Investigation of Calcitonin and Calcitonin Gene-related Peptide in 88 Cases of Medullary Thyroid Carcinoma

Objective： To investigate the changes of calcitonin （CT） and calcitonin gene-related peptide （CGRP） in patients with medullary thyroid carcinoma （MTC）. Methods： Fifty-eight cases of MTC were selected and the relationship between the CT levels and metastasis was investigated. The immunohistochemical method was used to detect the expression of CT and CGRP in the 58 samples of MTC tissues. The CT and CGRP in 30 newly diagnosed MTC inpatients were measured before operation and in the first few days after operation using a radioimmunoassy. Results： （1） The rate of residual tumor had a significant difference between the normal serum CT group one month after operation and the elevated group at the same period （P〈0.01）. （2） Immunohistochemical study revealed the positive rate of CT was about 98%, and that of the CGRP was 87.8%. （3） Part of the patients had an elevated CGRP levels while CT levels was normal. （4） The serum CT levels were decreased to a stable range one week after operation. Conclusion： CT is a useful index to evaluate the efficacy of surgical treatment. The measurement of serum CGRP is helpful in the diagnosis of MTC, especially for those whose preoperative CT levels are normal.

Material Growth and Device Fabrication of GaN-Based Blue-Violet Laser Diodes

Studies on first GaN-based blue-violet laser diodes(LDs) in China mainland are reported.High quality GaN materials as well as GaN-based quantum wells laser structures are grown by metal-organic chemical vapor deposition method.The X-ray double-crystal diffraction rocking curve measurements show the full-width half maximum of 180″ and 185″ for (0002) symmetric reflection and (10 12) skew reflection,respectively.A room temperature mobility of 850cm2/(V·s) is obtained for a 3μm thick GaN film.Gain guided and ridge geometry waveguide laser diodes are fabricated with cleaved facet mirrors at room temperature under pulse current injection.The lasing wavelength is 405 9nm.A threshold current density of 5kA/cm2 and an output light power over 100mW are obtained for ridge geometry waveguide laser diodes.

A cadmium metallothionein gene of ridgetail white prawn Exopalaemon carinicauda(Holthuis,1950) and its expression

Metallothioneins （MTs） are a group of low molecular weight cysteine-rich proteins capable of binding heavy metal ions. A cadmium metallothionein （EcMT-Cd） cDNA with a 189 bp open reading frame （ORF） that encoded a 62 amino acid protein was obtained from Exopalaemon carinicauda. Seventeen cysteines were in the deduced amino acid sequence, and the cysteine （Cys）-rich characteristic was revealed in different metallothioneins in other species. In addition, the deduced amino acid sequence did not contain any aromatic amino acid residues, such as tyrosine （Tyr）, tryptophan （Trp）, and phenylalanine （Phe）. EcMT- Cd mRNA was expressed in all tested tissues （the ovary, muscle, stomach, and hepatopancreas）, and its expression profiles in the hepatopancreas were very different when shrimps were exposed to seawater containing either 50 pmol/L CuSO4 or 2.5 gmol/L CdC12. The expression of EcMT-Cd was significantly up-regulated in shrimp exposed to CuSO4 for 12 h and down-regulated in shrimps exposed to CdC12 for 12 h. After 24 h exposure to both metals, its expression was down-regulated. By contrast, at 48 h the EcMT-Cd was up-regulated in test shrimps exposed to CdC12. The transcript of EcMT-Cd was very low or even absent before the zoea stage, and the expression of EcMT-Cd was detected from mysis larvae-I, then its expression began to rise. In conclusion, a cadmium MT exists in E. carinicauda that is expressed in different tissues and during different developmental stages, and responds to the challenge with heavy metal ions, which provides a clue to understanding the function of cadmium MT

EFFECTS OF ACUPUNCTURE ON AMINO ACID CONTENTS IN CEREBROSPINAL FLUID AFTER BRAIN INJURY

Objective To probe into the mechanism on acupuncture treatment for brain injury. Methods Thirty cases of acute craniocerebral injury were divided into two groups according to the sequence of visiting. In the control （15 cases）, the routine western medicine was applied. In the experimental group （15 cases）, on the basis of routine western medicine, acupuncture was applied on Neiguan （ 内关 PC 6） and Zusanli （足三时 ST 36）. Before the treatment and on the 1^st and 5^th days after the treatment, the contents of aspartic acid （Asp）, glutamic acid （Glu） and 7-amino butyric acid （GABA） were observed successively and the relevant analysis was done. Results There was no significant difference in the concentrations of Asp, Glu and GABA before and on the 1^st day after the treatment between two groups （P〉0.05）. The difference had not been presented between the concentrations on the 1^st day and before the treatment. But, the difference was significant or very significant between the concentrations on the 5^th days and before the treatment （P〈0.05, P 〈 0.01 ）. In the experimental group, the concentration of Asp in the cerebrospinal fluid was lower obviously compared with that in the control （P〈0.05） and the concentration of GABA was higher compared with the control （P 〈 0.01）. Conclusion Acupuncture lowered the contents of Asp and Glu and increased the level of GABA in the cerebrospinal fluid rapidly so that the excitation and inhibition in the nervous system could be rebalanced. It was further indicated that acupuncture worked on the treatment of craniocerebral injury.

Transcript profiles of mitochondrial and cytoplasmic manganese superoxide dismutases in Exopalaemon carinicauda under ammonia stress

Superoxide dismutase （SOD） is one of the most important antioxidant defense enzymes, and is considered as the first line against oxidative stress. In this study, we cloned a mitochondrial manganese （Mn） SOD （mMnSOD） cDNA from the ridgetail white prawn Exopalaemon carinicauda by using rapid amplification of cDNA ends （RACE） methods. The fulMength cDNA for mMnSOD was 1 014-bp long, containing a 5＇-untranslated region （UTR） of 37-bp, a 3＇-UTR of 321-bp with a poly （A） tail, and included a 657-bp open reading frame encoding a protein of 218 amino acids with a 16-amino-acid signal peptide. The protein had a calculated molecular weight of 23.87 kDa and a theoretical isoelectric point of 6.75. The mMnSOD sequence included two putative N-glycosylation sites （NHT and NLS）, the MnSOD signature sequence 18~DVWEHAYY^87, and four putative Mn binding sites （H48, H96, D180, and H184）. Sequence comparison showed that the mMnSOD deduced amino acid sequence of E. carinicauda shared 97%, 95%, 89%, 84%, 82%, 72%, and 69% identity with that ofMacrobrachium rosenbergii, Macrobrachium nipponense, Fenneropeneaus chinensis, Callinectes sapidus, Perisesarma bidens, Danio rerio, and Homo sapiens, resectively. Quantitative real-time RT-PCR analysis showed that mMnSOD transcripts were present in all E. carinicauda tissues examined, with the highest levels in the hepatopancreas. During an ammonia stress treatment, the transcript levels of mMnSOD and cMnSOD were up-regulated at 12 h in hemocytes and at 24 h in the hepatopancreas. As the duration of the ammonia stress treatment extended to 72 h, the transcript levels of mMnSOD and cMnSOD significantly decreased both in hemocytes and hepatopancreas. These findings indicate that the SOD system is induced to respond to acute ammonia stress, and may be involved in environmental stress responses in E. carinicauda.

Iodothyronine deiodinase gene analysis of the Pacific oyster Crassostrea gigas reveals possible conservation of thyroid hormone feedback regulation mechanism in mollusks

Iodothyronine deiodinase catalyzes the initiation and termination of thyroid hormones(THs) effects, and plays a central role in the regulation of thyroid hormone level in vertebrates. In non-chordate invertebrates, only one deiodinase has been identified in the scallop C hlamys farreri. Here, two deiodinases were cloned in the Pacific oyster C rassostrea gigas( Cg Dx and C g Dy). The characteristic in-frame TGA codons and selenocysteine insertion sequence elements in the oyster deiodinase c DNAs supported the activity of them. Furthermore, seven orthologs of deiodinases were found by a tblastn search in the mollusk Lottia gigantea and the annelid C apitella teleta. A phylogenetic analysis revealed that the deiodinase gene originated from an common ancestor and a clade-specific gene duplication occurred independently during the differentiation of the mollusk, annelid, and vertebrate lineages. The distinct spatiotemporal expression patterns implied functional divergence of the two deiodinases. The expression of C g Dx and Cg Dy was influenced by L-thyroxine T4, and putative thyroid hormone responsive elements were found in their promoters, which suggested that the oyster deiodinases were feedback regulated by TH. Epinephrine stimulated the expression level of C g Dx and Cg Dy, suggesting an interaction effect between different hormones. This study provides the first evidence for the existence of a conserved TH feedback regulation mechanism in mollusks, providing insights into TH evolution.

Effects of propofol on Bax and Bcl-2 expression induced by tumour necrosis factor-α in mouse spinal cord neurons in vitro

Objective:To study the effects of propofol on Bax and Bcl-2 expression induced by tumour necrosis factor-α(TNF-α) in mouse spinal cord neurons in vitro. Methods:Spinal cord neurons were isolated from fetal mice and cultured in Neurobasal medium with B27 supplement. On the 7^th day, cultured neurons were randomly divided into 6 groups: control group, propofol (50 umol/L)group, TNF-α group, propofol (25 umol/L) with TNF-α group, propofol (50 umol/ L) with TNF-α group, and propofol ( 100 umol/L)with TNF-α group. Propofol was added to the cultured cells respectively and the cells were incubated for 30 min. Then TNF-α was added to the cultured cells(the final concentration of TNF-α was 2 000 U/ml) and incubated for 24 h. The Bax and Bcl-2 expression were measured by immunocytochemical technique. Results:In TNF-α group, the expression of Eax increased and the expression of Bcl-2 decreased (P<0.01, vs control). However, treatment with various concentrations of propofol (25, 50, 100 umol/L) decreased the expression of Bax and increased the expression of Bcl-2 (P<0.05, P<0.05, P<0.05, vs TNF-α). Conclusion: Propofol can inhibit the apoptosis induced by TNF-α by modulating the expression of Bax and Bcl-2.

Relationship belween Gene Expression of bFGF and Prolifercrtive Activity in Meningioma

OBJECTIVE To study the relationship between bFGF gene expression and proliferative activity in meningiomas. METHODS Thirty-seven samples of meningioma were examed using Northern hybridization for bFGF-gene expression and immunohistochemi- stry for bFGF protein expression and the Ki-67LI. RESULTS bFGF mRNA was detected in 22 meningiomas of Grade Ⅰ and in all samples of Grade Ⅱ and Ⅲ. Six of the Grade Ⅰ tumors were negative, giving an overall positive rate of 83.8 % for bFGF mRNA. Autoradiography was conducted using a thin scaning apparatus, bFGF mRNA expression compared to that for β-actin was 0.34±0.06 for Grade Ⅰ tumors compared to 0.82±0.12 for Grade Ⅱ and Ⅲ tumors. The difference was highly significant （P〈0.001）. The results of immunohistochemistry showed that the number of the positive cells for the bFGF protein was 29.7±7.6% in Grade Ⅰ tumors and 63.2±11.7 % in Grade Ⅱ plus Ⅲ, resulting in a significant difference between them （P〈0.001） . The Ki-67LI was found to be 2.8±1.1% in Grade Ⅰ and 6.5±1.3 % in Grade Ⅱ plus Ⅲ. The former was significantly lower than the later （P〈0.001）. CONCLUSION The expression of bFGF was correlated well with the malignancy of the meningiomas.

Construction of metastatic spinal cancer tissue microarrays

Objective： To explore the construction of metastatic spinal cancer （MSC） tissue microarrays and validate its value in immunohistochemical study of MSC. Methods： Paraffin-embedded specimens from 71 MSC cases and 6 primary tumor cases were selected as donor blocks and prepared into MSC tissue microarrays by tissue array arrangement, the steps of which included location, punching, sampling, sample seeding, and re-diagnosis by hematoxylin-eosin （HE） as well as MMP-9 and MMP-14 immunohistoehemical staining. Results： The MSC tissue microarrays thus constructed were intact and craekless, containing 154 complete and well arranged microarray points. None of the sectioned tissue microarrays was lost, and the results of HE staining was consistent with the primary pathologic diagnoses. Immunohistochemical staining was also good without non-specific or marginal effect. Conclusion： The MSC tissue microarrays have a high value in the immunohistochemical study of MSC.

Novel Functions of Interleukin-1 Beta in Spotted Halibut(Verasper variegatus) at Its Embryonic and Metamorphic Stages

Interleukin-lbeta （IL-1β） is a cytokine involved in the control of inflammatory responses. The functions of IL-1β have been extensively studied in many vertebrates. However, few studies focused on the functions of IL-1β in the spotted halibut （Verasper variegatus）, an endangered aquatic fish that easily suffers from bacterial and viral infections. We first cloned and sequenced the IL-1β gene from V. variegatus （VvIL-1β）. Its conservative interleukin-1 propeptide and interleukin-1/18 domains corresponding regions, characteristics of the IL-1 protein family and TA-rich motifs relating to inflammatory responses were similar to those of the IL-1β genes from other teleost and mammals. The phylogenetic analysis showed that VvIL-1β was homologous to the IL-1β gene of other vertebrates. Treatment with pathogen-associated molecular patterns （PAMPs） induced a significant and rapid up-regulation of VvIL-1β expression in the peripheral blood leukocytes （PBLs）, revealing that VvIL-1β is actively involved in the host ilmnune responses against bacterial and viral pathogens. The expression level of VvIL-1β was relatively high in the PBLs, gill, and intestine, indicating the immunological function of VvIL-1β . VvIL-1β also had interesting expression patterns at specific developmental stages, implying that the IL-1β of flatfishes may have some novel functions during embryonic development and metamorphosis. The 3D structure prediction also provided information about how VvIL-113 interacts with its receptor protein. These results suggested that VvIL-1β may have multiple functions in V. variegatus and provided us valuable information for understanding the novel functions of IL- 1β.

Changes in Global DNA Methylation Intensity and DNMT1 Transcription During the Aging Process of Scallop Chlamys farreri

DNA methylation is an important epigenetic regulatory mechanism that influences genomic stability, gene activation, X-chromosome inactivation and other factors. A change in DNA methylation is usually associated with aging and cellular senescence. DNA methyltransferase 1(DNMT1) is the most abundant DNA methyltransferase, and it plays an important role in maintaining the established methylation pattern during DNA replication in vertebrates. Although the effect of aging on DNA methylation has been well studied in vertebrates, little research has been conducted in invertebrates, especially in marine bivalves. In this study, we examined global DNA methylation levels in four groups of adult Zhikong scallop Chlamys farreri at different ages. The results showed that both the age and tissue type had a strong effect on the DNA methylation. In addition, a significant decrease in DNA methylation with aging(1–4 years) can be detected in mantle, kidney and hepatopancreas. We further measured the change in DNMT1 transcript abundance using quantitative reverse transcription PCR(q RT-PCR), which revealed that DNMT1 transcription significantly decreased with aging in mantle and hepatopancreas and strongly correlated with DNA methylation(R = 0.72). Our data provided greater insight into the aging-related decline of DNA methylation, which could aid in gaining a better understanding of the relationship between DNA methylation and the aging process in bivalve mollusks.

Division and correlation of Yixian Formation in western Liaoning,China

The Yixian Formation is a series of volcanic-sedimentary rocks in Biepiao area of Liaoning Province. It is mainly composed of basic and intermediate-basic volcanic lava,pyroclasts and terrestrial sedimentary rocks. Based on the regularity of volcanic activity,the Yixian Formation was divided by the present authors into four members in ascending order:the first member is of basal conglomerate,basic and intermediate-basic volcanic rocks; the second member is of lake phrase sedimentary rocks,or in another word,precious fossil-rich sedimentary beds; the third is of basic volcanic rocks; and the fourth is of upper conglomerate. Field mapping and comprehensive study also indicate that there are abundant vertebrate fossils (mainly of Psittacosaurus) in the first member of the Yixian Formation,and the Jehol Biota (including Sinosauropterxy,Confuciusorns sanctus,Archaefructus,etc.) is yielded in the second member of Yixian Formation. From west to east,the volcanic activity of Yixian Formation changed regularly from early to late,and from basic and intermediate-basic to acid (alkali).

Effect of temperature on the standard metabolic rates of juvenile and adult Exopalaemon carinicauda

Ridgetail white prawn （Exopalaemon carinicauda） are of significant economic importance in China where they are widely cultured. However, there is little information on the basic biology of this species. We evaluated the effect of temperature （16, 19, 22, 25, 28, 31, and 34℃） on the standard metabolic rates （SMRs） of juvenile and adult E. carinicauda in the laboratory under static conditions. The oxygen consumption rate （OCR）, ammonia-N excretion rate （AER）, and atornic ratio of oxygen consumed to nitrogen consumed （O：N ratio） of juvenile and adult E. carinicauda were significantly influenced by temperature （P〈0.05）. Both the OCR and AER of juveniles increased significantly with increasing temperature from 16 to 34℃, but the maximum OCR for adults was at 31℃. Juvenile shrimp exhibited a higher OCR than the adults from 19 to 34℃. There was no significant difference between the AERs of the two life-stages from 16 to 31 ℃ （P〉0.05）. The O：N ratio in juveniles was significantly higher than that in the adults over the entire temperature range （P〈0.05）. The temperature coefficient （Q_10） of OCR and AER ranged from 5.03 to 0.86 and 6,30 to 0.85 for the adults, respectively, and from 6,09-1.03 and 3.66-1.80 for the juveniles, respectively. The optimal temperature range for growth of the juvenile and adult shrimp was from 28 to 31℃, based on Q_10 and SMR values. Results from the present study may be used to guide pond culture production ofE. carinicauda.

Correlation of Cerebrospinal Fluid Sialic Acid with Various Enzymes in Patients with Meningitis

Meningitis is often associated with cerebral compromise which may be responsible for neurological squeal in nearly half of the survivors. Little is known about the mechanisms of Central Nervous System （CNS） involvement in bacterial meningitis. The present study was undertaken to assess the changes in biochemical parameters including glucose, protein, Sialic Acid （SA） and various enzymes like Aspartate Aminotransferase （AST）, Alanine Aminotransferase （ALT）, Creatine Kinase （CK）, Gamma Glutamyltranspeptidase 7 （GGTP） and Lactate Dehydrogenase （LDH） have been determined in the Cerebrospinal Fluid （CSF） of the meningitis patients （n = 26） and compared with control subjects （n = 26）. Total Protein, CK, SA （P 〈 0.02）, AST （P 〈 0.001） and ALT （P 〈 0.0001） of meningitis patients were significantly higher, whereas, glucose （P 〈 0.05） and Gamma Glutamyltranspeptidase （GGT） （P 〈 0.01 and P 〈 0.05, respectively） were significantly lower than those of the controls. This study suggests that loss of integrity of brain-CSF barriers, sialic acid and enzyme profiles may contribute to the severity and neurological complications of meningitis.

Recording of spared motor evoked potentials and its augmentation by 4-aminopyridine in chronic spinal cord-injured rats

Objective To research the direct electrophysiological evidence of discomplete spinal cord injury (SCI) and the effect of 4-aminopyridine on it.Methods Motor evoked potentials (MEPs), both spinal cord recorded MEPs (scMEPs) and extracellularly recorded MEPs (exMEPs) were recorded and characterized on a T13 epidural electrode (scMEPs) and an extracellular microelectrode (exMEPs) for 10 normal rats and 40 rats with lesions of various severity (sham, 35?g*cm force (gcf), 70?gcf, 100?gcf impact injury) at the T8-T9 cord using the Allen's drop model. The incline plane and Tarlov techniques were used to assess clinical neurological function. Results MEPs in the normal rats were elicited by applying transcortical suprathreshold stimulation consisting of 3-4 early negative peaks (N1, N2, N3 and N4) followed by several late waves. The N1 and N2 peaks were largest in the anterior and ventrolateral funiculus, respectively, which was indicative of extrapyramidal pathways. The 100?gcf impact injuries and the cord transection abolished the MEP distal to the lesion, whereas the 35?gcf injuries resulted in a latency shift and amplitude decrement of the MEP peaks. Eighteen of the 20 rats with 70?gcf injuries showed clinical paraplegia. Among them, 7 rats had neurophysiological evidence of residual conduction pathways through the lesioned cord segment, such as the presence of N1 and N2 peaks in the scMEPs or exMEPs. After 4-aminopyridine (4-AP) administrations (1?mg/kg), the amplitude of the spared exMEP increased significantly and spread more widely. Conclusions MEPs evoked by transcortical stimulation travel mostly in the extrapyramidal tract. MEP monitoring could provide an excellent method of detecting the functional integrity of the motor tracts after SCI, and could even detect spared motor fibers after discomplete SCI. Furthermore, the use of 4-AP or other K+ channel blocking agents may be a potential treatment for patients with chronic moderate to severe SCI.

Glucosamine reduces blood-brain barrier disruption by inhibiting the expression of matrix metalloproteinase-9 in experimental autoimmune encephalomyelitis rats

We investigated the effects of glucosamine(GS) on blood-brain barrier(BBB) function and matrix metalloproteinase-9 (MMP-9) expression in rats with experimental autoimmune encephalomyelitis(EAE).Animals were randomly divided into three groups,among which the EAE and GS groups were immunized with complete antigen and pertussis toxin,and the adjuvant group was immunized with complete Freund's adjuvant and pertussis toxin.Rats were treated by peritoneal injection of GS 180 mg/(kg·d) in the GS group and peritoneal injection of phosphate-buffered saline 4.5 mL/(kg·d) in the EAE and adjuvant groups.We proposed to assess the integrity of BBB by calculating cerebrospinal fluid to serum albumin quotient(QA) on days 6,8,10,12,14,16 and 18 post-immunization.At the same time,the brains and spinal cords were removed for MMP-9 immunohistochemical staining. Experiments demonstrated that in the EAE group,QA value and MMP-9 expression were highly elevated and up-regulated and correlated to disease severity.Moreover,there was statistically significantly positive correlation between QA value and MMP-9 expression.In the GS group,we observed that the mean disease onset date was delayed,the incidence and mean score of symptom were suppressed at the peak phase of disease(P<0.05).Furthermore,QA value and MMP-9 expression in the GS group showed stronger inhibition when compared with those of the EAE group(P<0.05).Our study showed that GS would reduce the BBB breakdown and leukocyte trafficking by inhibiting the production of MMP-9 and mitigate EAE.

Origin of new genes after zygotic genome activation in vertebrate

New genes are drivers of evolutionary innovation and phenotypic evolution. Expression of new genes in early development raises the possibility that new genes could originate and be recruited for functions in embryonic development, but this remains undocu- mented. Here, based on temporal gene expression at different developmental stages in Xenopus tropicolis, we found that young protein-coding genes were significantly enriched for expression in developmental stages occurring after the midblastula trans- ition （MBT）, and displayed a decreasing trend in abundance in the subsequent stages after MBT. To complement the finding, we demonstrate essential functional attributes of a young orphan gene, named as Fog2, in morphological development. Our data indicate that new genes could originate after MBT and be recruited for functions in embryonic development, and thus provide insights for better understanding of the origin, evolution, and function of new genes.