AIM The effects of calcium antagonist diltiazem on plasma insulin levels were investigated in rats. METHODS Plasma insulin was measured with radioimmunoassay method. RESULTS In the fasting rat, plasma insulin levels were significantly increased after ig diltiazem 10 mg?kg -1 and 100 mg?kg -1 , and the level of plasma insulin was increased by 131% 30 min after administration of diltiazem 100 mg?kg -1 in comparison with that of control group. Diltiazem increased the plasma insulin level markedly in glucose tolerance test of the rat. In the hyperglycemia rat induced by adrenaline, diltiazem 100 mg?kg -1 weakly increased the plasma insulin level only at 1 h after administration. CONCLUSION Diltiazem increases the plasma insulin levels in the fasting rat and glucose loaded rat. The increase of plasma insulin level induced by diltiazem is due to the hyperglycemic action of diltiazem and other effect, probably a direct or an indirect insulin release action on pancreas of the rat.

Objective To identify the metabolites of loganin in rat urine through using ultra high performance liquid chromatography combined with triple TOF mass spectrometry (UPLC-Q-TOF/MS/MS) after oral administration of loganin;To summarize the metabolic pathway of loganin. Methods Rats were given a gavage with loganin. The urine samples were prepared by use of solid phase extraction (SPE). The chromatographic separation was performed on an UPLC C18 column with gradient elution program of acetonitrile and 0.1% formic acid aqueous as mobile phase. Dynamic background subtraction (DBS) technology was employed to trigger information dependent acquisition (IDA) to obtain the characteristic fragment ions of metabolites. Results MetabolitePilot software including several data processing methods were employed to speculate the structures of liganin metabolites and three metabolites were identified. Then, the metabolic pathway of loganin in rats was summarized. Conclusion The analytical method was simple, credible and highly-sensible, which is useful for screening and identifying trace constituents in vivo.

Complicated characteristic of facial modality acquisition and measurement is commonly needed in clinical environment.Two different methods were compared, one method was acquisition two-dimensional character based on the principle of machine vision with a charge-coupled device (CCD) capturing image, and another method was acquisition three-dimensional character based on the technology of Reverse Engineering with laser scanning capturing image. The principle, composition of the system, calibration and characteristics of different method are analyzed. Two-dimensional and three-dimensional facial data of thirty undergraduates was used to compare different methods. The application and restriction of different methods are concluded. The conclusion of this paper provided a reference for face plastic surgery, facial paralysis, facial surgical evaluation and rehabilitation design.

Objective To analyze the outcome of patients receiving intrauterine insemination with husband sperm,in order to evaluate the effect of relative factors on pregnancy rate after intrauterine insemination. Methods Ninety-eight infertile couples who received intrauterine insemination in the Affiliated Hospital of Nantong University from March 2013 to May 2014 were selected as our subjects and 181 cycles were included. The information including clinical factors including maternal age,infertile time,infertile causes, ovulation induction protocol,time of insemination and postwash total motitle sperm(TMS)and pregnancy rate were recorded. Results (1)Totally 26 patients received clinical pregnancies,and clinical pregnancy rate(CPR) was 14. 36% per cycle. With age increase pregnancy rate decreased( χ2 = 1. 654 9,P = 0. 647).(2)The pregnancy rate of the patients was the same within the infertile time( χ2 = 1. 588 5,P = 0. 662).(3)The pregnancy rate of the patients with secondary infertility was lower than that of the patients with primary infertility,but there was no significant difference(χ2 = 0. 923 3,P = 0. 337).(4)The pregnancy rate of ovulation induction cycles was lower than that of nature cycles,but there was no significant difference(χ2 = 2. 222 0,P= 0. 136).(5)Postwash TMS was showed the same trend(χ2 = 0. 643 4,P = 0. 422). Conclusion In terms of intrauterine insemination with husband sperm,age,infertile time,infertile types,ovulation induction protocol and posrwash TMS can affect pregnancy rate,and the effects of various factors should be considered comprehensively in the process of therapy.

Objective To study the application of dynamic ultrasound countercurrent extraction technology in industrialization of Lianhua Qingwen Capsule by multi-index optimization. Methods Ultrasonic temperature and ultrasonic time were investigated by using single factor design. The three factors of dry extract yield, the active ingredient content, and fingerprints were selected as the optimization indexes to investigate the solvent consumption and extraction time between the dynamic ultrasound countercurrent extraction process and the original reflux extraction process under the fixed conditions of process parameters. Results Compared with traditional reflux extraction technology, dynamic ultrasound countercurrent extraction technology effectively reduced the alcohol consumption and saved extraction time on the basis of guarantee of the active ingredient content. Conclusion Dynamic ultrasound countercurrent extraction process, with simple and fast advantages, can be used to Lianhua Qingwen Capsule production, which can significantly reduce production costs and increase productivity.

Objective: Expression of Bcl\|2 and Bax oncoproteins was tested in laryngeal squamous cell carcinoma (LSCC) in order to find the relationship between the two genes and LSCC, and to explrore their clinical significance by retrospectively study of pathological and clinical doouments. Methods: Immunohistochemical S\|P method was emioyed in testing the expression of Bcl\|2 and Bax oncoproteins in 74 patients with LSCC, 24 laryngeal atypical hyperplasis (LAH) and 24 laryngeal normal membrane (LNM). Results: Bcl\|2 oncoprotein was mainly expressed in cytoplasm of epithelium cells and a small part of cell membrances were stained, appearing brow or yellow, granular distribution.There were no expression in the cell nuclear and stroma. Bax oncoprotein was expressed in cytoplasm and cell membrane and there were a little expression in stroma, appearing light brown and yellow, granular distribution. Bax was less stained than Bcl\|2. The positive expression percentages of Bcl\|2 oncoprotein in LSCC, LAH, LNM tissues were 59.46%,66.67% and 20.83% respectively. The Bcl\|2 expressions in LSCC, LAH tissues were significantly higher than that in LNM (P＜0.05); The positive expression percentages of Bax oncoprotein in these three groups were 56.76%, 50.0% and 66.67% respectively which were no significant difference. The results of Spearman correlated analysis showed that expression of Bcl\|2 and Bax oncoproteins had no obvious correlations with pathological grade, clinical stage and metastasis of lymph node. Both were correlated only with smoking. Conclusions: The imbalance of Bcl\|2/Bax mainly induced by the high expression of Bcl\|2 oncoprotein played an important role in the development of LSCC and LAH, Bcl\|2 may act as a reference index in judging the biological speciality of LSCC; Expressions of Bcl\|2 and Bax oncoprotein had obvious corelation with smoking in LSCC, indicating Bcl\|2 and Bax genes may be a target of carcinogenic substance in tobacco; Expression of Bcl\|2 oncoprotein was blocked may be an important part in studying gene therapy for the future.

OBJECTIVE:To establish a method for simultaneous determination of calycosin-7-glucoside,specnuezhenide,ber-berine hydrochloride,palmatine hydrochloride and rosmarinic acid in Xialiqi capsule. METHODS:UPLC was performed on the col-umn of Shield RP18 with mobile phase of nitrile-0.2%formic acid(gradient elution)at a flow rate of 0.2 ml/min,the column tem-perature was 35 ℃,the detection wavelength was 240 nm,and the injection volume was 1 μl. RESULTS:The linear range was 0.619-39.6 μg/ml for calycosin-7-glucoside(r=0.999 9),1.181-115.6 μg/ml for specnuezhenide(r=0.999 9),0.89-57.2 μg/ml for berberine hydrochloride(r=0.999 9),1.16-74 μg/ml for palmatine hydrochloride(r=0.999 9)and 2.47-157.8 μg/ml for rosmarinic acid(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 3%,recoveries were 97.98%-102.44%(RSD=1.49%,n=9),97.37%-100.96%(RSD=1.13%,n=9),98.53%-102.30%(RSD=1.40%,n=9),97.06%-102.21%(RSD=1.46%,n=9)and 100.10%-102.25%(RSD=0.68%,n=9),respectively. CONCLUSIONS:The method is simple,rapid and reproducible,and can be used for the quality control of Xialiqi capsule.

Objective To solve the agglomeration problem in the former process, dry granulation technology was used to prepare the granules of Lianhua Qingwen Capsules. Methods Complexity and granule yield coefficient were set as inspection indexes. The optimum subsidiary material and its amount were optimized. The parameters of dry granulation technology were optimized by orthogonal test. Then, granule yield, angle of repose, and bulk density were compared with those of wet granulation technology. Results Starch was set as subsidiary material. The optimum technology is roll pressure of 12 MPa, rotation speed of 5 r/min, and feed speed of 10 r/min. The yield of granules prepared by dry granulation technology was significantly higher than that of wet granulation technology. Conclusion The dry granulation technology can effectively improve the agglomeration of the granulation process of Lianhua Qingwen Capsules, and was suitable for granulating process of Lianhua Qingwen Capsules.

BACKGROUND:CTLA-4Ig as a tolerance-induction agent is a potential strategy in graft-versus-host disease prevention. OBJECTIVE:To investigate the efficacy of CTLA4Ig-gene-modified bone marrow stromal cels mediated by adenovirus to induce T-cel tolerance of haploidentical donors. METHODS: The bone marrow stromal cels isolated culture from the bone marrow of HLA haploidentical donors were transfected by recombinant adenovirus encoding CTLA4IgcDNA (AdCTLA4Ig) at a multiplicity of infection=50 for 72 hours. The expression rate and the location of CTLA4Ig in the transfected cels were detected by fluorescence microscope after immunofluorescence staining. CTLA4Ig-modified bone marrow stromal cels (2×104, 4×104and 8×104) were respectively co-cultured with 105 T cels from the peripheral blood of HLA haploidentical donors and 105 peripheral blood mononuclear cels from recipients. The proliferative inhibition rate was determined by MTT assay, and the level of interleukin-2 in the supernatant was detected by ELISA. The bone marrow mononuclear cels (1×105/wel) were co-cultured with CTLA4Ig-modified bone marrow stromal cel layers constructed in 6-wel plates. The number of bone marrow mononuclear cels and colony-forming unit-granulocyte macrophages were calculated after 5-day culture. RESULTS AND CONCLUSION: The expression rate of CTLA4Ig at the multiplicity of infection=50 was as high as 85%, and the immunofluorescence signals of CTLA4Ig were distributed unevenly in the cytoplasm. The inhibition rates of 2×104, 4×104, and 8×104 CTLA4Ig-modified bone marrow stromal cels on proliferation of T cels were higher than that of untransfected cels. The levels of interluekin-2 in the corresponding cel groups were significantly lower than that in the untransfected cels (P < 0.05). At 5 days of culture, there was no significant difference in the number of bone marrow mononuclear cels and colony-forming unit-granulocyte macrophages between the transfected and untransfected cel groups (P > 0.05). These findings indicate that CTLA4Ig-modified bone marrow stromal cels mediated by adenovirus can induce immune tolerance of T-lymphocyte from HLA haploidentical donors in vitro.