Polymerase chain reaction (PCR) is one of the common techniques in molecular biology, which can amplify nucleic acids through the cycle of denaturation, annealing and extension. Based on the principle of common PCR, rapid PCR is to realize the amplification of nucleic acids in less time without affecting the specificity, sensitivity and fidelity of the reaction. A lot of research work in this field has been going on in recent years. This article will make a review of the development of rapid PCR with emphases on the improvement of DNA polymerase, the choice of additives and the improvement of thermocyclers.

Objective To analyze the mutations in nucleotide sequences of transfusion transmitted virus(TTV) in neonatal infection.Methods Neonatal serum TTV-DNA was detected by a nested PCR technique.Fifteen Chinese neonates with positive TTV-DNA were diagnosed as TTV infection.ORF1 sequences of TTV-DNA from these neonates were determined.Results Homology of Chinese TTV(C01-C15) and Japanese TTV(N22)isolated ranged from 87.1%-97.7% at nucleotide level,but there were point mutations in Chinese TTV,such as GG→TT in locus 112 and 113,TTATC→CCTAT in locus 236-240.Conclusions Chinese and Japanese TTV isolated had the same genotype.Some gene mutations may increase the TTV pathogen,and result in neonatal hepatitis syndrome or hyperbilirubinemia.

Antigens, CD20 ; metabolism ; Carcinoma in Situ ; classification ; diagnosis ; metabolism ; pathology ; Diagnosis, Differential ; Humans ; Hyaluronan Receptors ; metabolism ; Hyperplasia ; Precancerous Conditions ; diagnosis ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism ; Urinary Bladder ; metabolism ; pathology ; Urinary Bladder Neoplasms ; classification ; diagnosis ; metabolism ; pathology ; Urothelium ; metabolism ; pathology

Diagnosis, Differential ; Hemangiopericytoma ; metabolism ; pathology ; Hemangiosarcoma ; metabolism ; pathology ; Histiocytoma, Malignant Fibrous ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Leiomyosarcoma ; metabolism ; pathology ; Nerve Sheath Neoplasms ; metabolism ; pathology ; Neuroectodermal Tumors, Primitive ; metabolism ; pathology ; Rhabdoid Tumor ; metabolism ; pathology ; Rhabdomyosarcoma ; metabolism ; pathology ; Soft Tissue Neoplasms ; metabolism ; pathology ; Urinary Bladder ; metabolism ; pathology

Adult ; Dendritic Cell Sarcoma, Interdigitating ; complications ; surgery ; Humans ; Male ; Paraneoplastic Syndromes ; etiology ; Pemphigus ; etiology

Aged ; B-Lymphocytes ; pathology ; Histiocytic Sarcoma ; complications ; Humans ; Lymphoma, B-Cell ; etiology ; Lymphoma, Large B-Cell, Diffuse ; etiology ; pathology ; Male

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.
Animals ; Half-Life ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Molecular Probes ; pharmacokinetics ; RNA, Small Interfering ; chemistry ; Rabbits

AIM:To investigate the effects of 1.8mm coaxial micro incision phacoemulsification on corneal endothelial injury and postoperative visual acuity.METHODS: Totally 145 eyes in 120 patients underwent phacoemulsification from July 2013 to July 2015 were randomly divided into observation group 60 cases (73 eyes) and control group 60 cases (72 eyes).The observation group 60 cases were given 1.8mm coaxial micro incision cataract phacoemulsification operation,while the control group were given traditional 3.2mm coaxial micro incision cataract surgery.The uncorrected visual acuity (UCVA),best corrected visual acuity (BCVA),corneal thickness of incision area,incision width,incision length,macular retinal thickness,surgically induced astigmatism,corneal endothelial cell counts and complications of the two groups were compared.RESULTS: The UCVA and BCVA on 1wk after surgery of the observation group were significantly higher than the control group (t=3.604,7.109;P<0.05);the width of incision on 1wk and 1mo after surgery of the observation group were significantly less than the control group (t=205.3,225.2;P<0.05).The length of incision in observation group was significantly greater than the control group (t=3.926,5.009;P<0.05).Macular retinal thickness 1wk after surgery of the observation group was significantly less than the control group (t=2.817,P<0.05).The surgically induced astigmatism was significantly less than the control group (t=19.43,22.16;P<0.01);the difference of corneal edema between the two groups was not significant (8.22% vs 11.11%) (x2=0.348,P>0.05).CONCLUSION: The 1.8mm micro incision phacoemulsification is helpful to improve the visual acuity of patients with cataract phacoemulsification,which may be related to the reduction of corneal cell injury,enhancement of corneal closure and decrease post-operation corneal original astigmatism.

OBJECTIVETo evaluate the clinical efficacy of treating myelodysplastic syndrome (MDS) by hematopoietic stem cell transplantation (HSCT) combined with Chinese medical syndrome typing.

METHODSFrom July 2009 to July 2013, 6 MDS patients were treated with allo-HSCT combined with Chinese medical syndrome typing from HLA-identical sibling donors at Department of Hematology, Zhejiang Provincial Hospital of Chinese Medicine. Patients were classified as refractory anemia (RA, 2 cases), refractory anemia with ringed sideroblast (RARS, 1 case), refractory cytopenia with multilineage dysplasia (RCMD, 2 cases), and RA with excess blasts-I (RAEB-I , 1 case). Modified BuCy conditioning regimen was used in all 6 cases. Two patients received bone marrow transplantation (BMT), 1 patient received peripheral blood stem cell transplantation (PBSCT), and 3 patients received BMT + PBSCT. In order to prevent the occurrence of graft-versus-host disease (GVHD), all patients were treated with cyclosporine + methotrexate + mycophenolate mofetil. Different Chinese medical treatment methods (by syndrome typing) were given to patients according to different criticality of international prognostic scoring system (IPSS, 5 at moderate risk and 1 at high risk).

RESULTSAll 6 patients successfully reconstructed their hematopoietic system. The time from transplantation to ANC ≥ 0.5 x 10(9)/L and platelet (PLT) ≥ 20 x10(9)/L were 13 (9-15) days and 11 (9-22) days respectively. Main complications were GVHD. Acute GVHD (aGVHD) occurred in 4 cases, 3 cases of grade I and 1 case of grade II, and local chronic GVHD (cGVHD) occurred in 1 patient. All cases survived with median follow-up of 18 (11-58) months. The overall survival (OS) and disease-free survival (DFS) rate were 100%.

CONCLUSIONSHSCT combined with Chinese medical syndrome typing could improve clinical symptoms, reduce transplant as- sociated complications. So it was an effective treatment choice for MDS.

Biomedical Research ; Blood Platelets ; Bone Marrow Transplantation ; Cyclosporine ; therapeutic use ; Disease-Free Survival ; Drugs, Chinese Herbal ; therapeutic use ; Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Humans ; Medicine, Chinese Traditional ; Methotrexate ; therapeutic use ; Myelodysplastic Syndromes ; therapy ; Transplantation Conditioning ; Transplantation, Homologous ; Treatment Outcome

Traditional Chinese medicine (TCM) dispensing is the final step of TCM used for clinical treatment, the stability of TCM dispensing is the guarantee of good clinical effect. Establishment of effect-constituent equivalence for Chinese herbal pieces based on clinical efficacy, can not only guarantee the stability of TCM dispensing, but also relate to the precision of clinical effect. This study chose Coptidis Rhizoma as the model, established effect-constituent equivalence of Coptidis Rhizoma, based on the effect-constituent index already established by our research group, and taking into consideration of homogeneity of clinical dosage and compliance of decoction, the uniformity of dispensing for different specification of Coptidis Rhizoma decoction pieces was studied. This research model was then applied to guide the specification-optimization of Coptidis Rhizoma and its clinical dispensing. The result indicated, effective constituent equivalence could reflect the fluctuation of specification, dosage and decoction to the fluctuation of efficacy; Optimized Coptidis Rhizoma decoction pieces had the characteristic of high homogeneity as for clinical dispensing, good compliance as for decoction, and high effective constituent equivalence. In conclusion, effective constituent equivalence could improve relevance of methods of TCM dispensing control to clinical effect. Preparated Superior-standard Decoction Pieces based on effective constituent equivalence was featured by good quality and a good practice of adjustable dosage, which could promote the development of TCM decoction pieces toward precision medicine.
Chemistry, Pharmaceutical ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Rhizome ; chemistry ; Therapeutic Equivalency