Through the experiences in the clinical application of Liu Kai-yun's five-meridian theory of Tuina for children and consulting the relevant literatures, the following academic features of Tuina for children in western Hunan district are summarized in this paper: selecting the acupoints according to the pattern/syndrome differentiation and giving the treatment according to the meridian tropism theory; emphasizing the property of the body constitution and applying the reinforcing and reducing techniques; exquisitely combining the acupoints based on the five-meridian theory; coordinating the opening and closing techniques and regulating yin and yang. All of those provided the reference for the theoretic application of this Tuina school.
Acupuncture Points ; China ; Female ; Fever ; therapy ; Humans ; Infant ; Massage ; education ; Meridians

AIM: To screen and identify the differentially expressed genes in lymphocytes of patients with unstable angina in order to find the molecular mechanism of unstable angina. METHODS: Suppression subtractive hybridizations (SSH) and dot blot hybridizations were performed to screen the relatively differentially expressed genes in lymphocyte RNA between the patients with unstable angina pectoris and stable angina pectoris. The obtained expressed sequence tags (ESTs) were used as probes to perform Reverse Northern blot with forward and reverse suppression products. And the positive ESTs were performed RNA slot hybridization with unstable and stable angina group. The obtained ESTs were sequenced and analyzed using BLAST (nr) at NCBI. RESULTS: Three up-regulated ESTs in the unstable angina group, and one down-regulated EST in the stable angina group were obtained. All of them are sequences of known genes. CONCLUSION: All these ESTs may be associated with the unstablization of plaque of coronary artery in patients with unstable angina.

Differences of some points, levels and angles of the healthy and affected sides of patients with peripheral facial paralysis were picked out according to photographs. Through analysis of the index between the healthy and affected side of the patients and the difference between healthy people and patients, it is approved that those special points, levels and angles, which are called as deviation index of eye and mouth, can evaluate peripheral facial paralysis objectively and judge the degree of deviation. Therefore, it provides references for the diagnosis of facial paralysis and its degree judgement.
Adolescent ; Adult ; Aged ; Child ; Eye ; anatomy & histology ; Facial Paralysis ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Mouth ; anatomy & histology

Objective :To construct and identify a ScFv phage display library against human umbilical cord mesenchymal stem cells.Methods: BALB/c mice were immunized with cultured UC-MSCs.After the third immunization,the total RNA was extracted from the spleen cells of the immunized BALB/c mice and purified by affinity chromatography with mRNA Purification Kit.The heavy-chain and light-chain variable region genes(VH and VL) were amplified by PCR using relevant primers.PCR products of VH and VL genes were cloned into the phagemid vector pSEX81 and electroporated into the XL1-Blue strain of E.coli.The ScFv phage display library against human umbilical cord mesenchymal stem cells was constructed and the capacity of library was measured.The library was panned by three cycles and screened with purified UC-MSCs.The percentage of clones containing a full-length scFv-encoding insert and their diversity was determined for unselected and selected libraries.Results: The amplified fragments of VH and VL genes by RT-PCR were about 399bp and 357bp,respectively.VH and VL genes were all successfully cloned into the phagemid vector pSEX81,which were confirmed by the amplication of 786bp full-length scFv fragments by PCR.The ScFv phage display library had a capacity of approximately 2?107 cfu.After three cycles of panning,PCR of plasmid DNA prepared from 15 individual phage clones showed that the recombination rate increased from 93% to 100%.BstN1 fingerprinting of insert DNA showed that the diversity of clones decreased with increasing rounds of selection.After three rounds of selection,3 clones showed an identical restriction enzyme pattern.There was a 330-fold enrichment of library phage after 2 rounds of selection and after 3 rounds,a further 8-fold enrichment of library phage was obtained.Conclusion: The ScFv phage display library against human umbilical cord mesenchymal stem cells was successfully constructed.It can be used for succeeding screening of specific antibody against human umbilical cord mesenchymal stem cells and further studying of the cell surface molecules of mesenchymal stem cells.

Objective To explore the feasibility of percutaneous recanalization by retrograde approach via epicardial collaterals. Methods Retrograde percutaneous coronary intervention (PCI) via epicardial collaterals was performed in 5 patients with previously failed antegrade PCI from April 2009 to November 2009. 7 F guiding catheters were engaged in donor artery. Hydrophilic wires and microcathethers were crossed to the distal ends of chronic total occlusion (CTO) lesions via epicardial collaterals. Four retrograde wires were exchanged into stiffer wires and further crossed the CTO, eventually went into the 6 F antegrade guiding catheters and were jailed by a 2.5 mm balloon. After dilatations of retrograde balloons, the lesions were crossed by antegrade wires, and finalized by conventional PCI method. One case was recanalized with retrograde wire trapping technique and another case was recanalized by reverse CART technique. Results The epicardial collaterals were reached from left anterior descending branch (LAD) to distal right coronary artery( RCA ) via apex in 3 patients, from left circumflex branch via left atrium branch to posterior descending artery and RCA in 1 patient and from obtuse marginal artery to diagonal artery and LAD in 1 patient. CTO was successfully recanalized and stents were implanted in 4 patients and failed in 1 patient despite successful wire positioning to the distal end of CTO. There was no procedure-induced cardiovascular event in all cases. Conclusions Epicardial collaterals may not be used as a routine route in retrograde approach PCI due to the potential risk of myocardial rupture and pericardial tamponade. In some cases with unavailable or unsuitable septal collaterals, epicardial collaterals may be used as an alternative route for CTO recanalization.

OBJECTIVETo evaluate the clinical application value of flexible multi-analyte profiling (xMAP) technology in detecting high-risk human papillomavirus (HR-HPV).

METHODSTotally 1 061 women, aged 21-65 years, were randomly enrolled into the study. Cervical exfoliated cells were used in xMAP technology and hybrid capture II (hc2). Pathological diagnosis was used as golden standard. Consistency of these two methods was assessed.

RESULTSThe sensitivity and specificity of xMAP technology were 80.31% and 85.83%, respectively. The positive and negative predictive values were 44.5% and 96.9%, respectively. The Kappa value for consistency between xMAP technology and hc2 was 0.58.

CONCLUSIONSThe specificity of xMAP technology is similar to hc2 test, but the sensitivity is inferior to hc2. However, these two methods show good consistency in the detection of HR-HPV.

Adult ; Aged ; Cervix Uteri ; virology ; Female ; Humans ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; virology ; Sensitivity and Specificity ; Uterine Cervical Diseases ; diagnosis ; virology ; Virology ; methods ; Young Adult

Animals ; DNA, Complementary ; immunology ; Dendritic Cells ; immunology ; Female ; Liver Neoplasms, Experimental ; immunology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; alpha-Fetoproteins ; immunology

The study was purposed to investigate the synergistic reversal effect of Chinese medicine compound FFJZ in combination with cyclosporine A (CsA) on the multidrug resistance (MDR) of human leukemia K562/VCR cell line, as to search effective combination of MDR modulators. MTT (methyl-thazol-tetrazolinum) assay were used to determine the cytotoic and reversal effects on K562/VCR cell line, FCM (flow cytometry) was used to assess the intracellular adriamycin (ADM) concentration and the expression of P-gp in cells. The results showed that the FFJZ in combination with CsA could reverse the drug-resistance of K562/VCR cells and increase the sensitivity K562/VCR cells to adriamycin. They had not the toxic effect on the K562/VCR cells in effective dose and no significant influence on P-gp positive rate of the K562/VCR cells. It is concluded that the FFJZ in combination with CsA may become a safe and effective multidrug resistance-reversing agent with low toxicity in leukemia chemotherapy.
Cyclosporine ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Humans ; K562 Cells ; Vincristine ; pharmacology

OBJECTIVEAdenovirus (ADV) is one of the most common causes of acute respiratory infections in infants and children. The objective of this study was to investigate the prevalence of adenovirus infection among pediatric patients with acute respiratory infections in Beijing and the types of the adenoviruses circulating in Beijing on the molecular bases.

METHODClinical specimens including throat swabs from outpatients and nasopharyngeal aspirates from hospitalized patients were collected from patients with acute respiratory infections in a consecutive period of 6 years from Jan 2003 to Dec 2008. Adenoviruses were identified from the collected clinical specimens by tissue culture and/or immunofluorescence assay and typed by nested-PCR based on the sequence of the encoding gene of hexon. Primers were designed for PCR amplification using hexon gene of adenovirus as target. One primer pair was designed as universal primers for amplifying a 1278 bp gene fragment located at the hexon gene of adenovirus types 3, 7, 11 and 21. Four primer pairs with the sequences located within the region of this 1278 bp fragment were designed specifically for amplifying adenoviruses types 3, 7, 11 or 21, respectively, which were used for a multiplex nest-PCR in a single tube. The products from this multiplex nest-PCR were 502 bp (for type 3), 311 bp (for type 7), 880 bp (for type 11) and 237 bp (for type 21), respectively, and the type of the adenovirus tested can be determined after agarose electrophoresis analysis of the PCR products. For those strains which could not be typed by the multiplex nest-PCR, the gene fragment was amplified by a universal primer pair for all adenovirus types from group A to F and the PCR products were sequenced directly.

RESULTOut of 17 941 clinical specimens collected, including 4378 throat swabs from outpatients and 13 563 nasopharyngeal aspirates from hospitalized patients, 304 were adenovirus positive by tissue culture and/or immunofluorescence assay, the overall positive rate was 1.69% (304/179 41). Among these 304 adenovirus positive specimens, 184 were by virus isolation and 184 by immunofluorescence assay, among which 64 were positive by both methods. The types of the adenoviruses were tested for 285 patients including 174 viral isolates and 111 clinical specimens. By using the multiplex nest-PCR, 272 were typable, including 174 (61.1%, 174/285) for ADV3, 92 (32.3%, 92/285) for ADV7, 6 for ADV11 (2.1%, 6/285) and no adenovirus type 21 was detected. Sequence analysis for those 13 nontypable specimens by the multiplex nest-PCR showed that 9 were ADV2 (3.2%, 9/285), 2 were ADV6 (0.7%, 2/285), 1 was ADV1 (0.4%, 1/285) and 1 was ADV5 (0.4%, 1/285). Most of the patients positive for adenovirus were under 5 years of age and 64.4% were from patients with lower respiratory infections, such as bronchiolitis and pneumonia. All the 5 cases of severe pneumonia with pulmonary failure were caused by ADV7 infection.

CONCLUSIONAdenovirus is still an important pathogen for acute respiratory infection in infants and young children and most of the adenoviruses associated with acute respiratory infections in children in Beijing from 2003 to 2008 were ADV3 and ADV7. ADV7 could cause severe lower respiratory infections.

Acute Disease ; Adenoviridae ; classification ; isolation & purification ; Adenoviridae Infections ; epidemiology ; prevention & control ; Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Prevalence ; Respiratory Tract Infections ; epidemiology ; prevention & control ; virology

OBJECTIVETo study the extraction system of hirudin emulsion liquid membrane with the Poecilobdella manillensis as raw material, di-(2-ethylhexyl) phosphate (D2EHPA) as carrier, Span 80 as emulsifier, octane and D2EHPA mixed to constitute membrane solution, diluted HCl solutions as internal aqueous phase.

METHODUsing the orthogonal experiment to optimize the extraction conditions of hirudin reference substance such as membrane phase, internal aqueous phase volume ratio (MIPVR), external aqueous phase pH, internal aqueous phase pH, mobile carrier concentration and so on, and then using hirudin crude extracts to do purifying experiment, and gaining experimental samples.

RESULTThe optimal conditions of hirudin extraction were as follows: MIPVR 10: 3, internal aqueous phase pH 2.6, external aqueous phase pH 3.4, the mass fraction of carrier D2EHPA 2%. In the optimal extraction conditions, when the initial concentration of hirudin was one anti-thrombin activity units (ATU) x mL(-1), ATU recovery rate of the reference substance was 83.06%. In the purifying experiment of crude extracts, ATU recovery rate was 82.99%, and the specific activity of sample was 3 289.48 the ATU x mg(-1). Discontinuous polyacrylamide gel electrophoresis and spectral scanning, the results showed that the purity and reference substance were considerable.

CONCLUSIONThe method of preparation hirudin was relatively simple, the purity of the experimental samples and ATU recovery were both high.

Animals ; Emulsions ; chemistry ; Hirudins ; analysis ; isolation & purification ; Leeches ; chemistry ; Membranes, Artificial ; Solid Phase Extraction ; instrumentation ; methods