Dietary supplementation of proteases on growth performance, nutrient digestibility, blood characteristics and gut microbiota of growing pigs fed sorghum-based diets.

Low-tannin sorghum is an excellent energy source in pig diets. However, sorghum contains several anti-nutritional factors that may have negative effects on nutrient digestibility. The impacts of proteases on growth performance, nutrient digestibility, blood parameters, and gut microbiota of growing pigs fed sorghum-based diets were studied in this study. Ninety-six pigs (20.66 ± 0.65 kg BW) were allocated into three groups (eight pens/group, four pigs/pen): (1) CON (control diet, sorghum-based diet included 66.98% sorghum), (2) PRO1 (CON + 200 mg/kg proteases), (3) PRO2 (CON + 400 mg/kg proteases) for 28 d. No differences were observed in growth performance and apparent total tract digestibility (ATTD) of nutrients between CON and PRO1 groups. Pigs fed PRO2 diet had increased (P < 0.05) BW on d 21 and 28, and increased (P < 0.05) average daily gain during d 14-21 and the overall period compared with pigs fed CON diet. In addition, pigs fed PRO2 diet had improved (P < 0.05) ATTD of gross energy, CP, and DM compared with pigs fed CON and PRO1 diets. Pigs fed PRO2 diet had lower (P < 0.05) plasma globulin (GLB) level and higher (P < 0.05) plasma glucose, albumin (ALB) and immunoglobulin G levels, and ALB/GLB ratio than pigs fed CON and PRO1 diets. Furthermore, pigs fed PRO2 diet had decreased (P < 0.05) the relative abundance of Acidobacteriota at the phylum level and increased (P < 0.05) the relative abundance of Prevotella_9 at the genus level. The linear discriminant analysis effect size analysis also showed that pigs fed PRO2 diet had significantly enriched short-chain fatty acid-producing bacteria, such as Subdoligranulum and Parabacteroides. In conclusion, protease supplementation at 400 mg/kg improved the growth performance of growing pigs fed sorghum-based diets, which may be attributed to the improvement of nutrient digestibility, host metabolism, immune status and associated with the altered gut microbiota profiles.

Effects of dietary methionine supplementation from different sources on growth performance and meat quality of barrows and gilts.

Methionine is indispensable for growth and meat formation in pigs. However, it is still unclear that increasing dietary sulphur-containing amino acid (SAA) levels using different methionine sources affects the growth performance and meat quality of barrows and gilts. To investigate this, 144 pigs (half barrows and half gilts) were fed the control (100% SAA, CON), DL-Methionine (125% SAA, DL-Met)-supplemented, or OH-Methionine (125% SAA, OH-Met)-supplemented diets during the 11-110 kg period. The results showed that plasma methionine levels varied among treatments during the experimental phase, with increased plasma methionine levels observed following increased SAA consumption during the 25-45 kg period. In contrast, pigs fed the DL-Met diet had lower plasma methionine levels than those fed the CON diet (95-110 kg). Additionally, gilts fed the DL-Met or OH-Met diets showed decreased drip loss in longissimus lumborum muscle (LM) compared to CON-fed gilts. OH-Met-fed gilts had higher pH45min values than those fed the CON or DL-Met diets, whereas OH-Met-fed barrows had higher L45min values than those fed the CON or DL-Met diets. Moreover, increased consumption of SAA, regardless of the methionine source, tended to decrease the shear force of the LM in pigs. In conclusion, this study indicates that increasing dietary levels of SAA (+25%) appeared to improve the meat quality of gilts by decreasing drip loss and increasing meat tenderness.

Clinical effectiveness of 3 days preoperative treatment with recombinant human erythropoietin in total knee arthroplasty surgery: a clinical trial.

AIMS: The purpose of study is to evaluate the effect and complication of preoperative short-term daily recombinant human erythropoietin (rhEPO) treatment for blood-saving in patients undergoing unilateral primary total knee arthroplasty (TKA). METHODS: This three-arm randomized clinical trial compared three different rhEPO-based treatment protocols for unilateral primary TKA. Group A: application of daily doses of rhEPO combined with iron supplement starting 3 days before surgery; Group B: application of daily doses of rhEPO combined with iron supplement starting the day of surgery; Group C: iron supplement alone. Perioperative hemoglobin (Hb) level gaps, total perioperative blood loss, reticulocyte levels and treatment-related complications were studied. RESULTS: A total of 102 patients were included (35, 35 and 32 patients in Groups A, B and C, respectively). Total blood loss (TBL) in Groups A, B and C was 490.84, 806.76 and 924.21 ml, respectively. Patients in Group A had a significant lower TBL than Groups B and C (A vs. B: P = 0.010; A vs. C: P < 0.001). There was no difference as for TBL between Groups B and C (P = 0.377). Group A patients had significant smaller Hb decline than Group C on the third and fifth postoperative day (P = 0.049, P = 0.037), as well as than Group B on the fifth postoperative day (P = 0.048). There was no difference as for Hb decline between Groups B and C. No difference was shown in levels of inflammatory biomarkers or blood-saving protocol-related complications among three groups. CONCLUSIONS: Daily dose of rhEPO combined with iron supplement administered 3 days before TKA procedures could significantly decrease perioperative blood loss and improve postoperative Hb levels, without significantly elevating risks of complication, when compared with admission of rhEPO on the day of surgery and iron supplement alone. Preoperative daily rhEPO treatment could be a more effective blood-saving protocol in TKA procedures.

[The status analysis of diagnosis and treatment of synchronous peritoneal carcinomatosis from colorectal cancer in China: a report of 1 003 cases in 16 domestic medical centers].

Objective: To analyze the status of domestic surgical treatment of synchronous peritoneal carcinomatosis from colorectal cancer in China. Methods: Clinicopathological data of patients who underwent surgery from October 2003 to October 2018 in 16 domestic medical centers was retrospectively analyzed. Excel database was created which covered 77 fields of 7 parts: baseline information of patients, laboratory tests, imaging tests, chemoradiotherapy information, intra-operative findings, postoperative pathology and follow-up data. The Wilcoxon rank-sum test was used for comparison of the measurement data between groups. The χ(2) test was used for comparison of the categorical data between groups. The survival curve was calculated by the Kaplan-Meier method. Results: Of the 1 003 patients, there were 575 male and 428 female patients with the age of (58.5±14.1) years (range: 18 to 92 years). In a total of 920 patients, the carcinoma of sigmoid colon was performed in 292 cases (31.8%) with the highest ratio. The proportion of patients with liver metastasis and lung metastasis were 27.9% (219/784) and 8.3% (64/769). Preoperative detection of carcino-embryonic antigen level was the most common method in China (87.74%, 880/1 003), and the positive rate was 64.5% (568/880). The correct rate of preoperative imaging tests was 40.7% (280/688). The ratio of peritoneal carcinomatosis index (PCI) scores between 0 and 10 was the highest (59.6%, 170/285). Two hundred and sixty-two (27.0%) patients were performed by totally laparoscopic operation in 971 patients. The resection of primary tumor was performed in 588 of the 817 patients (72.0%). In a total of 457 cases, 253 (55.4%) patients were performed cytoreduction which group scored completeness of cytoreduction (CCR) 0. The postoperative hyperthermic intraperitoneal chemotherapy was implemented in 70 of the 334 cases (21.0%). Among 1 003 cases, 562 cases (56.03%) had complete follow-up data and the median overall survival was 15 months. The primary tumor resection and the CCR scores were affected by the PCI scores. The patients underwent primary tumor resection (187/205 vs. 26/80, χ(2)=105.085, P=0.000) and the patients were performed cytoreduction which scored CCR 0 or CCR 1 (162/204 vs. 8/78, Z=-10.465, P=0.000) had significant difference between the groups of PCI<20 and ≥20. There was a close correlation between the surgical method and the CCR scores (Z=-3.246,P=0.001).When the maximum degree of tumor reduction was planned, most surgeons would choose laparotomy. The overall survival time was longer in patients with primary tumor resection (P=0.000). The median survival time was 18.6 months in the group of primary tumor resection. Conclusions: It is difficult to diagnose the synchronous peritoneal carcinomatosis from colorectal cancer before the operation. Primary tumor resection has an obvious effect to prolong the survival time. It is necessary to standardize the treatment of peritoneal metastasis.

The addition of protein-bound amino acids in low-protein diets improves the metabolic and immunological characteristics in fifteen- to thirty-five-kg pigs.

This study was to evaluate the effects of supplementation of AA form (crystalline vs. protein bound) in low-protein diets on growth, metabolic, and immunological characteristics of pigs. A total of 80 barrows (PIC 327 × 1050; 15.57 ± 0.13 kg BW and 48 ± 2 d of age), housed in 4 pigs per pen with 5 pens per treatment, were assigned to 4 dietary treatments of 17, 15, and 13% CP and 13% CP plus casein for 28 d. The crystalline AA were supplemented to meet the requirement of indispensable AA in pigs. Results showed that pigs fed the 13% CP diet or the 13% CP plus casein diet had lower ( < 0.01) ADG and ADFI and a greater ( < 0.01) feed:gain ratio than pigs fed the 17% CP or 15% CP diets over the 4-wk study period. Compared with other diets, pigs fed the 13% CP diet had decreased concentrations of plasma urea nitrogen, albumin ( < 0.01), and mRNA expressions of Toll-like receptor 4 (), nuclear factor kappa B (; < 0.05), and Toll-interacting protein (; < 0.01) in the ileum and also increased activity of plasma glutamate-pyruvate transaminase ( < 0.05) and concentrations of IL-1ß ( < 0.05) and tumor necrosis factor-α ( < 0.01); however, these characteristics were partly normalized by feeding the 13% CP plus casein diet. Furthermore, the plasma concentration of insulin-like growth factor 1 (IGF-1; < 0.01) and mRNA expressions of protein kinase B (), mammalian target of rapamycin (), and ribosomal protein S6 kinase () in longissimus muscle were increased ( < 0.05) in pigs fed the 13% CP plus casein diet relative to pigs fed the 17% CP or 15% CP diets. In summary, reducing dietary CP level from 17% to 15% had no effect on growth, metabolic, and immunological characteristics of 15- to 35-kg pigs. A further reduction of dietary CP level up to 13% would lead to poor growth performance, but metabolic and immunological characteristics were partly normalized using protein-bound AA to replace synthesized AA in the 13% CP diet.

Dietary supplementation with tributyrin prevented weaned pigs from growth retardation and lethal infection via modulation of inflammatory cytokines production, ileal expression, and intestinal acetate fermentation.

Weanling pigs, with an underdeveloped intestine and immature immune system, are usually subjected to depressed feed intake, growth retardation, and postweaning diarrhea. The aim of this study was to determine 1) the growth response of weaned pigs to supplemental tributyrin (TB) and 2) the potential effects and mechanisms of TB in modulating immune responses of lipopolysaccharide (LPS)-challenged piglets. A total of 240 piglets (Duroc × Large White × Landrace) were weaned at 21 d of age to a control (basal diet), supplemented with antibiotics (AB; +AB), supplemented with TB (+TB), or with supplemental AB and TB (+AB+TB) diets, with 10 replicate pens (6 piglets/pen) per diet. At 49 d of age, male pigs from the control and +TB groups were intraperitoneally injected with LPS (25 µg/kg BW) or saline ( = 6) and sacrificed at 4 h after injection to collect blood, intestine, and digesta samples for biochemical analysis. There were higher ( < 0.05) feed intake and lower ( < 0.05) percentage of negative growth piglets in the +TB groups than in the control group during the first week after weaning. For piglets without LPS challenge, there were higher ( < 0.05) ileal fibroblast growth factor 19 () mRNA abundance and total bile acid concentrations in the +TB groups than in the control group, whereas downregulated ( < 0.05) expression was observed in the +TB groups after LPS challenge. Lipopolysaccharide challenge in the control group increased ( < 0.05) plasma tumor necrosis factor α and IL-6 concentrations and colonic amount and decreased ( < 0.05) colonic goblet cells and colonic and cecal acetate concentrations, with no differences ( > 0.05) observed between +TB groups following LPS challenge. Taken together, dietary supplementation with TB prevented growth retardation through stimulating the appetite of weaned pigs and protected piglets against lethal infection via modulation of inflammatory cytokines production, ileal expression, and intestinal acetate fermentation.

Multigene testing of moderate-risk genes: be mindful of the missense.

BACKGROUND: Moderate-risk genes have not been extensively studied, and missense substitutions in them are generally returned to patients as variants of uncertain significance lacking clearly defined risk estimates. The fraction of early-onset breast cancer cases carrying moderate-risk genotypes and quantitative methods for flagging variants for further analysis have not been established. METHODS: We evaluated rare missense substitutions identified from a mutation screen of ATM, CHEK2, MRE11A, RAD50, NBN, RAD51, RINT1, XRCC2 and BARD1 in 1297 cases of early-onset breast cancer and 1121 controls via scores from Align-Grantham Variation Grantham Deviation (GVGD), combined annotation dependent depletion (CADD), multivariate analysis of protein polymorphism (MAPP) and PolyPhen-2. We also evaluated subjects by polygenotype from 18 breast cancer risk SNPs. From these analyses, we estimated the fraction of cases and controls that reach a breast cancer OR≥2.5 threshold. RESULTS: Analysis of mutation screening data from the nine genes revealed that 7.5% of cases and 2.4% of controls were carriers of at least one rare variant with an average OR≥2.5. 2.1% of cases and 1.2% of controls had a polygenotype with an average OR≥2.5. CONCLUSIONS: Among early-onset breast cancer cases, 9.6% had a genotype associated with an increased risk sufficient to affect clinical management recommendations. Over two-thirds of variants conferring this level of risk were rare missense substitutions in moderate-risk genes. Placement in the estimated OR≥2.5 group by at least two of these missense analysis programs should be used to prioritise variants for further study. Panel testing often creates more heat than light; quantitative approaches to variant prioritisation and classification may facilitate more efficient clinical classification of variants.

Effects of Dietary Supplementation of ß-hydroxy-ß-methylbutyrate on Sow Performance and mRNA Expression of Myogenic Markers in Skeletal Muscle of Neonatal Piglets.

The effects of dietary ß-hydroxy-ß-methylbutyrate (HMB) supplementation during gestation on reproductive performance of sows and the mRNA expression of myogenic markers in skeletal muscle of neonatal pigs were determined. At day 35 of gestation, a total of 20 sows (Landrace × Yorkshire, at third parity) were randomly assigned to two groups, with each group receiving either a basal diet or the same diet supplemented with 4 g/day ß-hydroxy-ß-methylbutyrate calcium (HMB-Ca) until parturition. At parturition, the total and live litter size were not markedly different between treatments, however, the sows fed HMB diet had a decreased rate of stillborn piglets compared with the sows fed the control (CON) diets (p < 0.05). In addition, piglets from the sows fed HMB diet tended to have an increased birth weight (p = 0.08), and a reduced rate of low birth weight piglets (p = 0.05) compared with piglets from the CON sows. Nevertheless, lower feed intake during lactation was observed in the sows fed the HMB diet compared with those on the CON diet (p < 0.01). The relative weights of the longissimus dorsi (LD) and semitendinosus (ST) muscle were higher (p < 0.05) in neonatal pigs from the HMB than the CON sows. Furthermore, maternal HMB treatment increased the mRNA levels of the myogenic genes, including muscle regulatory factor-4 (MRF4, p < 0.05), myogenic differentiation factor (MyoD) and insulin-like growth factor-1 (IGF-1, p < 0.01). In conclusion, dietary HMB supplementation to sows at 4 g/day from day 35 of gestation to term significantly improves pregnancy outcomes and increases the expression of myogenic genes in skeletal muscle of neonatal piglets, but reduces feed intake of sows during lactation.

Characterization of FeDREB1 promoter involved in cold- and drought-inducible expression from common buckwheat (Fagopyrum esculentum).

C-repeat-binding factor (CBF)/dehydration-responsive element (DREB) transcription factors play key roles in plant stress responses. However, little information is available on the regulation of CBF/DREB expression. In this study, we isolated and characterized the FeDREB1 promoter sequence from the common buckwheat accession Xinong 9976. To identify the upstream region of the FeDREB1 gene required for promoter activity, we constructed a series of FeDREB1 promoter deletion derivatives. Each deletion construct was analyzed through Agrobacterium-mediated transient transformation in tobacco leaves treated with 4°C cold or drought stress. Promoter-beta-glucuronidase fusion assays revealed that the pCD1 (-270 bp) deletion in the upstream region of FeDREB1 could activate expression of the GUS gene at 4°C. The pCD1 (-270 bp), pCD2 (-530 bp), and pCD3 (-904 bp) deletion induced low-level GUS expression under drought stress. However, the pCD4 (-1278 bp) deletion clearly activated GUS gene expression. Our results suggest that sections pCD1 (-270 bp) and pCD4 (-1278 bp) in the FeDREB1 gene promoter are new sources of induced promoters for adversity-resistance breeding in plant genetic engineering.

Ginkgo biloba extract for essential hypertension: a systemic review.

BACKGROUND: Ginkgo biloba extract (GBE), a traditional natural herbal product, is often used in the treatment of essential hypertension (EH) as complementary therapy in China and European countries. AIM: To critically assess the current clinical evidence of efficacy and safety of GBE for EH. METHODS: 7 electronic databases (Cochrane Library, PubMed, EMBASE, VIP, CBM, Wanfang data, and CNKI) were searched to identify randomized controlled trials (RCTs) of GBE for EH. Methodological quality was assessed independently using the Cochrane Handbook for Systematic Reviews of Interventions. RESULTS: A total of 9 RCTs with 1012 hypertensive patients were identified and reviewed. Most RCTs were of high risk of bias with flawed study design and poor methodological quality. 6 trials demonstrated potential positive effect of GBE as complementary therapy on BP reduction when compared with antihypertensive drug therapy; however, it was not associated with a statistically significant effect on both SBP and DBP reduction in 3 other trials. Despite the positive findings, there were so many methodological limitations and significant clinical heterogeneity. Most of the trials did not report adverse effects, and the safety of GBE is still uncertain. CONCLUSION: No confirmative conclusions on the efficacy and safety of GBE for EH could be drawn. More rigorous trials are warranted to support their clinical use.

Why clinical modulation of efflux transport at the human blood-brain barrier is unlikely: the ITC evidence-based position.

Drug interactions due to efflux transport inhibition at the blood-brain barrier (BBB) have been receiving increasing scrutiny because of the theoretical possibility of adverse central nervous system (CNS) effects identified in preclinical studies. In this review, evidence from pharmacokinetic, pharmacodynamic, imaging, pharmacogenetic, and pharmacovigilance studies, along with drug safety reports, is presented supporting a low probability of modulating transporters at the human BBB by currently marketed drugs.

[Structure determination of the constituents from Citrus grandis Osbeck].

OBJECTIVE: To isolate the chemical constituents of the peelings of Citrus grandis and determine their structure. METHOD: Column chromatography and PTLC were applied to isolating and purifying the chemical constituents, and 1H NMR and 13C NMR were used to elucidate their structure. RESULT: 4 compounds were isolated as meranzin hydrate(1), roseoside(2), 8-(3-beta-D-syl-2-hydrox-3-methylbutyl)-7-methoxycoumarin(3) and glucose(4). CONCLUSION: Compound 2 was isolated as a carotenoid from this genus for the first time.

Huperzine-A in capsules and tablets for treating patients with Alzheimer disease.

AIM: To compare the efficacy and safety between huperzine-A (Hup) in capsules and tablets for treating patients with Alzheimer disease (AD). METHODS: Using multicenter, prospective, double-blind, double-mimic, parallel, positive controlled and randomized methods, 60 patients meeting with the NINCDS-ARDRA criteria of AD were divided into 2 equal groups. Patients in the capsule group received 4 capsules of Hup (each contains 50 micrograms) and 4 tablets of placebo (lactose and starch inside); while the tablet group received 4 tablets of Hup (each contains 50 micrograms) and 4 capsules of placebo, p.o., twice a day for 60 d. All the patients were evaluated with a lot of related ranting scales, and physiological and laboratory examination. RESULTS: There were significant differences (P < 0.01) on all the psychological evaluations between 'before' and 'after' the 60-d trial of 2 groups, but there was no significant difference between 2 groups by group t test (P > 0.05). The changes of oxygen free radicals in 2 groups showed marked improvement. No severe side effect besides moderate to mild nausea was found in both groups. CONCLUSION: There is equal efficacy and safety between Hup in capsule and tablet for treating patients with AD, and Hup can reduce the pathological changes of the oxygen free radicals in the plasma and erythrocytes of patients with AD.

Prevention and therapy of hepatitis B.

PURPOSE: To review the achievements of the prevention and therapy of hepatitis B (HB) in China during the thirty years after the discovery of hepatitis B virus (HBV) in 1967. DATA SOURCE: Data come from an English language literature search using MEDLINE 1966-1999, especially 1982-1999, and Chinese language literature, published and unpublished, from the Chinese Health Administration. STUDY SELECTION: After reviewing of the literature we select 14 articles to address our statement. DATA EXTRACTION: Two observers independently evaluated the quality of the data materials obtaining from these articles. RESULTS: Investigation of HBV vaccine development in China was almost simultaneously with the same kind of work in the international. At least 200 million people have accepted the vaccination of HBV vaccine. The vaccination of the HBV vaccine in China has decreased the prevalence of HBV infection no matter in urban cities or in rural area. The protection against HBV infection can be maintained at least for 9-11 years. CONCLUSIONS: Vaccination of HBV vaccine in China is successful and has obtained great achievements in the prevention and therapy of HB.

Structural alignment of the (+)-trans-anti-benzo[a]pyrene-dG adduct positioned opposite dC at a DNA template-primer junction.

This study reports on the solution conformation of the covalent (+)-trans-anti-[BP]dG adduct (derived from the binding of the highly mutagenic and tumorigenic (+)-anti-benzo[a]pyrene diol epoxide to the N2 of deoxyguanosine) positioned opposite dC at a junctional site in the d(A1-A2-C3-[BP]G4-C5- T6-A7-C8-C9-A10-T11-C12-C13).d(G14-G15-A16-T17-+ ++G18-G19-T20-A21-G22-C23) 13/10-mer DNA sequence. The 13-mer represents the template strand containing the junction [BP]dG4 lesion while the complementary 10-mer models a primer strand which extends upto and is complementary to the modified dG4 residue. The solution conformation has been determined by initially incorporating intramolecular and intermolecular proton-proton distances defined by lower and upper bounds deduced from NOESY spectra as restraints in molecular mechanics computations in torsion angle space and subsequently through restrained molecular dynamics calculations based on a NOE distance and intensity refinement protocol. The duplex segment retains a minimally perturbed B-DNA conformation with all base pairs, including the junctional [BP]dG4.dC23 pair, in Watson-Crick hydrogen-bonded alignments. The pyrenyl ring is not stacked over the adjacent dC5.dG22 base pair but is positioned on the minor groove-side of the [BP]dG moiety and directed toward the 5'-end of the template strand. The pyrenyl ring stacks over the base of the non-adjacent dA2 residue in one direction and the sugar ring of dC23 in the other direction. The solution structure of the (+)-trans-anti-[BP]dG adduct opposite dC in the 13/10-mer in which the modified deoxyguanosine adopts an anti glycosidic torsion angle (this study) is in striking contrast to the structure of the same (+)-trans-anti-[BP]dG moiety in a 13/9-mer of the same sequence but without the dC23 residue positioned opposite the adduct site [Cosman, M., et al. (1995) Biochemistry 34, 15334-15350]. For the latter case, the aromatic portion of the BP residue stacks over the adjacent dC5.dG22 base pair, the modified deoxyguanosine adopts a syn glycosidic torsion angle and is displaced toward the major groove direction. Insights into the factors that affect the sequence and context dependent conformations of stereoisomeric [BP]dG lesions have emerged following comparison of these two structures with the minor groove conformations of the same (+)-trans-anti-[BP]dG lesion in the fully complementary 11-mer duplex [Cosman, M., et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 1914-1918] and in the base displaced-intercalative conformation of the 11/10-mer deletion duplex containing a -1 deletion site opposite the lesion [Cosman, M., et al. (1994) Biochemistry 33, 11507-11517]. The contributing factors where applicable include Watson-Crick base pairing at the site of the lesion, positioning of the carcinogen within the floor of the minor groove, and the tendency of the bulky hydrophobic aromatic BP residue to assume stacked or intercalative conformations.

Major groove (S)-alpha-(N6-adenyl)styrene oxide adducts in an oligodeoxynucleotide containing the human N-ras codon 61 sequence: conformations of the S(61,2) and S(61,3) sequence isomers from 1H NMR.

The (S)-alpha-(N6-adenyl)styrene oxide adducts at positions X6 in d(CGGACXAGAAG). d(CTTCTTGTCCG) and X7 in d(CGGACAXGAAG).d(CTTCTTGTCCG), incorporating codons 60, 61 (underlined), and 62 of the human n-ras protooncogene, were examined by 1H NMR. These were the S(61,2) and S(61,3) adducts. Chemical shift perturbations were in the 3'-direction from the sites of adduction; upfield changes associated with the styrene aromatic ring current were noted for S-SOA6 H2 and H1', T16 N3H, H6, and CH3 resonances in the S(61,2) oligomer. In the S(61,3) oligomer, S-SOA7 H1', T16 H1', C15 N4Ha, and H5 shifted upfield. The styrene aromatic rings flipped rapidly on the NMR time scale; under these conditions the ortho and meta aromatic protons were equivalent. A sequence effect, in which the S(61,2) adduct equilibrated between two conformers, while the S(61,3) adduct exhibited only a single conformation, was observed. Potential energy minimization of the S(61,2) adduct major conformation yielded a structure in which the styrene ring was oriented in the 3'-direction and interacted primarily with the complementary strand. For the S(61,3) adduct, 291 restraints were obtained from NOE data at three mixing times using relaxation matrix analysis. The emergent structures refined to an average rms difference of 1.3 A, determined by pairwise analysis. These were compared to NOE intensity data; the calculated sixth root residual index was 9.2 x 10-2 at 250 ms. In the refined structure, the styrene ring was also oriented in the 3'-direction and interacted with the complementary strand. The minor conformation of the S(61,2) adduct was not identified. These results contrasted with the diastereomeric R(61,2) and R(61,3) adducts, which underwent slow ring flips on the NMR time scale and for which small sequence effects involving the minimum energy conformation of the styrene ring were observed.

[Clinical study on effect of jinshisan on dissolution of extra-/ intrahepatic biliary stones].

In order to evaluate the role and mechanism of Jinshisan (JSS) in bile stone dissolution, 164 cases of extra-/ intrahepatic biliary stone patients were treated by a collaborated group, the therapeutic effect was observed by B-ultrasonography before and after treatment. Results showed: (1) Among the 164 cases, 26 cases (15.85%) were cured clinically; 73 (44.51%) were effective and 65 (39.63%) ineffective. (2) After treatment, in 84 patients of right hepatolith, the stone were diappeared in 10 cases, the size or number of stone decreased in 44 casess. In 34 patients with left hepatolith, the figure were 5 and 13 respectively. In 10 cases of choledocholith they were 10 and 10 respectively. While in 21 cases of multiple gallstone, only one case with stones of both sides disappeared, except 3 ineffective cases, others with stone decreased in size or number or disappeared unilaterally. (3) Gallston diappeared in 7 of 10 patients with size of stone 6 months. (5) Analysis of bile showed that JSS could increase the amount of bile acid, decrease the bilirubin, mucous and bicarbonate radical in bile.

Recognition of specific nucleotide bases and cooperative DNA binding by the trans-acting nitrogen regulatory protein NIT2 of Neurospora crassa.

The NIT2 nitrogen regulatory protein of Neurospora is a DNA binding protein which contains a single Cys2/Cys2 type finger motif followed immediately by a highly basic region. Several different approaches were employed to identify nucleotides which appear to be in contact with NIT2 in the DNA-protein complex. Methylation interference and missing contact analyses with the promoter DNA fragment of the L-amino acid oxidase gene showed that all three purines in both of two GATA core sequences and the single adenine residue in each of the complementary TATC sequences were in intimate contact with NIT2. Modification or loss of the three purine residues located between the two GATA core sequences also significantly reduced NIT2 binding, whereas alteration of purines which flank the binding element showed only minor effects. Chemical modification of all six thymine bases in the two GATA and TATC complement core sequences also strongly affected NIT2 binding. High affinity NIT2 binding sites appear to contain at least two GATA core sequences, with single GATA sequences acting only as weak binding sites. Mobility shift experiments with the DNA fragment upstream of nit-3, the structural gene for nitrate reductase, revealed two DNA-NIT2 protein complexes. In complex I, which is formed first, NIT2 was bound to a pair of GATA sites located at -180. In complex II, the paired GATA sites at -180 plus a single GATA site at -290 were all occupied by NIT2. A DNA fragment containing only the single -290 GATA element bound NIT2 very weakly. The affinity of this single GATA for NIT2 was ten to twenty times greater when it was situated on the same DNA fragment with the distant paired GATA elements than when alone.

[Studies on saponins from the leaves of Panax japonicus var. bipinnatifidus(Seem.)Wu et Feng].

Two new saponins, bipinnatifidusoside F1(XII) and F2(XIII), were isolated along with eleven known saponins from the dried leaves of Panax japonicus var.bipinnatifidus(Seem.)Wu et Feng collected in the Range of Qinling Mountains in China. By spectrometric analysis(FAB-MS,1H and 13CNMR,1H-1H COSY), their structures were elucidated as dammar-25(26)-ene-3 beta, 12 beta, 20(S),24 zeta-tetraol-(20-O-beta-D-glucopyranosyl)-3-O- beta-D-glucopyranosyl(1-2)-beta-D-glucopyranoside (bipinnatifidusodie F1) and dammar-22(23) ene-3 beta, 12 beta, 20(S),24 zeta-tetraol-(20-O-beta-D-glucopyranosyl)-3-O-beta- D-glucopyranosyl(1-2)-beta-D-glucopyranoside (bipinnatifidusoside F2), respectively. The known saponins were identified as ginsenoside F1,F2,F3,Rg2,Re,Rd,Rb1,Rb3, 24(S)-pseudoginsenoside F11, panasenoside and majoroside F1. Compared with bipinnatifidusoside F2, majoroside F3 was corrected as dammar-22(23)-ene-3 beta, 12 beta,20(S),24 zeta-tetraol-(20-O-beta-D- glucopyranosyl)-3-O-beta-D-glucopyranoside.

[Further study on dammarane saponins of leaves of Panax japonicus var. major collected in Quinling Mountains China].

In continuation of our studies on the leaves of Panax japonicus var. major (Burk.) Wu et Feng, two new dammarane saponins are isolated. By means of FAB-MS and 13CNMR, their structures were elucidated as dammar-22(23)-ene-3 beta, 6 alpha, 12 beta, 20(S), 24 section-pentaol- (20-O-beta-D-glucopyranosyl)-6-O-alpha-L-rhamnopyranosyl (1-2)-beta-D- glucopyranoside (majorosids F5) and dammar-23(24)-ene-3 beta, 6 alpha, 12 beta, 20(S), 25-pentaol-(20-O-beta-D- glucopyranosyl)-6-O-alpha-L-rhamnopyranosyl(1-2)-beta-D-glucopyranoside (majoroside F6).