Design and expression of FGF23 inhibitor in Pichia pastoris / 药学学报

Hypophosphatemia is a common metabolism disease in humans. Fibroblast growth factor 23 (FGF23) inhibits phosphate reabsorption by targeting on the renal tubules. FGF23C-tail contains 73 amino acids from C-terminus of FGF23, serves as an inhibitor of FGF23, and can increase phosphate reabsorption. Therefore, FGF23C-tail is an important drug for hypophosphatemia. In this paper, we constructed a fusion protein of FGF23C-tail with HSA, and investigated the expression of the fusion protein in the Pichia pastoris system. The recombinant gene was constructed by fusion PCR. A high-yield strain was selected by G418 resistance and fermentation yield, and the expression yield was 43.7 mg·L-1 in flask. In 5 L fermenters, the highest expression yield could reach 265.6 mg·L-1. FGF23C-tail-HSA could be used as an inhibitor for FGF23, and could significantly increase blood phosphorus levels in rats. The procedures for care and use of animals were approved by the Ethics Committee of YiChun University. This paper provided a basis research for further studying physiological activity of FGF23C-tail-HSA.

Research progress on non-injection administration systems of paclitaxel / 中草药

Paclitaxel (PTX) is a complex secondary metabolite isolated from Taxus brevifolia, which widely used as chemotherapentic agent with a broad spectrum of actinity against cancer in the world. Its water solubility was poor and oral bioavailability was low. Cremophor-EL was used in traditional PTX injections to improve the solubility of PTX, and resulted in several adverse side effects such as severe hypersensitivity. Pre-desensitization treatment was needed before clinical use. Recently, a variety of non-injection drug delivery systems (DDS) of PTX have been developed. In this paper, the research progress of non-parenteral PTX was reviewed, including oral administration systems, vaginal administration systems, transdermal DDS, implantable DDS, transdermal DDS, intranasal administration and inhalation DDS, so as to provide references for future study and clinical applications.

Complete genomic analysis of a novel infectious bronchitis virus isolate / 病毒学报

The genome of CK/CH/SD09/005, an isolate of infectious bronchitis virus (IBV), was characterized to enable the further understanding of the epidemiology and evolution of IBV in China. Twenty-five pairs of primers were designed to amplify the full-length genome of CK/CH/SD09/005. The nucleotide sequence of CK/CH/SD09/005 was compared with reference IBV strains retrieved from GenBank. The phylogenic relationship between CK/CH/SD09/005 and the reference strains was analyzed based on S1 gene sequences. The complete genome of CK/CH/SD09/005 consisted of 27691 nucleotides (nt), excluding the 5' cap and 3' poly A tail. The whole-genome of CK/CH/SD09/005 shared 97 - 99% nucleotide sequence homology with the GX-NN09032 strain, which was the only complete genome that was closely related to CK/CH/SD09/005. When compared with all reference strains except GX-NN09032, CK/CH/SD09/005 showed the highest similarity to ck/CH/LDL/091022 and SDIB821/2012 (QX-like) in the replicase gene (Gene 1) and 3'UTR, with a sequence identity rate of 97% and 98%, respectively. However, CK/CH/SD09/005 exhibited lower levels of similarity with ck/CH/LDL/091022 and SDIB821/2012 in S-3a-3b-3c/ E-M-5a-5b-N with a sequence identity of 72% - 90%. CK/CH/SD09/005 showed the highest level of nucleotide identity with Korean strain 1011, and Chinese strains CK/CH/LXJ/02I, DK/CH/HN/ZZ2004 and YX10, in ORF 3c/E (97%), 5a (96%), 5b (99%) and N (96%), respectively. ORFs 3a, 3b and M of CK/CH/SD09/005 exhibited no more than 90% homology with the reference strains, excluding GX-NN09032. The phylogenic analysis based on the S1 gene revealed that CK/CH/SD09/005 and 39 published strains were classified into seven clades (genotypes). CK/CH/SD09/005 was distributed in clade IV with several isolates collected between 2007 and 2012. CK/CH/SD09/005 showed 66% - 69% and 72% - 81% nucleotide identities with the IBV strains of other six clades in the S1 and S2 subunits, respectively. More over, multiple substitutions were found throughout the entire S gene of CK/CH/SD09/005, while insertions and deletions were located within the S1 gene. These results indicated that CK/CH/SD09/005 is a novel variant that may be derived from the QX-like strains that are prevalent in China. Multiple genetic mechanisms, including recombinations, mutations, insertions and deletions, are likely to have contributed to the emergence of this IBV strain.

Cited status and its correlated factors of articles published in Chinese Journal of Pediatrics from 2001 to 2010 / 中华儿科杂志

<p><b>OBJECTIVE</b>The present study was designed to explore the cited status and its correlated factors of articles published in Chinese Journal of Pediatrics.</p><p><b>METHOD</b>Articles published in Chinese Journal of Pediatrics from 2001 to 2010 were searched using Wanfang Medical Online database, and the relationship between cited number and column and funding status were analyzed.</p><p><b>RESULTS</b>Totally 3209 articles were published by Chinese Journal of Pediatrics from 2001 to 2010. Two thousand and seventy-three articles (64.60%) were cited. The total cited number was 18 546 (mean rate: 5.78 per paper). Standard/protocol/guideline was the most often cited column (mean rate: 62.92 per paper). Featured articles including editorials (mean rate: 7.12 per paper), special articles (mean rate: 6.50 per paper) and original articles (mean rate: 7.90 per paper) had higher cited rate. Cited rate of original papers in featured articles was higher than other original articles (mean rate: 6.03 per paper). Those with academic perspectives, such as Opinion/Debate/Discussion, were well cited (mean rate: 7.09 per paper). All of original articles in Rapid Pathway were well cited (mean rate: 16.20 per paper). Mean cited number of grant-supported articles was 4.81 per paper, lower than that of all kinds of papers (mean rate: 5.78 per paper) and non-grant-supported articles (mean rate: 6.06 per paper). However, it was slightly higher than that of articles except Standard/protocol/guideline (mean rate: 4.36 per paper).</p><p><b>CONCLUSION</b>Cited status varies among columns. The invited papers should be increased in number to raise commentary papers and original articles with high quality.Grant-supported or non-grant-supported papers should be reviewed based on the same standard after submission.</p>

Biological characteristics of three Newcastle disease virus isolates and entire genome sequences analysis / 病毒学报

Three Newcastle disease virus (NDV) strains recovered from ND outbreaks in chickens and duck flocks in north china during 2009 to 2011 were completely sequenced and biologically characterized. All the strains were velogenic and had the velogenic motif 112R-R-Q-K-R-F117 which was consistent with the results of biological tests. Analysis of the variable region (nucleotide 47 to 420) of the F gene indicated that the three isolates belonged to genotype VII d. Cross hemagglutination inhibition test indicated that the antigen homology between three isolates and LaSota were 82.5%-89.4%, the homology between the two isolates from chicken was 90%. A cross-protection experiment in which specific-pathogen-free chickens vaccinated with LaSota were challenged by SDLY01 isolate showed that LaSota vaccine could provide complete protection against SDLY01, however virus discharge could be detected on fifth day. Challenge experiment in which Cherry Valley duck of 30 day old challenged with SD03 strain indicated that cherry valley duck had no disease in experiment period, but virus discharge could be detected from Larynx and cloaca until fifth day. Genome length of three NDV isolates was 15192bp and belonged to genotype VII d. Sequence analysis clarified that the whole genomic sequence of these three isolates shared high homology with NDV virus strains isolated from goose and duck over the same period, which elucidated that NDV isolated from goose, duck or chicken had close genetics and epidemiological relationship.

Clinical effect of sphenopalatine ganglion needling in treating primary trigeminal neuralgia of Liver-yang upsurge syndrome type / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the efficacy and safety of sphenopalatine ganglion needling in treating patients with primary trigeminal neuralgia (PTN) of Liver (Gan)-yang upsurge syndrome (LYUS) type.</p><p><b>METHODS</b>Sixty-five PTN patients of LYUS type were assigned by a random number table to two groups and treated by deep-needling (33 patients, DN group) and superficial-needling (32 patients, SN group), respectively. The main-acupoint used for both groups was Xiaguan (ST7) of affected side, on which needle was deeply inserted to reach spheno-palatine ganglion in the DN group, but was inserted conventionally in the SN group; the auxiliary acupoints used were the local points Cuanzhu (BL2), Sibai (ST2), Chengjiang (CV24) of affected side, as well as the remote points, bilateral Hegu (LI4) and Taichong (LR3). The needling was implemented every other day, with electric stimulation applied to all the punctured acupoints for 30 min, 10 days treatment as one therapeutic course. Each patient received 2 courses of needling with 1 week interval in between. The clinical efficacy, pain control and adverse reaction were evaluated and compared after the treatment.</p><p><b>RESULTS</b>Rank-sum test showed that the efficacy in the DN group was significantly better than that in the SN group (Z =2.30, P=0.021); the pain was alleviated in both groups in frequency, intensity and sustained time (evaluated by visual analogue scale) significantly (P<0.01), but deep needling showed a superiority over superficial needling in reducing the frequency and intensity of episode (Z=9.55, Z=5.50, both P=0.00). No adverse reaction occurred during the treatment course of both groups.</p><p><b>CONCLUSION</b>Spheno-palatine ganglion needling is highly effective, safe and reliable for the treatment of PTN.</p>

Trigeminal neuralgia of hyperactive of liver yang type treated with acupuncture at Xiaguan (ST 7) at different depth: a randomized controlled trial / 中国针灸

<p><b>OBJECTIVE</b>To observe the differences of therapeutic effect in primary trigeminal neuralgia (PTN) of hyperactive of liver yang type treated by deep and shallow puncturing at Xiaguan (ST 7).</p><p><b>METHODS</b>Sixty-three cases of PTN of hyperactive of liver yang type were randomly divided into a deep puncturing group (32 cases) and a shallow puncturing group (31 cases). Xiaguan (ST 7) of affected region, Hegu (LI 4) and Taichong (LV 3) of bilateral sides, Cuanzhu (BL 2), Sibai (ST 2) and Jiachengjiang (Extra) relevant to the affected branch of nerve stem were selected in both groups. In deep puncturing group, Xiaguan (ST 7) was punctured to the depth of spheno-palatine ganglion (SPG); Cuanzhu (BL 2), Sibai (ST 2) and Jiachengjiang (Extra) were respectively punctured to the depth of supraorbital foramen, infraorbital foramen and mental foramen. In shallow group, routine puncturing was applied; the needles were connected with G6805 electric acupuncture apparatus, and switched on for 30 min every time; the treatment was applied every other day. Pain index, traditional Chinese medicine symptoms index and clinical therapeutic effect were observed after 2 courses of treatment.</p><p><b>RESULTS</b>In deep puncturing group, the VAS scores and the traditional Chinese medicine symptoms scores (pain degree, pain frequency, upsetting, conjunctival congestion, bitter mouth and hypochondriac pain) after treatment were much more lower than those before treatment (all P < 0.01); in shallow puncturing group, except hypochondriac pain (P > 0.05), other indices above after treatment were obviously lower than those before treatment (P < 0.01, P < 0.05). Compared with the indices in both groups after treatment, the VAS scores, the pain degree, conjunctival congestion and total scores of traditional Chinese medicine symptoms in deep puncturing group were more significant (all P < 0.05). The total effective rate was 93.8% (30/32) in deep puncturing group, superior to that of 87.1% (27/31) in shallow puncturing group (P < 0.05). No any adverse reaction was observed in both groups.</p><p><b>CONCLUSION</b>The therapeutic effect of trigeminal neuralgia of hyperactive of liver yang type treated with electroacupuncture is remarkable, and deep puncturing at Xiaguan(ST 7) to SPG is more effective than routine puncturing.</p>

Molecular epidemiological analysis of class I Newcastle disease virus isolated from China in 2008 / 病毒学报

Thirteen isolates of Class I Newcastle disease virus obtained from healthy poultry in China during 2008 were characterized genotypically in this study. All the isolates were proved to be lentogenic strains based on the deduced amino acid sequence of the Fusion protein gene. Molecular epidemiological analysis showed that 13 isolates could be subdivided into 2 distinct genotypes, 11 isolates belonged to genotype 2, and other 2 isolates belonged to genotype 3. Results indicated two genotypes of Class I Newcastle disease virus might widely exist in domestic poultry in China.

Paeonol attenuates oxygen-glucose deprivation injury and inhibits NMDA receptor activation of cultured rat hippocampal neurons / 药学学报

The purpose of this study is to determine if paeonol can protect hippocampal neurons against injury due to oxygen-glucose deprivation (OGD) injury. The rat neurons were cultured in an OGD environment and the model of OGD injury was established. Paeonol and MK-801, a positive control drug, were added before deprivation. Neuron viability was measured by the reduction of MTT; glutamate was analyzed by amino acid analyzer; binding activity of NMDA receptor was evaluated by liquid scintillation counting and the expression of NMDA receptor NR1 subunit mRNA was semiquantitatively determined by RT-PCR. Compared with OGD injury group, paeonol treatment obviously increased cell survival rate and reduced the binding activity of NMDA receptors and the release of glutamate; and down-regulating the expression of NR1 subunit. These results suggest that paeonol may exhibit its protective effect against OGD injury by the action on NMDA receptor of rats.

Effects of iodine excess on TPO and NIS genes mRNA expression in rats / 中国地方病学杂志

Objective To observe the effects of iodine excess on thyroid morphology,the expression of thyroid peroxidase and sodium iodide symporter mRNA and to explore their mechanisms.Methods One-month SD rats were divided into three groups:control iodine(CI),high iodine Ⅰ(HI Ⅰ)and high iodineⅡ(HI Ⅱ)and were fed with water containing iodine in different concentrations by adding K103(5,5000,10 000μg/L)respectively.Rats were sacrificed after being fed for six months.The morphology of thyroid was investigated under light microscopy and electron microscopy,the serum thyroid hormones and ratio of TPO/β-actin and NIS/β-actin were measured by radio-immunoassay and RT-PCR method.Results The major changes were increased follicles with colloid accumulation in HI groups.The levels of serum thyroid hormones TT3 and TT4 were decreased gradually from CI[(75.68±13.99,1.45±0.49)nmol/L]to HI Ⅰ[(73.82±16.48,1.34±0.31)nmol/L]and HIⅡ groups[(70.65±11.43,1.15±0.39)nmol/L],but there were no significant differences among three groups(F=O.371,l.163,P＞0.05).The TPO and NIS mRNA expressions in HI Ⅰ(1.28±0.10,0.56±O.17)and HI Ⅱ(1.14±0.04,0.39±0.06)were significantly lower(F=30.863,62.62.675,P＜O.05)than those of control group(1.39±0.08,0.71±0.13).Conclusions Chronic iodine excess leads to definite histological changes in rat thyroid,and inhibits the expressions of TPO and NIS mRNA as well as thyroid hormone synthesis,which in turn acts as a protective mechanism against iodine excess.

Hydroxyethylpuerarin attenuates focal cerebral ischemia-reperfusion injury in rats by decreasing TNF-alpha expression and NF-kappaB activity / 药学学报

This study is to investigate the effect of hydroxyethylpuerarin on the expression of tumor necrosis factor-alpha (TNF-alpha) and activity of nuclear factor kappa B (NF-kappaB) after middle cerebral artery occlusion (MCAO) in rats. Rats were subjected to cerebral ischemia-reperfusion injury induced by MCAO. Hydroxyethylpuerarin (10, 20, 40 mg x kg(-1), iv) was administered just 30 min before occlusion and immediately after reperfusion. After a 24 h reperfusion following 2 h of MCAO, the number of viable neurons in hippocampal CA1 region was counted by hematoxylin and eosin (HE) staining. TNF-alpha protein and its mRNA expression were examined with radioimmunoassay (RIA) and reverse transcriptasepolymerase chain reaction (RT-PCR) respectively. NF-KB activity was observed by electrophoretic mobility shift assay (EMSA), and inhibition of NF-kappaB alpha (IkappaBalpha) protein expression was evaluated by Western blotting analysis. Animals treated with hydroxyethylpuerarin had a significant increase in neuronal survival in comparison with vehicle-treated group. Hydroxyethylpuerarin significantly reduced the protein and mRNA expression of TNF-alpha following 2 h of ischemia with 24 h of reperfusion. NF-kappaB DNA binding activity and the degradation of IkappaBalpha in the cytoplasm also decreased by hydroxyethylpuerarin treatment. The protective effects of hydroxyethylpuerarin against ischemia-reperfusion injury may be mediated by decreasing the expression of TNF-alpha and the activity of NF-kappaB in rats.

Effect of compound EXP-2528 on angiotensin II-induced E-selectin and VCAM-1 expression in rat brain microvascular endothelial cells in vitro / 药学学报

The aim of this study is to investigate the effect and mechanism of angiotensin (Ang) II on E-selectin and vascular cell adhesion molecule-1 (VCAM-1) expression in rat brain microvascular endothelial cells (BMEC) and evaluate the effect of compound EXP-2528, a novel Ang II type 1 (AT1) receptor antagonist. The experiment was performed in cultured BMEC of rat. The mRNA and protein expression of E-selectin and VCAM-1 in BMEC was analyzed by RT-PCR and Western blotting, respectively. The results showed that the mRNA and protein expression of E-selectin and VCAM-1 in BMEC were significantly upregulated by 4 h or 18 h exposure to 1 x 10(-7) mol x L(-1) Ang II. These effects were abolished by pretreatment with the selective AT1 receptor antagonists losartan and compound EXP-2528, but not with the AT2 selective antagonist PD123319. Combining losartan with PD123319 also significantly inhibited Ang II-induced E-selectin and VCAM-1 expression in BMEC, but there was no significant difference compared with losartan group. These findings indicated that Ang II upregulated E-selectin and VCAM-1 in BMEC by activating AT1 receptor and then involved in the development of cerebrovascular disease.

Effect of baicalin on HSP70 expression of hippocampal neurons in focal brain ischemia-reperfusion injury rats / 药学学报

<p><b>AIM</b>To investigate the effect of baicalin on the hippocampal neuronal apoptosis and the expression of HSP70 in rats with focal brain ischemia-reperfusion injury.</p><p><b>METHODS</b>One hundred and twenty male Wistar rats were randomly divided into six groups:sham operated group, ischemia-reperfusion group, nimodipine group and three baicalin groups,to which baicalin was administered at doses of 50, 100 and 200 mg x kg(-1), separately. The models of focal brain ischemia-reperfusion injury induced by middle cerebral artery occlusion (MCAO) were used in this study. HE stain was used to observe the pathological changes. Flow cytometry (FCM) was used for determination of neuronal apoptosis. HSP70 protein expression of the neurons was detected with immunohistochemistry. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of the mRNA level of HSP70.</p><p><b>RESULTS</b>Baicalin can significantly relieve the pathological changes and inhibit apoptosis in hippocampus CA1 area, and at the same time increase the expression of HSP70 and HSP70 mRNA.</p><p><b>CONCLUSION</b>Baicalin can relieve brain damage induced by focal brain ischemia-reperfusion in rats, which may be related to inhibiting the process of the neuronal apoptosis. The mechanism of antiapoptosis effect of baicalin may be related to the promotion of transcription of HSP70 mRNA and increasing the expression of the protein.</p>

Protective effects of hydroxyethylpuerarin against brain astrocytes injury induced by hydrogen peroxide / 药学学报

<p><b>AIM</b>To study the protective effects of hydroxyethylpuerarin against the injury of astrocytes induced by hydrogen peroxide (H2O2).</p><p><b>METHODS</b>Experiments were performed with cells from passage 4. Plasma membrane integrity was measured by lactate dehydrogenase (LDH) release. The occurrence of apoptosis was measured by flow cytometry. The glutamate uptake of astrocytes was studied with [3H]-glutamate incorporation. Intracellular superoxide dismutase (SOD) activity and malondialdehyde (MDA) level were assessed by automatic biochemistry analyzer.</p><p><b>RESULTS</b>Compared with H2O2 injured group, the occurrence of apoptosis, levels of LDH release and intracellular MDA of astrocytes reduced in hydroxyethylpuerarin pre-treated groups, but the glutamate uptake and intracellular SOD activity of astrocytes increased.</p><p><b>CONCLUSION</b>Hydroxyethylpuerarin could reduce the occurrence of apoptosis and improve neurotrophic function of astrocytes, which may be related with its antioxidant effects during oxidative stress.</p>

Protective effects of hydroxyethylpuerarin on cultured bovine cerebral microvascular endothelial cells damaged by hydrogen peroxide / 药学学报

<p><b>AIM</b>To observe the damages induced by hydrogen peroxide in cultured bovine cerebral microvascular endothelial cells (BCMEC) and evaluate the protective effects of hydroxyethylpuerarin on hydrogen peroxide-injured BCMEC.</p><p><b>METHODS</b>BCMEC were cultured and transferred into modified Eagle medium (MEM). The viability of cells was detected by MTT assay. Cell injury was determined by lactate dehydrogenase (LDH) activity in the extracellular medium. Flow cytometry was employed to observe the occurrence of apoptosis. Morphologic changes of cells were visualized under phase contrast and electron microscopes.</p><p><b>RESULTS</b>Hydrogen peroxide (200 micromol x L(-1) for 4 hours) inhibited the viability of cultured BCMEC and stimulated LDH release. Hydrogen peroxide (100 micromol x L(-1) for 4 hours) induced the occurrence of apoptosis. Hydroxyethylpuerarin was shown to increase the survival rate and decrease the activity of LDH of BCMEC damaged by hydrogen peroxide. Hydroxyethylpuerarin was also found to protect BCMEC against apoptosis induced by hydrogen peroxide.</p><p><b>CONCLUSION</b>Hydrogen peroxide induces BCMEC injury either by apoptosis or through necrosis. Hydroxyethylpuerarin protects BCMEC against hydrogen peroxide-induced injury in a concentration-dependent manner. Its antioxidant effects might be involved as the mechanism protection.</p>

Potential effect of tumor necrosis factor-alpha and its receptor II gene polymorphisms on the pathogenesis of coal worker's pneumoconiosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To approach the effect of tumor necrosis factor-alpha (TNF-alpha) and tumor necrosis factor receptor II (TNFRII) gene polymorphisms on genetic susceptibility of coal worker's pneumoconiosis and their relationship with pulmonary fibrosis.</p><p><b>METHODS</b>Two hundred and thirty-four cases of coal worker's pneumoconiosis (CWP) and four hundred and forty coal mine workers (controls) were selected, and the cases of CWP were divided into three subgroups based on the various stages of I, II and III. 3 ml peripheral vein blood was drawn from every subject. Using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) techniques, TNF-alpha and TNFRII gene polymorphisms were analyzed.</p><p><b>RESULTS</b>In both group matching and 1:1 paired matching, there was no significant difference between CWP workers and controls in distribution frequencies of G/A + A/A (TNF-alpha -308) and T/G + G/G (TNFRII 196) genotypes. The distribution frequency of G/A + A/A genotype in CWP with stage III (20.00%) was higher than those in control (10.91%), and CWP cases with stage I (10.34%) and II (7.50%) respectively. The risk of CWP with stage III in those with G/A + A/A genotype was 2-fold higher than with G/G genotype (OR = 3.00, 95% CI: 0.35 approximately 25.84) for 1:1 paired matching.</p><p><b>CONCLUSIONS</b>TNF-alpha and TNFRII gene polymorphisms does not play an important role in susceptibility to CWP of Han race. TNF-alpha gene promoter polymorphisms might be related with the degree of severe pulmonary fibrosis in CWP.</p>