Preparation of Furo[3,2-c]coumarins from 3-Cinnamoyl-4-hydroxy-2H-chromen-2-ones and Acyl Chlorides: A Bu3P-Mediated C-Acylation/Cyclization Sequence.

A Bu3P-mediated cyclization reaction of 3-cinnamoyl-4-hydroxy-2H-chromen-2-ones though electrophilic addition of acyl chlorides towards the synthesis of highly functionalized furo[3,2-c]coumarins bearing a phosphorus ylide moiety is described. These unprecedented cyclization reaction proceeds under mild reaction conditions within short reaction times (1 min to 1 h), and can be further applied in the synthesis of alkenyl-substituted furo[3,2-c]coumarins by the treatment with carbonyl electrophiles under basic conditions.

Syntheses of furo[3,4-c]coumarins and related furyl coumarin derivatives via intramolecular Wittig reactions.

A new and general strategy for highly functional furo[3,4-c]coumarins and related furyl coumarin derivatives has been developed, which is based on an extraordinarily facile intramolecular Wittig reaction, starting from α,ß-unsaturated ketones, tributylphosphine, and acyl chlorides. The phosphorus ylides were proposed to be the key intermediates for constructing the crucial furan ring, leading to a wide variety of substituted furyl coumarins in one step.

[Effects of medicated thread moxibustion of Zhuang medicine on sex hormone in ovariectomized rabbits].

OBJECTIVE: To explore the mechanism of medicated thread moxibustion of Zhuang medicine (MTMZ) in treating perimenopausal period syndrome. METHODS: Thirty rabbits were randomly divided into a normal group, a model group, a medicated thread group, a no-medicated thread group and a sham operation group, 6 cases in each group. The model of perimenopausal period syndrome was established by ovariectomizing the ovary. The medicated thread group was treated with MTMZ at "Qizhou" acupoint (Extra), "Xiaguanyuan"(Extra), "Shenshu" (BL 23) and "Pishu" (BL 20), etc., once each day for 4 weeks. The no-medicated thread group was treated with no-medicated thread moxibustion at the same acupoints, and there is no treatment in the other groups. The changes of hormone level in each group before and after the treatment were observed. RESULTS: After ovariectomizing the ovary, the serum estradiol (E2) in the model group [(308.33 +/- 12.58) pmol/L], the medicated thread group [(304.96 +/- 13.85) pmol/L] and the no-medicated thread group [(303.43 +/- 10.57) pmol/L] were lower than that in the normal group [(478.09 +/- 12.23) pmol/L] and the sham operation group [(488.05 +/- 11.45) pmol/L] (all P < 0.01). After treatment, the E2 level in medicated thread group [(338.92 +/- 11.23) pmol/L] was higher than before (P < 0.01) and that in the model group [(300.53 +/- 13.68) pmol/L] and the no-medicated thread group [(309.74 +/- 13.59) pmol/L] (both P < 0.01), and the serum follicle stimulating hormone [FSH, (58.90 +/- 5.29) U/L] and luteinizing hormone [LH, (64.65 +/- 5.23) U/L] were lower than those in the model group [(65.41 +/- 5.19) U/L], [(71.85 +/- 5.30) U/L] (both P < 0.05). CONCLUSION: MTMZ can increase the serum E2 and reduce the serum FSH and LH in ovariectomized rabbits, and this may be one of the mechanisms of MTMZ for treatment of perimenopausal period syndrome.

Human monoclonal antibody GNX-8 directed to extended type 1 chain: Specific binding to human colorectal cancer.

We observed previously that two carbohydrate epitopes, extended type 1 chain Le(a)-Le(a) and Le(b)-Le(a), are expressed strongly in human gastric or colorectal cancer and cell lines derived therefrom, but their expression in human normal colorectal cells is highly limited. A monoclonal antibody, termed GNX-8, was established through immunization of "KM mice" with colonic cancer cell line Colo205, and with purified Le(b)-Le(a) glycosphingolipid, followed by screening human IgG directed to this antigen. KM mice possess human chromosome fragments and are capable of producing human immunoglobulin. GNX-8 reacted specifically with extended type 1 chain epitope Le(b)-Le(a), bound to all five colonic cancer cell lines so far tested, and displayed strong complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC). The antigens defined by GNX-8, expressed in Colo205 cells, were: (i) glycosphingolipids with epitope Le(b)-Le(a), whose reactivity was abolished upon defucosylation; (ii) glycoproteins with molecular mass range from 32 to >175 kDa, which were depleted in cells cultured in the presence of benzyl-alpha-GalNAc, indicating that these epitopes are O-linked glycans.Immunohistological reactivity of GNX-8 at 1 mug/ml, applied on tissue sections from colorectal and various other types of cancer, was much stronger than that with various normal cells and tissues. GNX-8 reactivity with normal cells required a much higher concentration (150 mug/ml), and this reactivity was based on cross-reaction with non-extended, normal blood group Le(b) antigen. Growth of subcutaneous xenograft of human colonic cancer cells, Colo205 or DLD-1, in nude mice or SCID mice, was strongly inhibited by administration of GNX-8. These observations, taken together, indicate that antibody GNX-8, directed specifically to Le(b)-Le(a) antigen, provides a novel direction of immunotherapy for human colorectal cancer. (c) 2009 UICC.

Development of disposable lipid biosensor for the determination of total cholesterol.

A prototype chronoamperometric biosensor for the determination of total cholesterol was developed that consists of a homemade potentiostat and disposable strips immobilized with Fe(3)O(4), cholesterol oxidase (ChOx), and cholesterol esterase (ChE). The principle of sensing cholesterol is based on the detection of reduction signal of hydrogen peroxide generated in two enzymatic reactions. The co-immobilization of ChE and ChOx allows the sensor to detect both concentrations of esterified and free cholesterol. The effects of biosensor on catalyst, enzymes, applied potential, and buffer pH was investigated, and the operation conditions were optimized. The detection of cholesterol can be accomplished in one step, a 10 microL of sample was dropped onto the area of sensing strip and the reduction signal was obtained at an applied potential of -200 mV (vs. Ag/Ag(+)). The pre-reaction time was set at 15s before applying potential on the strip and the sampling time was 5s. The sensing device displays a linear response over the range of 100-400mg/dL (R(2)=0.999) for cholesteryl oleate. The coefficient variation was determined as 5.06% (N=20) for 100mg/dL cholesteryl oleate and the detection limit is 19.4 mg/dL (S/N=3). The probable interferences in bio-matrix were selected to test the selectivity and no significant response was observed in the biosensor.

Production of antibodies for selective detection of malachite green and the related triphenylmethane dyes in fish and fishpond water.

This study provides a practical method for production of the antibodies against malachite green (MG) and its primary metabolite leucomalachite green (LMG). Two ELISA kits are constructed with the MG and LMG antibodies for detection of the residual MG and LMG in fish muscle and fishpond water. The detection limit is established at the level of 0.05 microg/L for both MG and LMG. Our ELISA kits show the advantages of good specificity, high sensitivity, and convenience in rapid screening of MG and LMG residues. The sample of fishpond water, without extraction or prior preparation, is directly assayed by the ELISA kit. More then 80 fish samples can be simultaneously tested in a kit. The toxic crystal violet and its metabolite leucocrystal violet of illegal use in aquaculture are detected by our prepared MG and LMG antibodies, whereas the antibodies do not cross-react with common antibiotics, sulfonamides, and benzene derivatives.

[Study on RAPD marker linked to sex in Siraitia grosvenorii].

RAPD (Random Amolified Polymorphism DNA) was employed to detect molecular markers linked to sex in S. grosvenorii by BSA (Bulked Segregant Analysis). 18 RAPD markers linked to sex were selected in BSA by screening 90 primers. Only the marker amplified by S1431 was present in all 8 male individuals tested while absent from all 8 female individuals tested. It showed that S1431 was a male RAPD marker linked to sex.