OBJECTIVE: To establish methods for screening of macromolecules in traditional Chinese medicine injections by molecular exclusion chromatography (SEC). METHODS: The separation was carried out by SEC with Phenomenon BioSep-SEC-s2000 (7.8 mm×300 mm, 8 μm) column. The mobile phase was 0.05 mol·L-1 Na2SO4, and differential detector was used when dextran was used as the reference standard. When proteins were used as reference standards, the mobile phase was the mixture of 0.1% trifluoroacetic acid and acetonitrile (70-30), and UV detector was selected. RESULTS: With the differential detector, the molecular weight range of dextran was 7100-133800, and the limit of detection was 0.0979 μg; with the UV detector, the molecular weight range of the proteins was 1638-13700, and the limit of detection was 0.00358 μg. No macromolecules were detected in the selected traditional Chinese medicine injections. CONCLUSION: The proposed method can be used for the screening of macromolecules in traditional Chinese medicine injections.

AIM: To investigate the preoperative binocular visual function of intermittent exotropia and the rebuilding and recovery of the postoperative binocular visual function, and analyze the effect of binocular visual function on orthophoria after surgery.
METHODS:From January 2011 to January 2014, 47 basic intermittent exotropia patients caming for treatment were collected in the clinical data. The changes in their near stereopsis, binocular visual function, binocular fusion and distance stereopsis after operations were recorded in the form of data. The preoperative binocular vision and the postoperative rebuilding were analyzed and contrasted with each other. In addition, the effect on the postoperative maintaining of orthophoria due to the existence, recovery and rebuilding of binocular visual function were observed.
RESULTS:Intermittent exotropia patients got damage in different levels on their binocular visual functions, especially on distance stereopsis, which was the heaviest and earliest. After the operation, all functions were obviously recovered and reconstructed and the improvements were statistically significant compared against those before the operation (P<0. 01). Patients having binocular visual function or part of it before the operation had a higher ratio of orthophoria compared against the patients who had lost binocular visual function before the operation and the difference was statistically significant (P<0. 01). The recovery and reconstruction of the postoperative binocular visual function played an important role in maintaining the orthophoria.
CONCLUSION: The intermittent exotropia cause damage to the stereopsis which happened the earliest. Obvious recovery and reconstruction of binocular visual function can be observed after the surgery. A relatively good preoperative binocular visual function may lead to the increase in the ratio of orthophoria or cure the intermittent exotropia. Performing an operation when distance stereopsis is damaged can increase the success rate for the surgery and reduce the recurrence rate.

Objective To establish a standardized and feasible family health assessment index system based on the family practice system in Shanghai urban communities.Methods On the basis of literature review and special topic discussion, a preliminary index system framework was developed and an expert consultation questionnaire was compiled.Twenty one experts of general practice, medical specialties, health administration and public health management were invited and two rounds of improved Delphi expert consultation were conducted.The results of statistical analysis were summarized and the evaluation index system was established.Results The evaluation index system included 5 first-level indicators, 17 second-level indicators and 63 third-level indicators.The weight coefficients of first-level indicators in basic information, family structure, family function, family resources and health status of the family members were 0.199 6, 0.197 5, 0.201 7, 0.185 0, 0.216 2, respectively.In the two rounds of consultation, the positive coefficient of experts was both 100%, the degree of expert authority was 0.867 and 0.880, and the coordination coefficient was 0.294 and 0.415, respectively (P<0.05).The operational coordination coefficients were 0.320 and 0.402 (P<0.05), respectively.Conclusion The family health assessment index system constructed by the Delphi expert consultation method can provide reference for family doctors to carry out family health assessment in urban communities.

Objective To evaluate the performance of actinic keratosis (AK) tissue as a culture model for the study of interference in transduction pathway,and to explore the mechanism underlying the p53 regulation though TGFβ1/Smads pathway by using the tissue culture model.Methods Twenty-five skin samples from the lesions of patients with AK were cultured,and divided into 5 groups to be treated with TGFβ1 of 10 μg/L for 24 and 48 hours,the tran sforming growth factor (TGF) β1 receptor kinase inhibitor SB431542 of 10 μmol/L for 24 and 48 hours,respectively,or remain untreated.Real time PCR and Western blot were performed to quantify the mRNA expression of p53 and protein expression of p53 and phosphorylated Smad2 in these tissue specimens respectively.Results A significant elevation was observed in the expressions of p53 mRNA ( 13.4968 ± 0.9903 vs.1,P ＜ 0.05) and phosphorylated Smad2 (0.700 ± 0.023 vs.1,P ＜ 0.05) in AK tissues after treatment with TGFβ1 for 24 hours,and in the expressions of p53 mRNA (13.3882 ± 1.6772 vs.1,P ＜ 0.05) and protein (1.009 ± 0.001 vs.0.512 ± 0.005,P ＜ 0.05) after treatment with TGFβ1 for 48 hours,compared with the untreated AK tissues.No significant differences were observed in the expression of p53 protein between the AK tissues treated with TGFβ1 for 24 hours and 48 hours (P ＞ 0.05).SB431542 induced a statistical reduction in the level of phosphorylated Smad2 at 48 hours (0.116 ± 0.003 vs.0.306 ± 0.023,P ＜ 0.05),but no significant changes were observed in the expression of p53 mRNA or protein after SB431542 treatment for 24 or 48 hours.Conclusions AK tissue cultures can serve as a model for the study of interference in signal transduction pathway.TGFβ1 might regulate the expression of p53 protien through Smads pathway in AK.

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

The study aims to develop a unified method to determine seven phenolic acids (neochlorogenic acid, chlorogenic acid, 4-caffeoylquinic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C) contained in honeysuckle flower that is the monarch drug of all the eight Yinqiao Jiedu serial preparations using quantitative analysis of multi-components by single-marker (QAMS). Firstly, chlorogenic acid was used as a reference to get the average relative correction factors (RCFs) of the other phenolic acids in ratios to the reference; columns and instruments from different companies were used to validate the durability of the achieved RCFs in different levels of standard solutions; and honeysuckle flower extract was used as the reference substance to fix the positions of chromatographic peaks. Secondly, the contents of seven phenolic acids in eight different Yinqiao Jiedu serial preparations samples were calculated based on the RCFs durability. Finally, the quantitative results were compared between QAMS and the external standard (ES) method. The results have showed that the durability of the achieved RCFs is good (RSD during 0.80% - 2.56%), and there are no differences between the quantitative results of QAMS and ES (the relative average deviation < 0.93%). So it can be successfully used to the quantitative control of honeysuckle flower principally prescribed in Yinqiao Jiedu serial preparations.
Caffeic Acids ; analysis ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Flowers ; chemistry ; Hydroxybenzoates ; analysis ; Lonicera ; chemistry ; Quinic Acid ; analogs & derivatives ; analysis

A study on the microbial transformation of glycyrrhetinic acid (GA) was conducted by a fungus, Cunninghamella blakesleeana CGMCC 3.970 systematically. After incubation with the cell cultures of C. blakesleeana CGMCC 3.970 at 25 degrees C for 7 days on a rotary shaker operating at 135 r x min(-1), GA was converted into one major product and five minor products. The products were extracted and purified by solvent extraction, macroporous adsorbent resin, silica gel column chromatography, and semi-preparative RP-HPLC chromatography. Their structures were identified as 3-oxo-15α-hydroxy-18β-glycyrrhetinic acid(1), 3-oxo-15β-hydroxy-18β-glycyrrhetinic acid (2), 7β,15α-dihydroxy-18β-glycyrrhetinic acid (3), 3-oxo-7β, 15α-dihydroxy-18β-glycyrrhetinic acid (4), 7β-hydroxy-18β-glycyrrhetinic acid(5) and 15α-hydroxy-18β-glycyrrhetinic acid(6) by the analyses of MS, 1H-NMR and 13C-NMR spectroscopic data respectively. Among them, 2 was a new compound. These results suggest that C. blakesleeana CGMCC 3.970 has the capability of selective ketonization and hydroxylation for GA. [Key words] glycyrrhetinic acid; Cunninghamella blakesleeana CGMCC 3. 970; microbial transformation
Biotransformation ; Cunninghamella ; metabolism ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; metabolism ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Objective To estimate the effect of different doses of ultraviolet (UV) radiation on the proliferation of and apoptosis in kertatinocytes,as well as on the expression of p53,matrix metalloproteinase-2 (MMP2) and-9 (MMP9) in actinic keratosis (AK) lesions and normal human skin.Methods Tissue specimens were obtained from the lesions of 20 patients with AK and sun-exposed normal skin of 20 healthy human subjects,and subjected to an air-exposed culture.Each of the specimens was divided into 4 areas to remain untreated (control area) or be irradiated with UV of 5,10 and 20 J/cm2 (irradiated areas) for 4 consecutive days.After another 24-hour culture,the tissue cultures were collected followed by the evaluation of apoptosis in and proliferation of keratinocytes by using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and Ki-67 staining,and determination of mRNA and protein expressions of p53,MMP2 and MMP9 by using real time PCR and immunohistochemistry respectively.Results A statistical increase was observed in the percentage of apoptotic cells in the normal skin irradiated with UV of 10 and 20 J/cm2 (46.8％ ± 2.1％ and 56.7％± 2.4％,both P ＜ 0.05) and in the AK lesions irradiated with UV of 20 J/cm2 (43.5％ ± 1.5％,P ＜ 0.05)compared with the corresponding unirradiated tissues.The normal skin showed a higher percentage of apoptotic cells than the lesional skin after irradiation with UV of 10 and 20 J/cm2 (both P ＜ 0.05).The percentage of Ki67-positive cells was significantly decreased in the normal skin after irradiation with UV of 20 J/cm2 (3.34％ ±0.76％,P ＜ 0.05),but experienced no statistical changes in the lesional skin after different doses of UV irradiation (all P ＞ 0.05).There was a statistical elevation in the expression of p53 mRNA (5 J/cm2:1.106 ± 0.025,10 J/cm2: 1.259 ± 0.045,20 J/cm2:1.425 ± 0.053,all P ＜ 0.05) and protein(10 J/cm2:0.1169 ± 0.0032,20 J/cm2:0.1454 ± 0.0047,both P＜ 0.05) in the normal skin,but a statistical reduction in the expression of p53 mRNA(10 J/cm2.0.611 ± 0.050,20 J/cm2:0.578 ± 0.070,both P ＜ 0.05) and protein (20 J/cm2:0.0404 ± 0.0027,P＜ 0.05) in the lesional skin after irradiation compared with the corresponding unirradiated skin tissues.Further more,a statistical increment was observed in MMP2 mRNA and protein expression in normal skin irradiated with UV of 10 J/cm2 (1.086 ± 0.013,0.0843 ± 0.0024,respectively,both P ＜ 0.05) and 20 J/cm2 (1.417 ± 0.036,0.1236 ±0.0042,respectively,both P ＜ 0.05) and in lesional skin irradiated with UV of 20 J/cm2 (1.296 ± 0.028,0.0744± 0.0032,respectively,both P ＜ 0.05),as well as in MMP9 mRNA and protein expression in normal skin irradiated with UV of 20 J/cm2 (1.395 ± 0.026,0.3065 ± 0.0162,respectively,both P ＜ 0.05) and in lesional skin irradiated with UV of 10 J/cm2 (1.298 ± 0.035,0.0992 ± 0.0053,respectively,both P ＜ 0.05) and 20 J/cm2(1.286 ± 0.032,0.1010 ± 0.0063,respectively,both P ＜ 0.05) compared with the corresponding unirradiated tissues.Conclusion Ultraviolet may accelerate the progression of AK by down-regulating p53 expression but up-regulating MMP2 and MMP9 expression.

Objective To study the protective effect and its mechanism of anti-tumor necrosis factor-?antibody (TNF-? Ab) on lung injury after cardiopulmanary bypass(CPB).Methods Twenty-eight healthy rabbits were selected and randomly divided into four groups:group Ⅰ only received open chest operation;groups Ⅱ-Ⅳ underwent CPB.In the group Ⅳ,rabbit TNF-? Ab (2 400 pg/kg) was dropped into the intracheal tube before operation and just after releasing the aortic clamp.Saline was given to the group Ⅲ in- stead.Blood neutrophils count,TNF-?,MDA from the right and left atrium in the four groups were determined perioperatively.Water volume,TNF-? mRNA,TNF-? protein,apoptosis and pathomorphological changes were measured in the lung tissues.Results TNF- ? Ab can restrain leukocyte accumulation,reduce releasing of TNF-? and MDA in the lung.It can also reduce the occurrence of apop- tosis and attenuate pathomorphological changes in the lung tissue.However,it cannot reduce the secretion of TNF-? at the transcrip- tion level and protein level.Conclusion Intratracheal TNF-? Ab administration has markedly protective effect on lung injury after CPB.

OBJECTIVETo improve the paramedian forehead flap for nasal reconstruction.

METHODSBased on the findings of the Cutaneous branch of supratrochlear artery, the forehead musculo-cutaneous flap was divided into the musculo-flap and the skin-flap in 3 patients. The musculo-flap were used to reconstruct the septi-bone structure, and envelop silicon nasal-frame, while the skin-flap were used to reconstruct the nose. In 6 patients, the forehead flap was used as a skin-flap just with the muscle pedicle to reconstruct a nose with good appearance of nasal subunits, without the secondary operation.

RESULTSIn all of 9 cases, the cutaneous-branch of supratrochlear artery was found 1.7-2.0 cm above the orbital-rim. Eight patients got good results, only 1 patient had to do the secondary operation to coverage the exposed nasal frame.

CONCLUSIONSThe cutaneous-branch of supratrochlear artery could be an anatomical finding to form the forehead skin-flap and it is a good choice to be used to reconstruct a nose, even in complicated cases.

Adolescent ; Adult ; Female ; Forehead ; Humans ; Male ; Muscles ; transplantation ; Nose ; surgery ; Patient Satisfaction ; Reconstructive Surgical Procedures ; methods ; standards ; Skin Transplantation ; methods ; Surgical Flaps ; Treatment Outcome