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.

OBJECTIVEto explore the feasibility of percutaneous recanalization by retrograde approach via epicardial collaterals.

METHODSretrograde 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 microcatheters 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.

RESULTSthe 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.

CONCLUSIONSepicardial 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.

Aged ; Angioplasty, Balloon, Coronary ; methods ; Arteriosclerosis Obliterans ; therapy ; Collateral Circulation ; Coronary Artery Disease ; therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo develop a new technique of bilateral angiography in a single radial access (BASiRalA) which can reduce a puncture site.

METHODSFrom March 2011 to February 2012, 13 cases of coronary heart disease patients with chronic total occlusion (CTO) were treated (6 CTOs in right coronary artery and 7 in left anterior descending artery). All patients underwent percutaneous coronary intervention (PCI) via the right radial artery access and 6 F guiding catheters were delivered to the diseased artery. Once the wires crossed the CTO lesions and were uncertain if the wires were in true lumen or not, BASiRalA was performed. The Finecross microcatheters were advanced out of the 6 F guiding catheter, then withdraw 6F guiding catheter to the opening of diseased artery, the soft wires were manipulated into the middle portion of opposite coronary artery. After that, the microcatheters were advanced to this segment or the branches relative to the collateral vessels connected with CTOs. After pulling out the wires, microcatheter injections can be performed for contralateral angiography. BASiRalA related complications were observed after the procedure.

RESULTSBASiRalA technique was applied to 13 CTOs and 10 procedures succeeded (76.92%). BASiRalA failed in 3 cases and the wires and microcatheters could not be advanced to the opposite coronary arteries within 20 minutes. Alternatively, contralateral angiography via femoral arteries was performed in these 3 patients. The average time of BASiRalA technique was 7 (5 - 13) minutes and the shortest time of wires crossing to the opposite coronary artery was 5 seconds. There was no procedure induced complication during procedure or post procedure.

CONCLUSIONBASiRalA technique is feasible in treating CTO patients by PCI.

Aged ; Angioplasty, Balloon, Coronary ; Cardiac Catheterization ; methods ; Coronary Angiography ; methods ; Coronary Occlusion ; therapy ; Female ; Humans ; Male ; Middle Aged ; Radial Artery ; Retrospective Studies

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

BACKGROUNDSurgical interventions for moyamoya disease include direct and indirect revascularizations. This study aimed to evaluate the therapeutic effect of superficial temporal artery-middle cerebral artery bypass combined with an indirect revascularization procedure, encephalo-duro-myo-synangiosis, in the treatment of moyamoya disease.

METHODSFrom October 2005 to November 2009, we performed this combined revascularization procedure in 111 patients with different types and stages of moyamoya disease. The superficial temporal artery, middle meningeal artery and the deep temporal artery were evaluated for individualized surgical planning in these cases. The integrity of the deep temporal artery and the middle meningeal artery network, and the pre-existing spontaneous anastomoses of the distal branches of the external carotid artery with the cortical arteries were well preserved. The mean follow-up time was 72.5 months, all clinical and radiological data were retrospectively reviewed.

RESULTSA total of 198 stomas were performed in 122 hemispheres, all remaining patent until the last follow-up. The encephalo-duro-myo-synangiosis resulted in extensive anastomoses of the deep temporal artery (100%), the middle meningeal artery (90.9%), and the sphenopalatine artery (39.8%) with the cortical arteries, respectively. The superficial temporal artery, deep temporal artery, and the middle meningeal artery were significantly thickened in 88 patients as determined by digital subtraction angiography at follow-up. The relative cerebral blood flow increased significantly within one week after the operation. At 6 months post the operation, the relative cerebral blood flow was further increased by 15.5% from the gradual formation of anastomoses as a result of indirect revascularization. Transient ischemic attacks were effectively reduced or totally arrested. The neurological deficits significantly improved in 37 patients, with the National Institutes of Health Stroke Scale scores lowered by 2-8. There was no rehemorrhage in hemorrhagic moyamoya disease patients.

CONCLUSIONThis study showed that the superficial temporal artery-middle cerebral artery bypass combined with encephalo-duro-myo-synangiosis can achieve good therapeutic effect in the treatment of moyamoya disease.

Adolescent ; Adult ; Cerebral Revascularization ; methods ; Child ; Female ; Humans ; Male ; Middle Aged ; Middle Cerebral Artery ; pathology ; surgery ; Moyamoya Disease ; pathology ; surgery ; Young Adult

OBJECTIVETo prepare morphine-loaded chitosan microspheres by emulsion ionic cross-linking and investigate the effect of initial morphine quantity and different cross-linking degrees on drug loading, encapsulation efficiency and in vitro drug release.

METHODSChitosan (with a relative molecular mass of 50,000 and deacetylation degree no less than 90%) at 100 mg and morphine at 20, 30, 40, or 50 mg were dissolved by 2% acetate and dripped slowly into 15 ml soy-bean oil containing 0.75 ml Span80. After full emulsification at 35 degrees C; for 1.5 h, the mixture was dripped slowly into sodium tripolyphosphate (10 mg/ml) at the mass ratio of 5:1, 7:1, or 9:1 to allow cross-linking for 2 h. The drug loading, encapsulation efficiency and in vitro drug release of the preparations were measured.

RESULTSThe drug loading in the microsphere increased while the encapsulation efficiency reduced with the increment of the initial morphine quantity. High cross-linking degree resulted in prolonged release time of the drug loaded in the preparations.

CONCLUSIONThe microspheres loaded with morphine allows sustained release of morphine.

Chitosan ; administration & dosage ; Delayed-Action Preparations ; chemical synthesis ; Drug Carriers ; administration & dosage ; Microspheres ; Morphine ; administration & dosage

BACKGROUNDIn humans telomerase is expressed in most cancers and immortal cell lines, and activation of telomerase may play important roles in tumorigenesis and immortalization. This study was to investigate the roles of telomerase activity (TA) and human telomerase RNA (hTR) in sebaceous carcinoma of the eyelid.

METHODSThe telomerase repeated amplification protocol (TRAP) was used to demonstrate telomerase activity in 12 cases of sebaceous carcinoma of the eyelid. In situ hybridization (ISH) was used to demonstrate the expression of hTR in 55 cases of paraffin-embedded sebaceous carcinoma of the eyelid, and the results were compared with the proliferative index determined by Mib-1 immuno-labeling, histological patterns and recurrence of the tumor.

RESULTSDifferent telomerase activity was shown in the 12 cases of sebaceous carcinoma of the eyelid. The positive expression of hTR was 85.5% (47/55) in tumor cells, but not in the adjacent tissues. The positive expression of hTR was correlated with the proliferative activity (as assessed by Mib-1 immunolabelling, r = 0.942, P < 0.001) and the differentiation of sebaceous carcinoma of the eyelid (chi(2) = 17.621, P < 0.001), but not significantly related to tumor recurrence. The level of hTR expression increased with the decrease of differentiation of sebaceous carcinoma of the eyelid.

CONCLUSIONSThe results suggest that the up-regulation of telomerase expression plays some roles in tarsal gland carcinogenesis, and the expression of hTR is a useful marker for malignant degree of sebaceous carcinoma of the eyelid.

Biomarkers, Tumor ; analysis ; Eyelid Neoplasms ; enzymology ; pathology ; Humans ; In Situ Hybridization ; Neoplasm Recurrence, Local ; enzymology ; RNA ; analysis ; Sebaceous Gland Neoplasms ; enzymology ; pathology ; Telomerase ; analysis