Objective: This study aimed to observe the long-term effect of Pseudomonas aeruginosa preparation used in peritoneal injection of advanced colorectal cancer patients during surgery. Methods: A total of 83 colorectal cancer patients who received surgery between September 2006 and March 2008 were enrolled in this study. The patients were divided into two groups. Palliative resection and a 10 ml P. aeruginosa peritoneal injection were performed in 30 of 83 patients in the treatment group. Simple palliative resection was conducted in the other 53 patients, which comprised the control group. Both groups were then treated by regular chemotherapy and radiotherapy. Results:The follow-up visit was completed in 79 of 83 patients, with a high follow-up rate of 95.2%. No significant difference was found in the five-year overall survival time between the two groups (P=0.403). However, the five-year median survival time in the control group was only 13.9 ± 2.14 months, whereas that in the treatment group was 17.2 ± 2.12 months. Conclusion: Within a short period, peritoneal injection of P. aeruginosa during surgery could confer certain survival advantages for advanced colorectal cancer patients. However, the long-term effect of this therapy remains unknown.

Objective To investigate the gene therapy of hypoparathyroidism (HPT) with hemopoietic stem cell s(HSCs) integrated with parathyroid hormone (PTH) gene.Methods Human PTH gene was amplified by polymerase chain reaction from pcDNA 3.1 -PTH templates and inserted into murine stem cell virus (MSCV) vectors. The plasmids recombinated with PTH gene were transfected into PA317 packaging cells and screened for positive clones with Geneticin. The condensed recombinant retroviruses were obtained and used to infect HSCs coming from umbilical cord blood. The infected HSCs were injected into the mice suffering from HPT. The change of symptoms and serum level of PTH and calcium in each group of mice was observed.Results Human PTH gene was proved to be inserted into MSCV vectors and the titres were up to 2?10 7 colony forming unit (CFU)/ml in condensed retroviral solution. The secretion of PTH reached 15 ng per 10 6 cells per 48 h. The wild type viruses were not detected by PCR amplification. The mice suffering from HPT recovered quickly and the serum level of calcium and PTH remained normal for about 3 months after the HSCs recombinated with PTH gene were injected into them. In the groups of mice subject to the injection of the recombinant retroviruses containing PTH gene, the serum level of PTH and Calcium was significantly increased in a short time, and then declined slowly. And the mice took on the manifestation of hypoparathyroidism again. The mice only subject to the implantation of HSCs were all dead within 20 days.Conclusion A long term therapeutic effect could be obtained by infusing the HSCs which were integrated with PTH gene into the mice suffered from hypoparathyroidism.

OBJECTIVETo identify the Cynomorii Herba and its analogues species using DNA barcoding technique.

METHODTotal genomic DNA extracted from all materials using the DNA extraction kit. The internal transcribed spacer 2 (ITS2) regions were amplified using polymerase chain reaction (PCR), and purified PCR products were sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner 3.7.1. The Kimura 2-Parameter (K2P) distances and GC content were computed using MEGA 5. 0. Species identification analyses were conducted through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) trees.

RESULTThe ITS2 sequence lengths of Cynomorii Herba were 229 bp. The average intra-specific genetic distances of Cynomorii Herba were 0.003. The average inter-specific genetic distances between Cynomorii Herba and its adulterants species were 0.760. The results showed that the minimum inter-specific divergence is larger than the maximum intra-specific divergence. The species identification system for traditional Chinese medicine and NJ trees results indicated that Cynomorii Herba and its adulterants species can be easily identification.

CONCLUSIONThe ITS2 region is an efficient barcode for identification of Cynomorii Herba, which provide a new technique to ensure clinical safety in utilization of traditional Chinese medicine.

Cynomorium ; classification ; genetics ; DNA Barcoding, Taxonomic ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Polymerase Chain Reaction

OBJECTIVETo study the effect of xenotransplantation with pig parathyroid cells, which was prepared using cell microencapsulation technique, on the treatment of hypoparathyroidism in rats without immunosuppressor.

METHODSParathyroid cells were isolated from 10 healthy newborn pigs and encapsulated in alginate-polylysine-alginate (APA) membranes. Thirty-two aparathyroid Wistar rats were randomly allocated to microcapsule, non-microcapsule, empty microcapsule, and control groups. Each rat was injected intraperitoneally with encapsulated porcine parathyroid cells, free porcine parathyroid cells, empty capsules or 0.9% NaCl, respectively. Total serum calcium and parathyroid hormone levels were monitored continuously for 40 weeks. And then, the transplant beds were retrieved and subjected to morphologic and electron microscopic examination.

RESULTSIn those animals xenotransplanted with microencapsulated porcine parathyroid cells, the calcium and PTH levels were consistently within the normal range during the 40 weeks. In contrast, no therapeutic effects were observed in rats in the non-microcapsule group. Furthermore, neither empty capsules nor 0.9% NaCl were shown to have any effect on the recipient's serum calcium or PTH levels. After 40 weeks, electron microscopic examination demonstrated that the parathyroid cells within the microcapsules had survived well in vivo.

CONCLUSIONSXenotransplantation of microencapsulated newborn pig parathyroid cells can successfully treat hypoparathyroidism in rats without using immunosuppressive drugs. The results of this study show the possible clinical use of microencapsulated porcine parathyroid cells.

Animals ; Animals, Newborn ; Calcium ; blood ; Capsules ; Hypothyroidism ; therapy ; Parathyroid Glands ; cytology ; transplantation ; Parathyroid Hormone ; blood ; Random Allocation ; Rats ; Rats, Wistar ; Swine ; Transplantation, Heterologous ; methods

Fifty-eight samples belonging to 7 species of Arisaematis Rhizoma and its adulterants were collected. The ITS2 locus was employed as a DNA barcode and amplified, sequenced and assembled for all of the collected samples. Then, ITS2 sequences have been annotated using HMM-based method. The intra- and inter-specific variations were calculated and NJ tree was constructed using MEGA 6.0 software. The results showed that inter-specific K2P distances were significantly larger than intra-specific distances for all of the three origin species of Arisaematis Rhizoma. Furthermore, three origin species, Arisaema amurense, A. erubescens and A. heterophyllum, can be respectively formed to be a single branch with high bootstrap values. It is concluded that ITS2 can be used to correctly identify Arisaematis Rhizoma from its adulterants and the application of ITS2 in the identification of traditional Chinese medicine has an important prospective.
Arisaema ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics

Adult ; Chordoma ; pathology ; Female ; Humans ; Nose Neoplasms ; pathology

OBJECTIVETo analyze the fungal composition in Massa Medicata Fermentata based on culture dependent method and independent PCR-SSCP technique.

METHODFungi were directly isolated from Massa Medicata Fermentata samples. The obtained strains were identified according to morphology and DNA sequence. Meanwhile the total fungal DNA was extracted from Massa Medicata Fermentata samples, the cultural independent PCR-SSCP technique based on β-tubulin gene were used to identify the mycobiota.

RESULTAccording to cultural method, Aspergillus flavus and Rhizopus oryzae were present in Massa Medicata Fermentata samples, while A. flavus and A. niger were present in fried Massa Medicata Fermentata samples. In contrast, 5 species were obtained by PCR-SSCP technique, A. flavus was overlapped with fungal taxa derived from culture dependent method; A. ambiguu and A. s ivoriensis were dominant with relative abundance of 57% and 35% respectively, while the relative abundance of A. flavus was as low as 4%. None species was obtained from fried Massa Medicata Fermentata samples.

CONCLUSIONPCR-SSCP based on β-tubulin gene could distinguish fungi into species, culture dependent method combined with culture independent method could better understand the fungal composition associated with Massa Medicata Fermentata fermentation.

Fermentation ; Fungi ; isolation & purification ; Medicine, Chinese Traditional ; Polymerase Chain Reaction ; methods ; Polymorphism, Single-Stranded Conformational ; Tubulin ; genetics

Objective To explore the effect of chromosomal abnormality and polygenic inheritance factor in female children with short stature.Methods 1.Chromosome analysis:peripheral blood was drawn for 1 mL and cultured 72 h to analyze chromosome karyotype (Giemsa Banding ) of peripheral lymphocytes.2.Polygenic factor analysis:the children′s final height were estimated based on their parents average height,and analyzed the distribution characteristics of children′s final height and compared the estimate final height with the actual height.Results Eighty-three cases out of the 364 female children with short stature were chromosomal abnormality(22.80%).Among the 83 cases,the 45,XO and 46,X,i(Xq) occupied 70%.The distribution of children target height shifted left,and the target height of 76 cases was lower than 2 standard deviation (-2 s)and the consistency of target height and actual height reached 20.88%.The target height of 7 cases was lower than 2 standard deviation in those whose chromosome turned out to be abnormal,and the consistency of target height and actual height was 8.43%.Conclusions Chromosomal abnormality is one of the most important etiologic agents causing short stature in female children, and polygenic inheritance is another important etiologic agent.

Objective To study the lysozyme activity in Oncomelania hupensis and observe its inhibitory effect on bacterial growth.Methods Soft tissues of Oncomelania hupensis were initially homogenized and immersed in Tris-HCl-TritonX-114 buffer solution for 24 hours then the supernatant was collected after centrifugation at 10 000 × g for 10 minutes.The supernatant was incubated in a 37 ℃ water bath for 15 minutes and centrifuged again at 2000 × g for 10 minutes.The precipitate was put into ultrafiltration tube (relative retention molecular mass =3000) and centrifuged at 4 ℃,5 000 × g for 30 minute to obtain concentrated enzyme.The protein content,lysozyme activity and the antibacterial effect on Micrococcus lysodeikticus,Shigella dysenteriae,Staphylococcus aureus,Escherichia coli and Candida albicans were measured with bicinchonininc acid(BCA) method,turbidimetric method and agar diffusion (K-B) method,respectively.Results The antibacterial protein lysozyme was identified in gastropod protein concentration of the concentrated enzyme was 3.428 g/L.Average activity,total activity,and specific activity were (760 ± 120) × 103 U/L,(1520 ± 240) × 103 U/L and (221.70 ± 35.00)U/mg,respectively.The enzyme had produced exclusive inhibitory effects on growth of Micrococcus lysodeikticus and Shigella dysenteriae.Average inhibitory diameters were 10-12 and 12-15 mm,respectively.No inhibition zone was observed in saline control,Staphylococcus aureus,Escherichia coli and Candida albicans.Conclusions Lysozyme can be extracted from soft tissues of Oncomelania hupensis with Tris-HCl-TritonX-114 buffer solution,and the enzyme has inhibitory effect on growth of Micrococcus lysodeikticus and Shigella dysenteriae but has no antibacterial effect on Staphylococcus aureus,Escherichia coli and Candida albicans.

Objective To study the structure of class 1 integrons in 90 strains of Pseudomonas aeruginosa isolated during two periods of 1992-1996 and 2003-2005,and to get information about the structure changing of class 1 integrons by comparing their structures in two different periods.Methods Routine PCR and long PCR were performed to amplify the class 1 integrons and the gene cassettes they carried, followed with sequencing and blast via GenBank.Results Thirteen out of 41 strians ioslated during the period of 1992-1996 were positive on class 1 intergrons.Long PCR showed that the class 1 integron was 1868 bp in length and contained 2 resistance genes averagely.Six types of resistance genes of qacEA1 (n=6), sull (n=14),aadA1 (n=2),aadB (n=1),PSE-1 (n=2) and tetA (n=1) were found in these integrons, which consisted of 5 patterns of resistance cassette arrangements.Nineteen strains were proved to carry class 1 integrons in 49 isolates from 2003-2005.The mean DNA sequence length of them was 3383 bp with 3.6 resistant genes in averagely,10 types of resistance genes,qacEA1 (n=18),sull (n=25),aadA1 (n=6), aadB (n=7),aacA4 (n=2),PSE-1 (n=3),VEB-1 (n=4),OXA10 (n=1),cm1 A (n=1) and tetA (n =2),were identified in these integrons,which were composed of 9 patterns of resistance cassette arrangements.Conclusion In terms of produce length and resistance cassettes carried in the integrons, greater complexity is found in the structure of class 1 integrons in strains isolated during 2003-2005 than those during 1992-1996.