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1.
Indian J Pathol Microbiol ; 2022 Mar; 65(1): 208-210
Article | IMSEAR | ID: sea-223208

ABSTRACT

Desmoplastic small round cell tumor (DSRCT) is a very rare diagnosis with about 200 cases reported in literature. DSRCT is a recently described histopathological entity by Gerald and Rosai in 1989. Abdominopelvic cavity especially peritoneum is the most common site. We report a case of a huge omental DSRCT with lymph node metastasis which was initially misdiagnosed as gastrointestinal stromal tumor on radiology. A 26-year-old male presented with complaints of upper abdominal swelling associated with constant dull pain. On examination there was a large 15 × 12 cm intraabdominal mass in the epigastric and umbilical region. Imaging studies were suggestive of neoplastic mesenchymal etiology. Image-guided fine-needle aspiration cytology (FNAC) was suggestive of mesenchymal neoplastic etiology. On laparotomy, there was a huge 20 × 15 cm mass arising from omentum with multiple omental and mesenteric seedlings and mesenteric, peripancreatic and perigastric lymphadenopathy. The patient underwent debulking surgery with uneventful post-operative recovery. Histopathological examination with immunohistochemistry revealed a diagnosis of DSRCT of omentum and small bowel mesentery with lymph node metastasis. Patient then received adjuvant chemotherapy with multiple chemotherapeutic drugs as per P6 protocol and has stable disease at 1 year follow up.

2.
Chinese Journal of Tissue Engineering Research ; (53): 4012-4017, 2020.
Article in Chinese | WPRIM | ID: wpr-847324

ABSTRACT

BACKGROUND: Corneal epithelial stem cells, also known as limbal stem cells, are distributed in the basal layer of limbal epithelium. It is extremely difficult to deal with limbal stem cell deficiency or dysfunction that is caused by severe thermal burn, chemical burn, and chronic inflammation of ocular surface. At present, in vitro culture of corneal epithelial stem cells using tissue engineering technology followed by clinical transplantation is a new and effective therapeutic direction. OBJECTIVE: To explore the feasibility of serum-free culture of human corneal epithelial stem cells in vitro using modified explant culture method. METHODS: The remaining donor corneal tissues after keratoplasty (less than 8 mm in diameter) were obtained from Henan Eye Bank, and the outer and middle limbus were dissected under surgical microscope. Two culture methods were used to culture human corneal epithelial stem cells. In the conventional explant culture group, the limbal tissues were adhered to the dish with the epithelium being upward, then Keratinocyte-serum free medium (K-SFM) was added into dishes, followed by incubation at 37 °C in a 5% CO2 incubator. In the modified explant culture group, limbal tissues were dissected to immerse in the K-SFM culture medium and incubated at 37 °C in the 5% CO2 incubator for 12 hours. The limbal tissues were then adhered to the dish with the epithelium being downward. The day whenever the cells from the limbal tissues adhered to the dish was marked as the 1st day of culture, and changes in cell morphology and growth were recorded by phase contrast microscopy every day. Immunofluorescent staining was used to detect the expression of K3 and p63 in primary cells on the 5th, 10th and 14th day of the modified explant culture. RESULTS AND CONCLUSION: The mean early stage of growth in the modified explant culture group was shorter than that of the conventional explant culture group (P < 0.05), and the mean growth rate of the modified explant culture group was higher than that of the conventional explant culture group (P < 0.05). In the modified explant culture group, cells had a good growth state, and many cells with small size gathered together on the 10th day of culture. On the 14th day, cell clones were formed, and the cells in the clone showed uniform morphology. On the 5th day, K3 highly expressed, while p63 lowly expressed in primary cells. On the 10th day, both of K3 and p63 had an increased expression. On the 14th day, there was no significant increase in the K3 expression, but the expression of p63 increased significantly. In the in vitro serum-free culture condition, the modified explant culture could significantly promote the growth of corneal epithelial stem cells, and expand corneal epithelial stem cells in vitro, which could provide sufficient seed cells for enriching corneal epithelial stem cells and constructing human limbal multilayered epithelial sheets.

3.
Journal of Pharmaceutical Practice ; (6): 52-56, 2020.
Article in Chinese | WPRIM | ID: wpr-782384

ABSTRACT

Objective To designe, synthesize a series of chlorin p6 ether photosensitizers and preliminarily investigate their photodynamic antitumor activity based on previous research results that alkoxyl ether derivatives of 3-vinyl on chlorin f exhibited stronger photosensitive antitumor activity than parent compound. Methods Purpurin-18 (4) was obtained by oxidative degradation with air and alkali on pheophorbide a (5) which was prepared through acid hydrolysis of chlorophyll a from crude chlorophyll extracts in Chinese traditional herb named Silkworm excrement. Then, chlorin p6 trimethylester (2) were formed via basic hydrolysis of internal anhydride ring for lead compound 3 and following immediately methylation with CH2N2. The intermediate 2 reacted with 33% HBr, following nucleophilic substitution with various alkoxyl alcohol to get six title compounds (1). All title compounds were subjected to photodynamic antitumor activity screening for melanoma B16-F10 cell in vitro. Results All title compounds showed much higher phototoxicity against melanoma B16-F10 cells than talaporfin and verteporfin. Their structures were confirmed by 1H-NMR, 13C-NMR, ESI-MS and ESI-HRMS spectra. Conclusion Chlorin p6 ether compounds were promising candidate photosensitizers for PDT applications due to theirs high dark toxicity/phototoxicity ratio and excellent phototoxicity, which were worthy of further research and development.

4.
Chinese Journal of Biotechnology ; (12): 901-909, 2019.
Article in Chinese | WPRIM | ID: wpr-771320

ABSTRACT

To establish a novel colloidal gold immunochromatography assay (GICA) for rapid, sensitive and accurate detection of Haemophilus influenzae infection by using the outer membrane protein P6 as detection target. First, the linear antigen epitope located in the extracellular domain of the P6 protein (GenBank accession number: AGH02799) was predicted by bioinformatics analysis. The region (62-75 aa of the protein) with strong antigen specificity was chosen and synthesized. Two rabbits were then immunized by the polypeptides (14 aa) for production of polyclonal antibodies. Then, the recombinant P6 proteins were also obtained to produce polyclonal antibodies. Finally, based on the two antibodies, a novel colloidal GICA for detection of Haemophilus influenzae infection was established and the specificity, sensitivity, repeatability and stability of this method were evaluated. At the same time, the method was tested in clinical simulation, and the plate culture method was used to verify its accuracy. The test strip for Haemophilus influenzae infection was successfully prepared. The detection limit of the test strip was as low as 1×105 CFU/mL and the whole process can be completed within 15 minutes. The strip specifically recognized Haemophilus influenzae and did not react with nine of other common respiratory pathogens such as Streptococcus pneumoniae, Moraxella catarrhalis, Mycoplasma pneumonia, and Legionella pneumophila. And the strips could be stored at 25 °C for at least 6 months without losing sensitivity or specificity. The coincidence rate between the results of 200 clinical samples and the plate culture method was 90.5%. Haemophilus influenzae protein P6, which possessed a high degree of surface antigen accessibility and antigencity, could be used as a marker for Haemophilus influenzae detection. The immunochromatographic colloidal gold test strip which bears the features of rapidity, convenience and sensitivity provides a unique tool for the on-site surveillance and diagnosis of Haemophilus influenzae infection in clinical test.


Subject(s)
Animals , Humans , Rabbits , Chromatography, Affinity , Diagnostic Tests, Routine , Reference Standards , Gold Colloid , Chemistry , Haemophilus Infections , Diagnosis , Haemophilus influenzae , Limit of Detection , Sensitivity and Specificity
5.
Chinese Journal of Experimental and Clinical Virology ; (6): 195-198, 2018.
Article in Chinese | WPRIM | ID: wpr-806049

ABSTRACT

Objective@#The VP8* core protein of rotavirus P[6] genotype LL4260 was purified by prokaryotic expression, which is important for further study of protein structure and function.@*Methods@#The P[6] genotype LL4260 strain was obtained by PCR.The recombinant plasmid pET30 a-LL4260VP8*core was inserted into pET30 a vector and transformed into BL21 (DE3) competent cells with the correct recombinant plasmid. The expressed protein is purified by affinity chromatography and molecular sieve chromatography.@*Results@#The pET30 a-LL4260VP8* core region protein is soluble in the supernatant and proteins of approximately 22 kDa are identified by electrophoresis using purified proteins.@*Conclusions@#In this study, LL4260 containing pET30 a-LL4260VP8* core plasmid was successfully constructed and LL4260 strain VP8* protein was expressed.

6.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 481-486, 2017.
Article in English | WPRIM | ID: wpr-812090

ABSTRACT

Most of Chinese medicinal herbs are subjected to traditional processing procedures, including stir-frying, charring, steaming, boiling, and calcining before they are released into dispensaries. The marketing and identification of processed medicinal materials is a growing issue in the marketplace. However, conventional methods of identification have limitations, while DNA mini-barcoding, based on the sequencing of a short-standardized region, has received considerable attention as a new potential means to identify processed medicinal materials. In the present study, six DNA barcode loci including ITS2, psbA-trnH, rbcL, matK, trnL (UAA) intron and its P6 loop, were employed for the authentication of 45 processed samples belonging to 15 species. We evaluated the amplification efficiency of each locus. We also examined the identification accuracy of the potential mini-barcode locus, of trnL (UAA) intron P6 loop. Our results showed that the five primary barcode loci were successfully amplified in only 8.89%-20% of the processed samples, while the amplification rates of the trnL (UAA) intron P6 loop were higher, at 75.56% successful amplification. We compared the mini-barcode sequences with Genbank using the Blast program. The analysis showed that 45.23% samples could be identified to genus level, while only one sample could be identified to the species level. We conclude that trnL (UAA) p6 loop is a candidate mini-barcode that has shown its potential and may become a universal mini-barcode as complementary barcode for authenticity testing and will play an important role in medicinal materials control.


Subject(s)
DNA Barcoding, Taxonomic , Methods , DNA, Plant , Genetics , Discriminant Analysis , Drugs, Chinese Herbal , Chemistry , Classification , Introns , Plant Proteins , Genetics , Plants, Medicinal , Chemistry , Classification , Genetics
7.
Chinese Journal of Zoonoses ; (12): 1053-1059, 2017.
Article in Chinese | WPRIM | ID: wpr-703042

ABSTRACT

The aim of this study is to investigate the distribution and sequence conservation of P6 outer membrane protein (OMP6)-encoding gene of nontypeable Haemophilus influenzae isolates as well as screen and identify the predominant T-and B-cell (T-B)-combined antigenic epitopes in OMP6 sequences and their immunogenicity.The entire omp6 genes of NTHi isolates were amplified by PCR and the amplification products were sequenced after T-A cloning.By using bioinformatic softwares,the sequence conservation and membrane location of OMP6 were analyzed as well as the T-B-combined antigenic epitopes in OMP6 were predicted.The immunogenicity and immunoreactivity of T-B-combined antigenic epitope peptides displayed by recombinant phage PⅢ proteins (rPⅢ) were determined by Western Blot assay and ELISA.The PCR showed that all the 35 NTHi isolates tested were detectable for omp6 gene.The identities of nucleotide and amino acid sequences of omp6 genes from 28 strains in the NTHi isolates were 98.3%-100% and 99.3 %-100%,respectively.OMP6 of NTHi was predicted as an outer membrane superficial protein that contains OMP6-2-25,OMP6-61-86 and OMP6-98-126 predicted T-B-combined antigenic epitopes.The immunoblotting assay and ELISA confirmed that OMP6-2-25 presented stronger hybridization band with NTHi antisera while 96.9% (59/62),69.4% (43/62) and 74.2% (46/62) of serum samples from NTHi-infected children were positive for OMP6-2-25,OMP6-61-86 and OMP6-98-126 T-B-combined antigenic epitope peptides,respectively.All the results lead to a conclusion thatomp6 is an extensive distribution and sequence conserved gene of NTHi,and OMP6-2-25 is the predominant T-B-combined antigenic epitopes which can be used as the candidates for developing multiple antigenic peptide vaccine against NTHi.

8.
Journal of Pharmaceutical Practice ; (6): 26-30,35, 2017.
Article in Chinese | WPRIM | ID: wpr-790691

ABSTRACT

Objective To design and prepare 13 ,15-cycloimides chlorin p6 (1) ,a class of chlorin related antitumor photo-sensitizers ,which contain a more stable six-membered cyclic imide comparing to the exocyclic anhydride ring of purpurin-18 (2) .Compounds (1) exhibit strong absorption at long wavelengths near λmax 700 nm to take full advantage of greater tissue penetration .Methods Pheophorbide a (3) was obtained by acid hydrolysis of chlorophyll a ,which was from crude chlorophyll extracts of Chinese traditional herb named Silkworm excrement .Purpurin-18 (2) was prepared by air oxidation and alkali open loop simultaneously on five-membered beta-keto carboxylic ester ring of pheophorbide a (3) .Finally ,the target compounds 1a~1j were synthesized via condensation of its anhydride ring with various amines including carboxyl-protected amino acids . Results Target compounds 1a~1j were successfully synthesized in yields ranged from 32 .6% to 65 .2% .Their structures were confirmed by elemental analysis ,ESI-MS and 1 H NMR spectra .Conclusion Treatment of purpurin-18 (2) with amines can produce target compounds 1a~1j .The starting raw material was inexpensive and readily available .The reaction conditions were mild and workup was convinient .

9.
Chinese Pharmaceutical Journal ; (24): 1485-1489, 2015.
Article in Chinese | WPRIM | ID: wpr-859611

ABSTRACT

OBJECTIVE: To identify the processed products and raw materials of Cistanche tubulosa, Chrysanthemum morifolium and Panax ginseng by DNA barcoding technology based on ITS2 and P6 loop of trnL intron sequences. METHODS: The genomic DNA from all the samples were extracted, amplified and sequenced. The obtained sequences were aligned using MEGA5.0 and Blast method. RESULTS: The ITS2 sequences could identify Cistanche tubulosa and Panax ginseng accurately, while the P6 loop of trnL intron sequences were amplified successfully in highly degraded DNA samples. The P6 loop sequences from the processed products and raw materials of Cistanche tubulosa, Chrysanthemum morifolium and Panax ginseng were identical with those of their herbs, but processed products can not be identified to species level by the similarity search method. CONCLUSION: ITS2 Sequence can be used as the suitable DNA barcodes for identification of the herbs oi Cistanche tubulosa, and Chrysanthemum morifolium, meanwhile, the P6 loop of trnL intron sequence holds a bright prospect in quality inspection of traditional Chinese medicines and foods.

10.
Chinese Pharmaceutical Journal ; (24): 1261-1265, 2015.
Article in Chinese | WPRIM | ID: wpr-859569

ABSTRACT

OBJECTIVE: To choose suitable barcodes for Draconis Sanguis and find the new method to identify Draconis Sanguis. METHODS: The genomic DNAs from 41 samples were extracted. Draconis Sanguis samples were amplified and sequenced based on seven candidate DNA barcodes [ITS, ITS2, psbA-trnH, rbcL, matK, trnh (UAA) intron, P6 loop of trnh (UAA) intron]. The obtained sequences were assembled using the CodonCode Aligner V 4.2. The Kimura 2-Parameter (K2P) distances were calculated using MEGA 5.0. The neighbor-joining (NJ) phylogenetic trees were constructed. RESULTS: Six out of the seven candidates [ITS, ITS2, psbA-trnH, rbcL, matK, trnh (UAA) intron] were not able to be amplified, while P6 loop of trnh (UAA) intron could be used for identification. The length of the P6 loop of trnh( UAA) intron sequence of Daemonorops draco was 45 bp. The maximum K2P intra-spe-cific genetic distances of Daemonorops draco were much smaller than the minimum inter-specific genetic distances between Daemonorops draco and its adulterants. The NJ tree showed that Draconis Sanguis differed from its adulterants obviously. CONCLUSION: The P6 loop of trnL (UAA) intron can be used as DNA barcode for the identification of Draconis Sanguis.

11.
Rev. med. vet. zoot ; 61(2): 164-170, may.-ago. 2014. ilus
Article in English | LILACS-Express | LILACS | ID: lil-726952

ABSTRACT

Bovine cervical mucus (BCM) is important for selection and transport of spermatozoa. When air-dried, BCM obtained at oestrus exhibits arborescent crystallizations, among other arrangements. Considering the relevant endocrine and reproductive information indirectly obtained from BCM crystallization, a morphological investigation was carried out to study its crystalline patterns. BCM samples were collected from healthy Holstein Friesian heifers at oestrus, their crystalline patterns photographed and its morphology analyzed. The majority of the crystallizations obtained showed the typical tree-like patterns reported for BCM. However, a highly symmetrical arrangement was found, characterized by a star-like morphology with six straight, highly defined axes that protrude from the same central point, forming 60º angles. In terms of current knowledge, this short report is the first to show this crystallization geometry in BCM, which, additionally, is remarkably similar to P6B mucus reported for periovulatory human cervical mucus. Even though the role of mucus presenting this type of crystallization is as yet unknown for bovines, its possible functions are also briefly discussed here.


El moco cervical bovino es importante para la selección y el transporte espermático. El moco, obtenido durante el estro y secado al aire, exhibe cristalizaciones con formas principalmente arborescentes. Considerando la importante información endocrina y reproductiva que es posible obtener a partir de la cristalización del moco cervical, se efectuó una investigación morfológica con el propósito de estudiar sus patrones cristalinos. Las muestras de moco se obtuvieron de novillas Holstein Friesian en estro; posteriormente, los patrones de cristalización del moco fueron fotografiados para finalmente analizar su morfología. Las cristalizaciones obtenidas correspondieron a típicos patrones arboriformes previamente reportados. Sin embargo, lo que llamó la atención fue el hallazgo de un arreglo altamente simétrico en una novilla, caracterizado por una morfología similar a estrella con seis ejes rectos, bien definidos, que surgen desde el mismo punto central y forman ángulos de 60º. Según nuestro conocimiento, esta comunicación breve reporta por primera vez la presencia de dicha geometría de cristalización en vaquillas, la cual es muy semejante al patrón cristalino subtipo P6 reportado para el moco cervical perioBvulatorio humano. Si bien el rol ejercido por este tipo de cristalización de moco aún se desconoce en bovinos, se discuten aquí sus posibles funciones.

12.
The Journal of Clinical Anesthesiology ; (12): 592-593, 2014.
Article in Chinese | WPRIM | ID: wpr-452239

ABSTRACT

Objective To evaluate the effect of the electronic anti-nausea instrument on the postoperative nausea and vomiting of patients with gynecological laparoscopic surgery.Methods One hundred and eighty patients for gynecological laparoscopic surgery were enrolled and randomized into 2 groups with 90 patients in each.Patients in group T accepted patient-control transcutaneous elec-troacupoint stimulation at P6 (Neiguan)point from the time before the induction of anesthesia to 24 h after surgery.Patients in group C accepted the same device of electronic anti-nausea instrument with-out transcutaneous acupoint stimulation.Data were recorded of the nausea and vomiting in postopera-tive 2,6,12 and 24 h respectively.Results The incidence and severity of nausea at 6,12 and 24 h and vomiting at 6,24 h after operation in group T were both lower than those in group C(P < 0.05 ). Conclusion With patient-control transcutaneous acupoint stimulation at P6 point,the incidence of both early PONV and late PONV are reduced in patients with gynecological laparoscopic surgery.

13.
Chinese Journal of Microbiology and Immunology ; (12): 624-629, 2014.
Article in Chinese | WPRIM | ID: wpr-456259

ABSTRACT

Objective To measure the concentrations of antibodies against outer membrane protein P6 and its T-and B-combined antigenic epitopes of nontypeable Haemophilus influenzae ( NTHi) in children and adults of different ages and to evaluate the differences among different subjects for further investigation on NTHi multiple antigenic peptide vaccine .Methods A prokaryotic expression system was established to ex-press the recombinant outer membrane protein P 6 of NTHi.The expressed protein was purified by using Ni-NTA affinity chromatography .T-and B-cell epitopes in protein P6 were predicted with Epitope prediction software 1.0 and ANTIGENIC program and were used to synthesize T-and B-combined antigenic epitopes .A total of 605 subjects aged from 1 day to 103 years old were recruited from October 2013 to March 2014 .Ser-um concentrations of antibodies against protein P 6 and its T-and B-combined antigenic epitopes were meas-ured by using ELISA .Mann-Whitney U test was used to analyze the differences between groups .Pearson product-moment correlation coefficient was used for correlation analysis .Results Four T-and B-combined antigenic epitopes including P 6-2, P6-61, P6-95 and P6-122 were predicted and synthesized .The levels of antibodies against NTHi P6 and P6-2, P6-61, P6-95 and P6-122 were significant lower in the <1 months group than those in the 1-6 months group (all P<0.001) and 7 months-3 years group (all P<0.001).Three groups including 7 months-3 years group , 4-6 years group and 7-14 years group showed significant differ-ences regarding to the antibodies levels , among the 7 months-3 years group showed the highest levels , fol-lowed by the 4-6 years group and the 7-14 years group.However, no significant difference was found be-tween other adjacent groups .Concentrations of antibodies against P 6-2, P6-61, P6-95 and P6-122 were pos-itively correlated with the level of antibody against P 6 (P<0.0001).Conclusion The distribution of anti-bodies against T-and B-combined antigenic epitopes in P6 was highly in accord with those against P6, which indicated good immunogenicity of those epitopes .The highest antibodies levels were found in subjects aged 7 months to 3 years old , which might correlate with the high risk of NTHi infection at that stage .

14.
Virologica Sinica ; (6): 57-68, 2012.
Article in Chinese | WPRIM | ID: wpr-423947

ABSTRACT

Protamines are a group of highly basic proteins first discovered in spermatozoon that allow for denser packaging of DNA than histones and will result in down-regulation of gene transcription[1].It is well recognized that the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) encodes P6.9,a protamine-like protein that forms the viral subnucleosome through binding to the viral genome[29].Previous research demonstrates that P6.9 is essential for viral nucleocapsid assembly,while it has no influence on viral genome replication[31].In the present study,the role of P6.9 in viral gene transcription regulation is characterized.In contrast to protamines or other protamine-like proteins that usually down-regulate gene transcription,P6.9 appears to up-regulate viral gene transcription at 12-24 hours post infection (hpi),whereas it is non-essential for the basal level of viral gene transcription.Fluorescence microscopy reveals the P6.9's co-localization with DNA is temporally and spatially synchronized with P6.9's impact on viral gene transcription,indicating the P6.9-DNA association contributes to transcription regulation.Chromatin fractionation assay further reveals an unexpected co-existence of P6.9 and host RNA polymerase Ⅱ in the same transcriptionally active chromatin fraction at 24 hpi,which may probably contribute to viral gene transcription up-regulation in the late infection phase.

15.
Chinese Journal of Microbiology and Immunology ; (12): 371-375, 2011.
Article in Chinese | WPRIM | ID: wpr-415641

ABSTRACT

Objective To detect the difference of cytokines and antibodies productions by immunologic system from mice vaccinated with recombinant P6 protein with different immunization routes and adiuvants.Methods 6 weeks female BALB/c mice were vaccinated with meombinant protein P6 combined with A1(OH)3,CpG ODN or the mix of them through intramuscular and intranasal.The mice were boosted twice with the same dose by the same route.Serum and respiratory tract specimen were collected to detect rP6 specific antibodies by ELISA.Results In the 6th week after immunization.The higher titer of serum rP6 specific IgG antibody were detected from the two groups vaccinated with A1(OH)3+CpG ODN+rP6 by intramuscular and vaccinated with CpG ODN+rP6 by intranasal(F=41.259.P=0.000).The ELISPOT experiment showed that,Inoculation with the CpG ODN+rP6 either by intranasal or intramuscular immunization could induce a large number of antigen-specific T lymphocyte activation.In addition.The mice vaccinated with the CpG ODN+rP6 through intranasal can be detected high titer rP6 specific mucosal IgA antibody(F=70.966.P=0.000).Conclusion Inoculation with the CoG ODN+rP6 by jntranasal immunization not only can induce high-titer serum IgG antibody,but also can induce high-titer mucosal IsA antibody and CD4+Th1.CD8+T lymphoeyte activation.

16.
Chinese Journal of Microbiology and Immunology ; (12): 349-354, 2010.
Article in Chinese | WPRIM | ID: wpr-379861

ABSTRACT

Objective To clone and express recombinant outer membrane protein P6, determine its optimal expression conditions and to investigate the immunoprotective effects of the P6 protein on mice. Methods The P6 gene of nontypeable Haemophilus influertzae(NTHi) was amplified by PCR from the NTHi genome and cloned into expression vector pET-32a (+) to generate the pET-32a-P6 recombinants. They were confirmed by nuclease digestion and sequence analysis. The verified recombinant was transformed into E. coli BL21 (DE3). Its optimal expression conditions were determined such as engineering strains, the concentration of IPTG, inducing temperature, inducing time, different medium etc. The recombinant protein was purified by Q Sepharose~(TM) XL ion exchange and gel filtration chromatography. The protein was analyzed by SDS-PAGE, Western blot and sequencing. BALB/c mice were immunized with recombinant protein be-fore challenged by NTHi through intraperitoneal injection. Then the mortality rate of different group was com-pared. Results The recombinant P6 of NTHi was successfully constructed and expressed in E. coli at a rel-atively high level. The purity was up to 95% after purification. The relative molecular mass of the protein is 14 145. 848. The recombinant protein was confirmed to show specific reaction on the antiserum through Western blot. The animal experiments showed the mortality rates of immunization groups were significantly lower than that of the control group (P < 0.05). Conclusion The successful expression of the recombinant P6 will be very helpful for the further study on development of vaccine, its purified, immunological activity and antibody preparation.

17.
Korean Journal of Anesthesiology ; : 413-418, 2009.
Article in Korean | WPRIM | ID: wpr-179768

ABSTRACT

BACKGROUND: Postoperative nausea and vomiting (PONV) is a common problem in patients recovering from anesthesia and surgery. P6 point is the acupressure point for prevention of postoperative nausea and vomiting. We evaluated the efficacy of acupressure at the P6 point in 94 patients undergoing thyroidectomy in a randomized, prospective and placebo-controlled study. METHODS: Ninety-four female patients, aged 18 to 60, scheduled for elective thyroidectomy, were randomized to have either placebo band or acupressure band (Sea-Band(R) UK Ltd., Leicestershire, England, UK) applied to the P6 point of both hands before induction of anesthesia. The acupressure bands removed 24 h later. Postoperative nausea and vomiting was evaluated 1, 6 and 24 h following surgery. In addition, the need for rescue antiemetic medication during 24 h was registered. RESULTS: The incidence of postoperative nausea was lower in acupressure group at 0-1 h (16.7% vs. 39.1%; P = 0.015) and at 6-24 h (0% vs. 15.2%; P = 0.05). The need for rescue antiemetic medication was also lower at 0-1 h (4.2% vs. 23.9%; P = 0.006), at 1-6 h (6.2% vs. 20.9%; P = 0.039) and at 6-24 h (0% vs. 13%; P = 0.012). CONCLUSIONS: In patients undergoing thyroidectomy, nausea and need of rescue antiemetic medication were reduced by acupressure at the P6 point.


Subject(s)
Aged , Female , Humans , Acupressure , Anesthesia , England , Hand , Incidence , Nausea , Postoperative Nausea and Vomiting , Prospective Studies , Thyroidectomy
18.
Journal of Preventive Medicine ; : 72-79, 2008.
Article in Vietnamese | WPRIM | ID: wpr-915

ABSTRACT

Background: Currently, the World Health Organization is encouraging developing countries to establish a seed lot system of rotavirus vaccine for production of this vaccine. Objectives: To determine gene sequences of rotavirus strain that was used for vaccine production and to evaluate its stability. Materials and method: Master seed (G4P6MS), Working seed (G4P6WS) and vaccine strain (G4P6VX) of Rotavirus were used for analysis at the US Center for Disease Control and Prevention (CDC). Results: 855 base pairs of gene 4 (VP4); 1195 base pairs of gene 6 (VP6); 824 base pair of gene 9 (VP7) and 715 base pairs of gene 10 (NSP4) from seed lot system and vaccines of G4P6 strain were determined. The results demonstrated this seed lot system is completely stable during vaccine production. There is no difference for nucleotide and amino acid sequence in this seed lot system. Conclusion: G4P6 strain (2001019203) is completely stable during vaccine production.


Subject(s)
Rotavirus Vaccines
19.
The Korean Journal of Laboratory Medicine ; : 86-92, 2006.
Article in Korean | WPRIM | ID: wpr-216295

ABSTRACT

BACKGROUND: Group A rotavirus is a major cause of severe diarrhea in children throughout the world. For the proper management of rotavirus infections, it will be helpful to know their clinical characteristics according to the G and P genotypes of the infecting virus. METHODS: The diarrheal stool specimens from patients hospitalized in Chosun University Hospital during 2002-2003 were tested for rotavirus by Dipstick 'Eiken' Rota kit. Rotavirus antigen-positive stool specimens were analyzed for group A rotavirus by RT-PCR, and the group A-positive PCR products were genotyped for P and G types by PCR. RESULTS: Among the 119 specimens analyzed for genotypes, the predominant strain was genotype G4P[6] (51.3%), followed by G2P[4] (19.3%), G1P[8] (7.6%), G3P[8] (5.0%), and G9P[8] (4.2%). To examine the characteristics of each rotavirus genotype, a clinico-epidemiological study was performed for 100 patients whose medical records were available. The frequencies of diarrhea, vomiting, dehydration, and fever; the rates of nosocomial infection and transfer from other hospitals; and the mean severity scores were significantly different among the patients infected with different types of rotavirus. Especially, patients with G4P[6] type were more likely than those infected with other genotypes to show the following distinct features: Most patients showed milder symptoms and were neonates transferred from other obstetric hospitals and 68.4% of the cases were nosocomial infection. G4P[6] strains were isolated almost all along the year. The mean severity scores of patients infected by G4P[6], G2P[4], G1P[8], G3P[8], and G9P[8] strains were 6.8, 9.5, 8.0, 9.0, and 10.8, respectively. CONCLUSIONS: Many features of rotavirus infections including the epidemic period, rate of nosocomial infection, age and severity of symptoms were different according to the genotypes of the infecting virus.


Subject(s)
Child , Humans , Infant, Newborn , Cross Infection , Dehydration , Diarrhea , Fever , Gastroenteritis , Genotype , Medical Records , Polymerase Chain Reaction , Rotavirus Infections , Rotavirus , Vomiting
20.
Korean Journal of Anesthesiology ; : 743-748, 2003.
Article in Korean | WPRIM | ID: wpr-82797

ABSTRACT

BACKGROUND: Postoperative nausea and vomiting is one of the main problems after mastoidectomy and tympanoplasty. There is a growing interest in the use of nonpharmacologic and nonintravenous approaches to the prevention of postoperative nausea and vomiting. The aim of this study was to investigate the effect of stimulating the P6 acupoint and of the use of metoclopramide intranasal spray on the prevention of postoperative nausea and vomiting after mastoidectomy and tympanoplasty. METHODS: We studied 60 patients who received mastoidectomy and tympanoplasty for chronic ottitis media. No antiemetic agent or device was provided in the C group (n = 20). Acupressure on the P6 acupoint was applied after surgery in the P6 group (n = 20). In the M group (n = 20), metoclopramide was sprayed intranasally before extubation. Severity values of postoperative nausea and vomiting were assessed using 5 scales at different postoperative times. RESULTS: The severity of postoperative nausea and vomiting was significantly lower in the P6 group than in the C and M groups. There was no difference in the severity of postoperative nausea and vomiting between the C and M groups. At a postoperative 8 and 16 hr, there was a statiscally significant decrease of the severity of postoperative nausea and vomiting in the P6 group. CONCLUSIONS: Acupressure on the P6 acupoint reduced the incidence and severity of postoperative nausea and vomiting after mastoidectiomy and tympanoplasty. This result suggests that acupressure at P6 may be a useful new nonpharmacologic approach to the reduction or prevention of postoperative nausea and vomiting after mastoidectomy and tympanoplasty.


Subject(s)
Humans , Acupressure , Acupuncture Points , Incidence , Metoclopramide , Nausea , Postoperative Nausea and Vomiting , Tympanoplasty , Vomiting , Weights and Measures
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