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Cheng Huang

Publications and source records attributed to Cheng Huang.

48 records · Page 3Linked to original sources

Masking of the contribution of V protein to Sendai virus pathogenesis in an infection model with a highly virulent field isolate.

Sendai virus V protein is not essential for virus replication in cultured cells but is essential for efficient virus replication and pathogenesis in mice, indicating that the V protein has a luxury function to facilitate virus propagation in mice. This was discovered in the Z strain, an egg-adapted avirulent laboratory strain. In the present study, we reexamined the function of Sendai virus V protein by generating a V-knockout Sendai virus derived from the Hamamatsu strain, a virulent field isolate, which is an appropriate model for studying the natural course of Sendai virus infection in mice. We unexpectedly found that the V-knockout virus propagated efficiently in mice and was as virulent as the wild-type virus. Switching of the functionally important V unique region demonstrated that this region of the Hamamatsu strain was also functional in a Z strain background. It thus appears that the V protein is nonsense in a field isolate of Sendai virus. However, the V protein was required for virus growth and pathogenesis of the Hamamatsu strain in mice when the virulence of the virus was attenuated by introducing mutations that had been found in an egg-adapted, avirulent virus. The V protein therefore seems to be potentially functional in the highly virulent Hamamatsu strain and to be prominent if virus replication is restricted.

Animals↗

RETRACTED: Reelin promotes peripheral synapse elimination and maturation.

Reelin is an extracellular protein that is crucial for layer formation in the embryonic brain. Here, we demonstrate that Reelin functions postnatally to regulate the development of the neuromuscular junction. Reelin is required for motor end-plate maturation and proper nerve-muscle connectivity, and it directly promotes synapse elimination. Unlike layer formation, neuromuscular junction development requires a function of Reelin that is not mediated by Disabled1 or very-low-density lipoprotein receptors and apolipoprotein E receptor 2 receptors but by a distinct mechanism involving its protease activity.

Action Potentials↗

Chromatin remodeling and human disease.

In the past few years, there has been a nascent convergence of scientific understanding of inherited human diseases with epigenetics. Identified epigenetic processes involved in human disease include covalent DNA modifications, covalent histone modifications, and histone relocation. Each of these processes influences chromatin structure and thereby regulates gene expression and DNA methylation, replication, recombination, and repair. The importance of these processes for nearly all aspects of normal growth and development is illustrated by the array of multi-system disorders and neoplasias caused by their dysregulation.

Acetylation↗

Expression of interleukin-6 mRNA in ischemic rat brain after electroacupuncture stimulation.

Interleukin-6 (IL-6) is a potent, pleiotropic cytokine that plays a central role in host defense and acute inflammatory responses, exhibiting pro- and anti-inflammatory activities. However, little is known about the effect of acupuncture on IL-6 in inflammatory responses caused by cerebral ischemia-reperfusion. The objective of present study was to investigate the influence of electroacupuncture (EA) on IL-6 mRNA expression in ischemic rat brain, by means of reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization. Data showed that EA stimulation significantly upregulated IL-6 mRNA levels in rat cortex and striatum following cerebral ischmia-reperfusion (P<0.05 vs. cerebral ischemia group, respectively), but nothing for sham-EA treatment. The result suggests that EA may partially participate in the regulation of inflammatory processes, and the mechanism of neuroprotective action of EA is implicated in modulation of IL-6 gene expression in cerebral ischemic injury.

Animals↗

Virulence of water-induced coccoid Helicobacter pylori and its experimental infection in mice.

AIM: To explore the virulence and the infectivity of coccoid Helicobacter pylori (H. pylori) transformed from spiral form by exposure to sterile tap water. METHODS: Three strains of H. pylori, isolated from gastric biopsy specimens of confirmed peptic ulcer, were converted from spiral into coccoid form by exposure to sterile tap water. Both spiral and coccoid forms of H. pylori were tested for the urease activity, and the adherence to Hep-2 cells. The presence of flagella was examined under electron microscopy. In the experimental animal infection, the spiral and coccoid forms of H. pylori originated from the same strain F49 were inoculated intragastrically into BALB/c mice respectively four times at a 3-day interval. Half of the mice from each group were sacrificed at Day 21 and Day 28 after the last inoculation. Histology and H. pylori colonization were detected by urease test of gastric mucosa, cultures of H. pylori, and electron microscopy and so on. RESULTS: The urease activity and the ability of adherence to Hep-2 cells were found to be lower in coccoid H. pylori than that in its spiral form. For example, the transformation in strain F(44) led to a significant decrease of the adherence rate and adherence index from 70.0+/-5.3 % to 30.2+/-3.5 % (P<0.01), and from 2.6+/-0.4 to 0.86+/-0.3 (P<0.01), respectively. The flagella of coccoid H. pylori were observed under electron microscope. In the experimental infection in mice, the positive rate of gastric mucosa urease test was 93.8 % (15/16) in the group infected by spiral H. pylori and 50 % (8/16) in the group infected by coccoid H. pylori, and the estimated coccoid H. pylori colony number was 1.75 vs 0.56. The positive rates of H. pylori culture were 87.5 % (14/16) in spiral H. pylori group and 68.8 % (11/16) in coccoid H. pylori group. There was no significant difference in either urease test or bacterial culture rate between the groups examined at Day 21 and Day 28 after inoculation. Electron microscopic examination of the samples taken from both groups showed the adherence of H. pylori in spiral, bacillary and coccoid shapes to the epithelial cells of gastric wall. Histological examination showed the occurrence of gastric mucosal injury as indicated by various degrees of erosion, ulcer, and inflammatory cell infiltration. Mucosal injury was slighter in the mice infected by coccoid H. pylori. No positive result was obtained in the control group that received intragastrical administration of sterile tap water. CONCLUSION: Although the virulence of coccoid H. pylori induced by water decrease, coccoid H. pylori still remains a considerable urease activity and the adhering ability to epithelial cells. Furthermore, the flagella, an important component responsible for bacterial movement and infection, were still observed as a cellular structure of coccoid H. pylori under electron microscope. The coccoid H. pylori induced by water is capable of colonizing in gastric mucosa and causing gastrititis in mice.

Animals↗

An all-organic composite actuator material with a high dielectric constant.

Electroactive polymers (EAPs) can behave as actuators, changing their shape in response to electrical stimulation. EAPs that are controlled by external electric fields--referred to here as field-type EAPs--include ferroelectric polymers, electrostrictive polymers, dielectric elastomers and liquid crystal polymers. Field-type EAPs can exhibit fast response speeds, low hysteresis and strain levels far above those of traditional piezoelectric materials, with elastic energy densities even higher than those of piezoceramics. However, these polymers also require a high field (>70 V micro m(-1)) to generate such high elastic energy densities (>0.1 J cm(-3); refs 4, 5, 9, 10). Here we report a new class of all-organic field-type EAP composites, which can exhibit high elastic energy densities induced by an electric field of only 13 V micro m(-1). The composites are fabricated from an organic filler material possessing very high dielectric constant dispersed in an electrostrictive polymer matrix. The composites can exhibit high net dielectric constants while retaining the flexibility of the matrix. These all-organic actuators could find applications as artificial muscles, 'smart skins' for drag reduction, and in microfluidic systems for drug delivery.

Artificial Organs↗

Mutational analysis of the Sendai virus V protein: importance of the conserved residues for Zn binding, virus pathogenesis, and efficient RNA editing.

The V protein of Sendai virus (SeV) is nonessential for virus replication in cell culture but indispensable for viral pathogenicity in mice. At the C terminus of the V protein, there are amino acid residues conserved among the members of the Paramyxovinae subfamily that are clustered in three regions: region I, just downstream of the RNA editing site; and regions II and III, cysteine-rich zinc-finger-like regions. In the present study, we introduced mutations into the conserved amino acids and generated nine mutant viruses. All of the viruses had impaired virus replication in mouse lungs and attenuated virulence in mice. Furthermore, the C-terminal polypeptides fused with glutathione-S-transferase with a mutation in region I, II, or III all had impaired Zn binding in a (65)Zn-binding assay in solution. These results demonstrate that the conserved amino acids are important for V protein function, probably via protein conformation dependent on Zn binding. One mutant, SeV V-H(318)N, had inefficient RNA editing, indicating that the nucleotide that is a part of the codon encoding histidine at position 318 is conserved for the RNA editing machinery. In addition, to determine the function of the C-terminal extension of the V protein, which is not translated in recent virulent field isolates, a translational stop codon was introduced to generate the corresponding short V protein. The mutant virus showed similar virus propagation and pathogenicity, indicating that C-terminal extension of the V protein is not relevant to virus pathogenesis.

Amino Acid Sequence↗

Characteristics of electroacupuncture-induced analgesia in mice: variation with strain, frequency, intensity and opioid involvement.

The present study was conducted to evaluate the characteristics of electroacupuncture (EA)-induced analgesia in mice. Three inbred strains of mice (DBA/2, C57BL/6J, BALB/c) and three outbred strains (ICR, LACA, NIH) were used in the experiment. Two pairs of metallic needles were inserted into acupoints ST 36 and SP 6 connected to an electric pulse generator. EA parameters were set as constant current output with alteration of a positive and negative square wave, 0.6 ms in pulse width for 2 Hz and 0.3 ms for 100 Hz. Tail-flick latencies evoked by radiant heat were measured before, during and after EA stimulation. We found that (1) DBA/2 mice showed a significantly more potent analgesic effect than the other five strains in response to both 100 and 2 Hz EA. In this case, the intensities were 1.0-2.0-2.0 mA, 10 min for each intensity totally 30 min. (2) EA analgesia increased as the intensity of stimulation increased from 0.5 to 2.0 mA, but it remained at this plateau when the intensity further increased from 2.0 to 3.0 mA. (3) 10.0 mg x kg(-1) naloxone was needed to block the analgesic effect induced by 2 Hz EA of 2.0 mA, but to block that by 100 Hz, 25.0 mg x kg(-1) was necessary. (4) A positive correlation was observed between analgesia induced by morphine at the dose of 5.0 mg x kg(-1) and by 100 Hz EA in two tested strains DBA/2 and C57BL/6J. In conclusion, EA induces reliable, strain-dependent analgesia in mice. The naloxone-reversibility of EA, a measure of whether it is opioid or non-opioid mediated, is dependent upon intensity and frequency.

Acupuncture Analgesia↗

Involvement of the leader sequence in Sendai virus pathogenesis revealed by recovery of a pathogenic field isolate from cDNA.

We previously demonstrated that a systematic passage of a pathogenic field isolate of Sendai virus (SeV), the Hamamatsu strain, in embryonated eggs caused attenuation of virulence to mice, and we isolated viral clones of distinct virulence (K. Kiyotani et al. Arch. Virol. 146:893-908, 2001). One of the clones, E15cl2, which was obtained from the virus at the 15th egg passage of E0, the parental Hamamatsu clone for egg passage, had 165-fold-attenuated virulence to mice and possessed only four mutations in the entire 15,384-base genome: in an antigenomic sense, U to A at position 20 (U20A) and U to A at position 24 (U24A) in the leader sequence, the promoter for transcription and replication, and A to G at position 9346 (silent) and A to U at position 12174 (Ser to Cys) in the L gene. To examine the possibility that leader mutations affect virus pathogenesis, we recovered live viruses from cDNA derived from the Hamamatsu strain. A mutant virus possessing either a mutation of U20A or U24A in the leader sequence showed a slightly lower pathogenicity than that of the parental virus, whereas a double mutant virus possessing both of the mutations showed 25-fold-attenuated virulence, accompanying a significantly lower virus replication in the mouse lung. Replications of the leader mutant viruses were also impaired in a primary culture of mouse pulmonary epithelial cells but not in chicken embryo fibroblasts. These findings suggest that leader mutations of SeV affect virus pathogenesis by altering virus replication in a host-dependent manner.

5' Untranslated Regions↗

Alteration of Sendai virus morphogenesis and nucleocapsid incorporation due to mutation of cysteine residues of the matrix protein.

The matrix (M) protein of Sendai virus (SeV) has five cysteine residues, at positions 83, 106, 158, 251, and 295. To determine the roles of the cysteine residues in viral assembly, we generated mutant M cDNA possessing a substitution to serine at one of the cysteine residues or at all of the cysteine residues. Some mutant M proteins were unstable when expressed in cultured cells, suggesting that cysteine residues affect protein stability, probably by disrupting the proper conformation. In an attempt to generate virus from cDNA, SeV M-C(83)S, SeV M-C(106)S, and SeV M-C(295)S were successfully recovered from cDNA, while recombinant SeVs possessing other mutations were not. SeV M-C(83)S and SeV M-C(106)S had smaller virus particles than did the wild-type SeV, whereas SeV M-C(295)S had larger and heterogeneously sized particles. Furthermore, SeV M-C(106)S had a significant amount of empty particles lacking nucleocapsids. These results indicate that a single-point mutation at a cysteine residue of the M protein affects virus morphology and nucleocapsid incorporation, showing direct involvement of the M protein in SeV assembly. Cysteine-dependent conformation of the M protein was not due to disulfide bond formation, since the cysteines were shown to be free throughout the viral life cycle.

Animals↗

Study of the mechanisms of acupuncture and moxibustion treatment for ulcerative colitis rats in view of the gene expression of cytokines.

AIM:To observe the effect of acupuncture and moxibustion on the expression of IL-1beta and IL-6 mRNA in ulcerative colitis rats.METHODS:The SD rat ulcerative colitis model was created by immunological method associated with local stimulation. Colonic mucosa was prepared from human fresh surgical colonic specimens, homogenized by adding appropriate amount of normal saline and centrifuged at 3000r/min. The supernatant was collected for measurement of protein conentration and then mixed with Freund adjuvant. This antigen fluid was first injected into the plantae of the model group rats, and then into their plantae, dorsa, inguina and abdominal cavities (noFreund adjuvant for the last injection) again on the 10th, 17th, 24th and 31st day. When a certain titer of serum anti colonic antibody was reached, 2% formalin and antigen fluid (no Freund adjuvant) were administered separately by enema. The ulcerative colitis rat model was thus set up. The animals were randomly divided into four groups: model control group (MC, n = 8), electro acupuncture group (EA, n = 8), herbs partition moxibustion group (HPM 8), normal control group (NC,n = 8). HPM: Moxa cones made of refined mugwort floss were placed on the medicinal pad (medicinal pad dispensing: Radix Aconiti praeparata, cortex Cinnamomi, etc) for Qihai (RN 6) and Tianshu (ST 25, bilateral) and ignited. Two moxa cones were used for each acupoint once a day and 14 times in all. EA: Tianshu (bilateral) and Qihai were stimulated by the intermittent pulse with 2Hz frequency, 4mA intensity for 20 minutes once a day and 14 times in all. After treatment, rats of all four groups were killed simultaneously. The spleen was separated and the distal colon was dissected. Total tissue RNA was isolated by the guanidinium thiocyanate phenol chloroform extraction method. RT-PCR technique was used to study the expression of IL-1 beta and IL-6 mRNA.RESULTS:IL-1 beta and IL-6 mRNAs were not detected in the spleen and colonic mucosa of the NC rats, whereas they were significantly expressed in that of the MC rats.IL-1 beta and IL-6 mRNAs were markedly lower in the EA and HPM rats than that in MC rats. There was no significant difference between the levels of IL-1 beta and IL-6 mRNAs in the EA and HPM rats. The expressions of IL-1 beta and IL-6 mRNAs were nearly the same in the spleen and colon of all groups.CONCLUSION:Acupuncture and moxibustion greatly inhibited the expression of IL-1 beta and IL-6 mRNA in the experimental ulcerative colitis rats.

Journal Article↗