a group of amplification polymerase chain reaction bands with some main segments common to all the N. gonorrhoeae strains tested and some segments were different among the N. gonorrhoeae strains. All the 78 N. gonorrhoeae strains could be classified as three different groups (I, II and III). The strains could also be distinguished as four types (A, B, C and D) according to drug-resistance status. Using correspondence analysis method, the relationship between the three genotypes and four resistance types could be identified.</p><p><b>CONCLUSION</b>RAPD fingerprint seemed a useful genotyping method and could be used for molecular epidemiological studies.</p>

Application of random amplification polymorphic DNA in the genotyping of Neisseria gonorrhoeae / 中华流行病学杂志

<p><b>OBJECTIVE</b>To set up random amplified polymorphic DNAs (RAPD) method in genotyping Neisseria gonorrhoeae on DNA level, and to explore its use to trace the source of infection.</p><p><b>METHODS</b>Four different pretreatments were used to extract the Neisseria gonorrhoeae genomic DNA with its advantages and disadvantages compared. Arbitrary sequence was then used to amplify the genomic DNA of Neisseria gonorrhoeae and RAPD fingerprint maps was applied to distinct the Neisseria gonorrhoeae strains. Finally, RAPD fingerprint of Neisseria gonorrhoeae strain between patient and his/her sexual partner was compared.</p><p><b>RESULTS</b>Cetyltrimethylammonium bromide method was classical in extracting genomic DNA, and could get integrated genomic DNA and good fingerprint maps, since main segments were common to all the Neisseria gonorrhoeae but some were different among strains so that the fingerprint of different Neisseria gonorrhoeae were distinctive. However, fingerprint maps of Neisseria gonorrhoeae collected from sex partners were quite similar.</p><p><b>CONCLUSION</b>Based on genomic levels, effective fingerprint maps could be identified and to classify the Neisseria gonorrhoeae into different genotypes. RAPD fingerprint maps could be used to trace the source of infection.</p>

A case-control study on drinking green tea and decreasing risk of cancers in the alimentary canal among cigarette smokers and alcohol drinkers / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the role of green tea in decreasing the risks of gastric cancer, liver cancer, esophageal cancer among alcohol drinkers or cigarette smokers.</p><p><b>METHODS</b>A population based case-control study was conducted in Taixing, Jiangsu province.</p><p><b>RESULTS</b>In Taixing city, identified cases of stomach, liver and esophageal cancers were chosen with informed consent. The numbers were 206, 204, 218 respectively. Controls were chosen from normal population having lived in the area for longer than 10 years, also with informed consent. Green tea drinking seemed to have decreased 81%, 78%, 39% risk for the development of gastric cancer, liver cancer and esophageal cancer among alcohol drinkers. It might also have decreased 16%, 43%, 31% on the risks of developing the three kinds of cancers among cigarette smokers. Interaction assessment showed that drinking green tea could significantly decrease the risk of gastric cancer and liver cancer among alcohol drinkers, with ORs of interaction item 0.23 (95% CI: 0.10 - 0.55) and 0.25 (95% CI: 0.11 - 0.57) respectively.</p><p><b>CONCLUSION</b>Habit of drinking green tea seemed to have significant protective effects on the development of both gastric and liver cancer among alcohol drinkers while, green tea also having some protective effect on esophageal cancer among alcohol drinkers and on three kinds of cancers among cigarette smokers.</p>

Role of transferrin in the stimulation of Na,K-ATPase induced by low K+ in Madin Darby canine kidney cells / 生理学报

The presence of serum in a culture medium makes it impossible to identify whether changed cellular functions are directly caused by a manipulation itself or mediated by a component in serum. Madin Darby canine kidney cells can survive in a serum-free medium for about 48 h. We took this advantage to examine whether low K(+)-induced up-regulation of Na,K-ATPase requires serum. We found that serum was essential for low K(+) to induce an increase in Na,K-ATPase binding sites as quantified by ouabain factor binding assays. In an attempt to identify which component was critical, we screened EGF, IGF1, PGE1 and transferrin to identify which one can replace serum. We discovered that transferrin was the single most important factor that mimicked about 80% to 90% of the effect of serum. Transferrin potentiated the effect of low K(+) on the Na,K-ATPase binding sites in a time- and dose-dependent manner. Furthermore, transferrin was also required for low K(+)-induced increase in alpha(1)-promoter activity, alpha(1)- and beta(1)-subunit protein abundance of the Na,K-ATPase. In the presence of transferrin, low K(+) enhanced cellular uptake of iron approximately by 70%. Inhibition of intracellular iron activity by deferoxamine (30 micromol/L) abrogated the effect of low K(+). We conclude that stimulation of the Na,K-ATPase by low K(+) is critically dependent on transferrin. The effect of transferrin is mediated by increased iron transport.

Detection of COL1A1/PDGFB fusion transcripts in dermatofibroscoma protuberans by revers transcriptase-polymerase chain reaction using paraffin-embedded tissues / 中华病理学杂志

<p><b>OBJECTIVE</b>To detect the COL1A1/PDGFB fusion transcripts and discuss its clinicopathological significance in dermatofibroscoma protuberans.</p><p><b>METHODS</b>Formalin fixed, paraffin-embedded tumor specimens from 12 patients with DFSP were reviewed by light microscope and the expression of COL1A1/PDGFB mRNA resulting from the reciprocal translocation t(17;22) (q22;q13.1) was detected by one-step revers transcriptase-polymerase chain reaction. The following tumor specimens were included as controls: 2 fibrosarcoma, 2 malignant fibrous histocytoma, 3 leiomyosarcoma, 1 dermarofibroma and 1 nerve shealth tumor.</p><p><b>RESULTS</b>The COL1A1/PDGFB fusion transcripts were detected in 8 (67%) of 12 samples from patients with DFSP. Nucleotide sequence analysis using the PCR products confirmed that different regions of the COL1A1 gene, respectively, were fused with of PDGFB gene. No COL1A1/PDGFB fusion transcripts were detected in the control tumors.</p><p><b>CONCLUSION</b>Detection of specific COL1A1/PDGFB fusion transcripts in DFSP will help to diagnose the nature of DFSP and research the mechanism of its molecular histogenesis.</p>