Effects of manganese glycine on eggshell quality, eggshell ultrastructure, and elemental deposition in aged laying hens.

Poor eggshell quality of eggs laid by aged laying hens is the major problem affecting the length of the rearing period in the laying hen industry. Trace elements are required and play vital roles in the eggshell quality of laying hens. Appropriate dose of organic microelements is environmentally friendly and sufficient to satisfy the needs of hens because of their greater bioavailability and lower excretion than inorganic forms. The aim of this experiment was to investigate the effects of manganese (Mn) glycine (MG) on eggshell quality, elemental deposition, and eggshell ultrastructure in aged laying hens. A total of 720 Hy-Line Brown hens 70 weeks old were assigned equally to four groups with six replicates of 30 birds each. The hens were fed basal diets (without Mn supplementation) supplemented with 120 mg/kg of Mn from manganese sulfate monohydrate (MSM), or 40, 80, or 120 mg/kg Mn from MG for 12 weeks. Dietary supplementation with 80 mg/kg Mn from MG resulted in the greatest eggshell strength after 6 weeks of treatment (P = 0.047), and in greater eggshell strength than observed in the MSM control after 12 weeks of treatment (P = 0.025). After 12 weeks of treatment, the eggs of hens in the MG groups showed lower mammillary layer thickness in the blunt end, equator, and acute end than observed in the MSM control group (P < 0.001). With the exception of the blunt ends of eggs from hens in the 120 mg/kg MG group, the eggs of hens in the MG groups, compared with the MSM control group, exhibited a lower mammillary layer ratio, and greater palisade layer ratio and effective layer ratio in the blunt end, equator, and acute end (P < 0.001). Dietary supplementation with 80 mg/kg Mn from MG, compared with the MSM control and 40 and 120 mg/kg MG, resulted in the greatest palisade layer thickness and effective layer thickness, and the lowest mammillary layer thickness in the equator (P < 0.001, P = 0.001, P < 0.001, respectively). Furthermore, supplementation with 80 mg/kg Mn from MG exhibited the greatest ratio of the palisade layer and effective layer, and the lowest mammillary layer ratio in the blunt end and equator (all P < 0.001). The Mn content of eggshells in hens-fed diets supplemented with 80 and 120 mg/kg Mn from MG was greater than that in the MSM control and 40 mg/kg MG groups (P = 0.035). Dietary supplementation with 80 or 120 mg/kg Mn from MG resulted in greater tibia Mn content than observed in the 40 mg/kg MG group (P = 0.019), and greater yolk Mn content than observed in the 40 mg/kg MG and MSM control groups (P = 0.018). In conclusion, dietary supplementation with 80 mg/kg Mn from MG, compared with the MSM control (120 mg/kg Mn), may increase the deposition efficiency of Mn, alter eggshell elemental composition, improve eggshell ultrastructure, and enhance eggshell strength in aged laying hens.

[Molecular epidemiological characterization of hypervirulent carbapenem-resistant Klebsiella pneumoniae in a hospital in Henan Province from 2020 to 2022].

Objective: The study investigated the clinical distribution, antimicrobial resistance and epidemiologic characteristics of hypervirulent Carbapenem-resistant Klebsiella pneumoniae (hv-CRKP) in a hospital in Henan Province to provide a scientific basis for antibiotic use and nosocomial infection prevention and control. Methods: A retrospective analysis of the clinical data from the cases was carried out in this study. Clinical data of patients infected with the CRKP strain isolated from the clinical microbiology laboratory of Henan Provincial Hospital of Traditional Chinese Medicine from January 2020 to December 2022 were retrospectively analyzed. A string test, virulence gene screening, serum killing, and a G. mellonella infection model were used to screen hv-CRKP isolates. The clinical characteristics of hv-CRKP and the drug resistance rate of hv-CRKP to twenty-five antibiotics were analyzed using WHONET 5.6. Carbapenemase phenotypic characterization of the hv-CRKP was performed by colloidal gold immunochromatographic assay, and Carbapenemase genotyping, multi-locus sequence typing (MLST) and capsular serotyping of hv-CRKP isolates were performed by PCR and Sanger sequencing. Results: A total of non-duplicate 264 CRKP clinical isolates were detected in the hospital from 2020 to 2022, and 23 hv-CRKP isolates were detected, so the corresponding detection rate of hv-CRKP was 8.71% (23/264). The hv-CRKP isolates in this study were mainly from the intensive care unit (10/23) and neurosurgery department (8/23), and the main sources of hv-CRKP isolates were sputum (10/23) and bronchoalveolar lavage fluid (6/23). The hv-CRKP isolates in this study were highly resistant to ß-lactam antibiotics, fluoroquinolones and aminoglycosides, and were only susceptible to colistin, tigecycline and ceftazidime/avibactam. The detection rate of the blaKPC-2 among 23 hv-CRKP isolates was 91.30% (21/23) and none of the class B and class D carbapenemases were detected. Results of MLST and capsular serotypes showed that ST11 type hv-CRKP was the dominant strain in the hospital, accounting for 56.52% (13/23), and K64 (9/13) and KL47 (4/13) were the major capsular serotypes. Conclusion: The hv-CRKP isolates from the hospital are mainly from lower respiratory tract specimens from patients admitted to the intensive care department and the drug resistance is relatively severe. The predominant strains with certain polymorphisms are mainly composed of the KPC-2-producing ST11-K64 and ST11-KL47 hv-CRKP isolates in the hospital.

Efficacy evaluation of selenium-enriched yeast in laying hens: effects on performance, egg quality, organ development, and selenium deposition.

The aim of this study was to compare the dynamic changes of egg selenium (Se) deposition and deposition efficiency and to evaluate the efficacy of selenium-enriched yeast (SY) in laying hens over the 84 d feeding period after SY supplementation. A total of one thousand one hundred fifty-two 30-wk-old, Hy-Line Brown hens were randomly assigned to 1 of 6 groups (192 laying hens per group) with 6 replicates of 32 birds each, fed a basal diet (without Se supplementation), basal diet with 0.3 mg/kg of Se from sodium selenite (SS) or basal diets with 0.1, 0.2, 0.3, or 0.4 mg/kg of Se from SY, respectively. The results showed that the Se concentrations in the eggs and breasts from hens fed a SY-supplemented diet were significantly higher than those from hens fed a SS-supplemented diet or a basal diet (P < 0.001). There was a positive linear and quadratic correlation between Se concentrations in the eggs from hens fed a SY-supplemented diet and dietary Se supplementation on days 28, 56, and 84 (r2 = 0.931, 0.932, 0.976, P < 0.001; r2 = 0.946, 0.935, 0.976, P < 0.001), respectively. The Se deposition efficiency in whole eggs from hens fed a basal or SY-supplemented diet weresignificantly higher than those in eggs from hens fed a SS-supplemented diet on days 28, 56, and 84 (P < 0.001), respectively. In addition, there was a positive linear and quadratic correlation between Se concentrations in the eggs from hens fed SY-supplemented diet (r2 = 0.655, 0.779, 0.874, 0.781, P < 0.001; r2 = 0.666, 0.863, 0.944, 0.781, P < 0.001) or SS-supplemented diet (r2 = 0.363, P = 0.002; r2 = 0.440, P = 0.002) and number of feeding days. In conclusion, the organic Se from SY has higher bioavailability and deposition efficiency of Se in whole eggs as compared with inorganic Se from SS. The Se concentrations and Se deposition efficiency in the eggs increased from hens fed a SS- or SY-supplemented diet but decreased from hens fed a basal diet with the extension of the experimental duration. The results indicate that the dietary Se supplementation from SY should be limited to a maximum of 0.1 mg Se/kg complete feed when the eggs and meat produced from hens fed a SY-supplemented diet are used as food for humans directly, whereas up to 0.4 mg/kg organic Se from SY can be used to supplement the diets for laying hens when the products are used as raw materials for producing Se-enriched food.

Patient-derived organoids of non-small cells lung cancer and their application for drug screening.

Patient-derived organoids (PDOs) are emerging as preclinical models with promising values in personalized cancer therapy. The purpose of this study was to establish a living biobank of PDOs from patients with non-small cell lung cancer (NSCLC) and to study the responses of PDOs to drugs. PDOs derived from NSCLC were cultured in vitro, and then treated with natural compounds including chelerythrine chloride, cantharidin, harmine, berberine and betaine with series of concentrations (0.5-30 µM) for drug screening. Phenotypic features and treatment responses of established PDOs were reported. Cell lines (H1299, H460 and H1650) were used for drug screening. We successfully established a living NSCLC organoids biobank of 10 patients, which showed similar pathological features with primary tumors. Nine of the 10 patients showed mutations in EGFR. Natural compounds chelerythrine chloride, cantharidin and harmine showed anticancer activity on PDOs and cell lines. There was no significant difference in the 95% confidence interval (CI) for the IC50 value of chelerythrine chloride between PDOs (1.56-2.88 µM) and cell lines (1.45-3.73 µM, p>0.05). PDOs were sensitive to berberine (95% CI, 0.092-1.55 µM), whereas cell lines showed a resistance (95% CI, 46.57-2275 µM, p<0.0001). PDOs had a higher IC50 value of cantharidin, and a lower IC50 value of harmine than cell lines (p<0.05, 7.50-10.45 µM and 4.27-6.50 µM in PDOs, 3.07-4.44 µM and 4.69-544.99 µM in cell lines, respectively). Both PDOs and cell lines were resistant to betaine. Chelerythrine chloride showed the highest inhibitory effect in both models. Our study established a living biobank of PDOs from NSCLC patients, which might be used for high-throughput drug screening and for promising personalized therapy design.

[Analysis of the factors influencing adjuvant chemotherapy decisions for triple negative breast cancer].

Objective: To analyze adjuvant chemotherapy decisions for triple negative breast cancer (TNBC), and explore the influencing factors in the multidisciplinary treatment (MDT) modality. Methods: A retrospective analysis was performed. The cases with invasive TNBC who underwent surgery and MDT discussion for adjuvant treatment in Ruijin Hospital, from April 2013 to June 2015, were recruited. The patients' clinicopathological characteristics were analyzed and adjuvant treatment suggestions from MDT were obtained. Here the chemotherapy decision alteration was defined as a disagreement in chemotherapy or not, or inconsistence in regimens between the attending doctor and the multidisciplinary team. Results: A total of 194 patients aged ≤70 years old were enrolled in the multidisciplinary discussion, and 187 patients (96.4%) were suggested to receive chemotherapy. When compared the opinions of the attending doctor to suggestions of the multidisciplinary team, we found that the percentage of chemotherapy decision alteration reached 22.7% (39/172), of which 94.9% (37/39) were inconsistence in chemotherapy regimens. There were 119 patients who were recommended to receive epirubicin plus cyclophosphamide (EC) followed by docetaxel (T) or weekly paclitaxel (wP) regimens. Before the announcement of results for the E1199 trial, EC-T accounted for 62.5% (55/88), and EC-wP accounted for 37.5% (33/88) for this group of patients. After that, the proportion of EC-T was decreased to 22.6% (7/31) and proportion of EC-wP increased to 77.4%(24/31) (P<0.001). In addition, a total of 20 patients were suggested to receive platinum based chemotherapy. The proportions were 9.3% in cases with invasive ductal carcinoma, and 33.3% in cases with metaplastic carcinoma, respectively (P=0.016). Conclusions: The adjuvant chemotherapy decision for TNBC patients is altered in 22.7% of the patients after MDT discussion. After the announcement of SABCS E1199 results, more patients are suggested to receive EC followed by weekly paclitaxel. There is a lack of detailed evidence for platinum based adjuvant chemotherapy for TNBC, and more patients with metaplastic carcinoma receive platinum based adjuvant chemotherapy.

Suppressive Effects of Copper Sulfate Accumulation on the Spermatogenesis of Rats.

This study investigated the effect of copper sulfate (CuSO4) in the rat spermatogenesis. Forty male rats, weighing 70-80 g, were randomly divided into four groups: control group (CG, 0 mg/kg BW), low-dose group (LG, 100 mg/kg BW), mid-dose group (MG, 200 mg/kg BW), and high-dose group (HG, 400 mg/kg BW). Rats were administered CuSO4 by gavage for 30 days. A variety of measurements were taken including the testis coefficients, the sperm count, the abnormal malformation rate, testosterone (T), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) concentrations in the serum. In addition, glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities, and malondialdehyde (MDA) concentration in the testis were determined. The results showed that in the CuSO4-treated groups, the testis coefficients, sperm count, T, LH, and FSH concentrations, and GSH-Px and SOD activities decreased, while the abnormal malformation rate and MDA concentration increased, compared with the CG. It indicates that CuSO4 exposure impairs the sperm quality and inhibits secretion of sex hormone and gonadotropin, and testis anti-oxidative function, suppressing the rat spermatogenesis.

[Refractory allergic rhinitis serum levels of vitamin D and vitamin D calcium chewable therapeutic value].

Objective:To survey clinical efficacy of vitamin D calcium chewable tablets in the treatment of adults with refractory allergic rhinitis and investigate the level of 1,25-dihydroxyvitamin D3 in the patients' serum.Method:Sixty cases who were dignosed with adult refractory moderate to severe allergic in our hospital outpatient were admitted in our study from May 2014 to November 2015.Sixty cases of healthy people in our hospital medical center were selected as the control group,serum 1,25-dihydroxyvitamin D3 were detected in all patients and normal controls before the treatment.The control group were treated using fluticasone propionate nasal spray and left cetirizine while the experimental group were added with vitamin D calcium chewable tablets on the basis of the control group. Before treatment and after 2 or 4 weeks of treatment,a visual analog scale of the overall classification of nasal symptoms were used of and clinical symptoms evaluation.Result:Compared with healthy controls,the incidence of serum 1,25-dihydroxy vitamin D3 content is:(17.46±4.94)µg/L and the difference was statistically significant(P<0.0).The symptom scores of the treatment group and control group were significantly decreased before treatment and the difference had statistical significance(P<0.05).After four weeks of treatment,the experimental group of the symptom scores were lower than that of control group.The differences were statistically significant(P<0.05).Conclusion:Serum 1,25 dihydroxyvitamin D3 deficiency is a widespread phenonmenon in refractory allergic rhinitis.Vitamin D calcium chewable tables adjuvant treatment of allergic rhinitis is one of methods with safe,effective,and simple treatment.

Effects of RuPeng15 Powder (RPP15) on Monosodium Urate Crystal-Induced Gouty Arthritis in Rats.

RuPeng15 Powder (RPP15) is a herbal multicompound remedy that originates from traditional Tibetan medicine and possesses antigout, anti-inflammatory, and antihyperuricemic properties based on the traditional conceptions. The present study was undertaken to evaluate the therapeutic effect of PRP15 in rat gouty arthritis induced by monosodium urate (MSU) crystals. In the present study, we found that treatment with RPP15 (0.4, 0.8, and 1.2 g/kg) in rats with gouty arthritis induced by MSU crystals significantly attenuated the knee swelling. Histomorphometric and immunohistochemistry analyses revealed that MSU-induced inflammatory cell infiltration and the elevated expressions of nuclear transcription factor-κB p65 (NF-κB p65) in synovial tissues were significantly inhibited, and enzyme-linked immunosorbent assay (ELISA) result showed that MSU-induced high levels of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1ß), and interleukin-8 (IL-8) in synovial fluid were reduced by treatment with RPP15 (0.4, 0.8, and 1.2 g/kg). We conclude that RPP15 may be a promising candidate for the development of a new treatment for gout and its activity of antigout may be partially related to inhibiting TNF-α, IL-1ß, IL-8, and NF-κB p65 expression in the synovial tissues.

Effects of sub-chronic aluminum chloride on spermatogenesis and testicular enzymatic activity in male rats.

AIMS: The aim of this study was to determine the effects of sub-chronic aluminum chloride (AlCl3) on spermatogenesis and testicular enzymatic activity in male rats. MAIN METHODS: Forty Wistar male rats were randomly divided into four groups: control group (CG, 0), low-dose group (LG, 64.18 mg/kg BW AlCl3), mid-dose group (MG, 128.36 mg/kg BW AlCl3) and high-dose group (HG, 256.72 mg/kg BW AlCl3). The rats were orally administered with AlCl3 for 120 days. At the end of the experiment, the contents of Al, Fe, Cu and Zn, the enzyme activities of testicular acid phosphatase (ACP), succinate dehydrogenase (SDH), lactate dehydrogenase (LDH), lactate dehydrogenase isoenzyme (LDH-x), the sperm count and the sperm malformation rate were examined. KEY FINDINGS: The results showed that the Al and Cu contents, sperm count and the enzyme activities of testicular ACP, SDH, LDH and LDH-x decreased, while the Zn and Fe contents and sperm malformation rate increased in AlCl3-treated rats. SIGNIFICANCE: It suggests that sub-chronic AlCl3 disorders the balance of trace element and decreases the spermatogenesis and the activities of testicular enzymes, indicating that AlCl3 has adverse effect on the testicular function in male rats.

Effects of sub-chronic aluminum chloride exposure on rat ovaries.

AIMS: This experiment investigated the effects of sub-chronic aluminum chloride (AlCl3) exposure on rat ovaries. MAIN METHODS: Eighty female Wistar (5weeks old) rats, weighed 110-120g, were randomly divided into four treatment groups: control group (CG), low-dose group (LG, 64mg/kg BW AlCl3), mid-dose group (MG, 128mg/kg BW AlCl3) and high-dose group (HG, 256mg/kg BW AlCl3). The AlCl3 was administered in drinking water for 120days. The ovarian ultrastructure was observed. The activities of acid phosphatase (ACP), alkaline phosphatase (ALP), succinate dehydrogenase (SDH), Na(+)-K(+)-ATPase, Mg(2+)-ATPase and Ca(2+)-ATPase, the contents of Fe, Cu and Zn, and the protein expression of follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) in the ovary were determined. KEY FINDINGS: The results showed that the structure of the ovary was disrupted, the activities of ALP, ACP, SDH, Na(+)-K(+)-ATPase, Mg(2+)-ATPase and Ca(2+)-ATPase, the contents of Zn, Fe and the protein expression of FSHR and LHR were lowered, and the content of Cu was increased in AlCl3-treated rats than those in control. SIGNIFICANCE: The results indicate that sub-chronic AlCl3 exposure caused the damage of the ovarian structure, the disturbed metabolism of Fe, Zn and Cu and the decreased activities of Na(+)-K(+)-ATPase, Mg(2+)-ATPase and Ca(2+)-ATPase in the ovary, which could result in suppressed energy supply in the ovary. A combination of suppression of energy supply and reduction of expression of FSHR and LHR could inhibit ovulation and corpus luteum development, leading to infertility in female rats.

Evidence for inbreeding depression in the tree Robinia pseudoacacia L. (Fabaceae).

The magnitude of inbreeding depression within populations is important for the evolution and maintenance of mixed mating systems. However, data are sparse on the magnitude of inbreeding depression in Robinia pseudoacacia. In this study, we compared differences in the mature seed set per fruit, seed mass, germination success, and seedling growth between self- and cross-pollination treatments and estimated the inbreeding depression at 3 stages: seed maturation, seedling emergence, and seedling growth at 10 and 20 weeks. We found that progenies resulting from cross-pollination treatments showed significantly higher fitness than progenies resulting from self-pollination, causing high levels of inbreeding depression. Inbreeding depression was not uniformly manifested, however, over the 3 stages. Inbreeding depression was the greatest between fertilization and seed maturation stage (δ = 0.5419), and the seedling emergence (0.3654) stage was second. No significant differences in seedling growth were observed between selfed and crossed progenies. The cumulative inbreeding depression (δ) across all 3 stages averaged 0.7452. Inbreeding depression may promote outcrossing in R. pseudoacacia by acting as a post-pollination barrier to selfing. The large difference in the seed set between self- and cross-pollination that we detected indicated that inbreeding depression would probably be a reasonable explanation for the high abortion and low seed set in R. pseudoacacia.

Investigation of the mutagenic potential of cold atmospheric plasma at bactericidal dosages.

In the past few years, cold atmospheric plasma (CAP) has evolved into a new tool in the fight against nosocomial infections and antibiotic-resistant microorganisms. The products generated by the plasma-electrons, ions, reactive species and UV light-represent a 'lethal cocktail' for different kinds of pathogen, which opens up possible applications in hygiene and medicine. Nevertheless, to ensure the safe usage of CAP on skin (e.g., to treat wounds or skin diseases) several pre-clinical in vitro studies have to be performed before implementing clinical trials on humans. In the study presented here, inactivation experiments with Escherichia coli were carried out to identify the necessary plasma dosage for a 5 log reduction: with a small hand-held battery-operated CAP device, these disinfection properties were achieved after application during 30s. This and higher plasma dosages were then used to analyze the mutagenicity induced in V79 Chinese hamster cells-to furthermore define a 'safe application window'-with the HPRT (hypoxanthine-guanine phosphoribosyl transferase) mutation assay. The results show that a CAP treatment of up to 240 s and repeated treatments of 30s every 12h did not induce mutagenicity at the Hprt locus beyond naturally occurring spontaneous mutations.

Promotion of peri-implant bone healing by systemically administered parathyroid hormone (1-34) and zoledronic acid adsorbed onto the implant surface.

UNLABELLED: The effect of human parathyroid hormone 1-34 (PTH) and zoledronic acid (ZA) alone or in combination on bone healing in osteoporotic settings was tested using implants inserted in tibiae of ovariectomized (OVX) rats. Combination therapy promoted bone healing more than each treatment alone 12 weeks after implant insertion. INTRODUCTION: PTH and ZA have been demonstrated to be effective on implant fixation. However, reports about the combined use of PTH and ZA for promotion of bone healing around implant in osteoporotic settings are still limited. This study aims to investigate effects of PTH+ZA on implant stabilization in OVX rats. METHODS: Twelve weeks after bilateral ovariectomy, OVX rats randomly received implants without or with ZA (by immersion in 1 mg/ml ZA solution for 24 h). Subsequently, half of the animals from each group also received subcutaneous injections of PTH (60 µg/kg, three times a week) for 12 weeks. Thus, there were four groups: control, PTH, ZA, and PTH+ZA. RESULTS: All treatments promoted bone healing around implant compared to control, but PTH+ZA treatment showed significantly stronger effects than PTH or ZA alone in histological, micro-CT, and biomechanical tests. CONCLUSION: The results indicated the additive effects of PTH and ZA on implant fixation in OVX rats; it was suggested that the anabolic effect of PTH was potent and not blunted by ZA during bone healing around implant when used concurrently.

Bioactivities of chicken essence.

The special flavor and health effects of chicken essence are being widely accepted by people. Scientific researches are revealing its truth as a tonic food in traditional health preservation. Chicken essence has been found to possess many bioactivities including relief of stress and fatigue, amelioration of anxiety, promotion of metabolisms and post-partum lactation, improvement on hyperglycemia and hypertension, enhancement of immune, and so on. These activities of chicken essence are suggested to be related with its active components, including proteins, dipeptides (such as carnosine and anserine), polypeptides, minerals, trace elements, and multiple amino acids, and so on. Underlying mechanisms responsible for the bioactivities of chicken essence are mainly related with anti-stress, anti-oxidant, and neural regulation effects. However, the mechanisms are complicated and may be mediated via the combined actions of many active components, more than the action of 1 or 2 components alone.

Assessment of genetic diversity and variation of Robinia pseudoacacia seeds induced by short-term spaceflight based on two molecular marker systems and morphological traits.

The black locust (Robinia pseudoacacia) is a forest legume that is highly valued as a honey plant and for its wood. We explored the effect of short-term spaceflight on development of R. pseudoacacia seedlings derived from seeds that endured a 15-day flight; the genetic diversity and variation of plants sampled from space-mutagenized seeds were compared to plants from parallel ground-based control seeds using molecular markers and morphological traits. In the morphology analysis, the space-mutagenized group had apparent variation compared with the control group in morphological traits, including plant height, basal diameter, number of branches, branch stipular thorn length, branch stipular thorn middle width, leaflet vertex angle, and tippy leaf vertex angle. Simple sequence repeat (SSR) and sequence-related amplified polymorphism (SRAP) molecular marker analyses showed a slightly higher levels of genetic diversity in the space-mutagenized group compared to the control group. In the SRAP analysis, the space-mutagenized group had 115 polymorphic bands vs 98 in the controls; 91.27% polymorphic loci vs 77.78% in the controls; 1.9127 ± 0.2834 alleles vs 1.7778 ± 0.4174 in the controls; Nei's genetic diversity (h) was 0.2930 ± 0.1631 vs 0.2688 ± 0.1862 in the controls, and the Shannon's information index (I) was 0.4452 ± 0.2177 vs 0.4031 ± 0.2596 in the controls. The number of alleles was significantly higher in the space-mutagenized group. In the SSR analysis, the space-mutagenized group also had more polymorphic bands (51 vs 46), a greater percentage of polymorphic loci (89.47% vs 80.70%); h was also higher (0.2534 ± 0.1533 vs 0.2240 ± 0.1743), as was I (0.3980 ± 0.2069 vs 0.3501 ± 0.2412). These results demonstrated that the range of genetic variation in the populations of R. pseudoacacia increased after spaceflight. It also suggested that the SSR and SRAP markers are effective markers for studying mutations and genetic diversity in R. pseudoacacia. The data provide valuable molecular evidence for the effects of the space environment on R. pseudoacacia and may contribute to future space-breeding programs involving forest trees.

The effects of combined human parathyroid hormone (1-34) and zoledronic acid treatment on fracture healing in osteoporotic rats.

UNLABELLED: Ovariectomized (OVX) rats with tibial fracture received vehicle, ZA, PTH, or ZA plus PTH treatment for 4 and 8 weeks. Bone metabolism, callus formation, and the mass of undisturbed bone tissue were evaluated by serum analysis, histology, immunohistochemistry, radiography, micro-computerized tomography, and biomechanical test. INTRODUCTION: Previous studies have demonstrated the effect of ZA or PTH on osteoporotic fracture healing. However, reports about effects of ZA plus PTH on callus formation of osteoporotic fracture were limited. This study was designed to investigate the impact of combined treatment with ZA and PTH on fracture healing in OVX rats. METHODS: Twelve weeks after bilateral ovariectomy, all rats underwent unilateral transverse osteotomy on tibiae. Animals then randomly received vehicle, ZA (1.5 µg/kg weekly), PTH (60 µg/kg, three times a week), or ZA plus PTH until death at 4 and 8 weeks. The blood and bilateral tibiae of rats were harvested for evaluation. RESULTS: All treatments increased callus formation and strength other than the control; ZA + PTH showed the strongest effects on percent bone volume (BV/TV), trabecular thickness, total fluorescence-marked callus area, and biomechanical strength. Additionally, inhibited RANKL and enhanced osteoprotegerin expression were observed in the ZA + PTH group. But no difference in bone mineral density and BV/TV of the contralateral tibiae was observed between treated groups. CONCLUSION: Findings in this study suggested an additive effect of ZA and PTH on fracture healing in OVX rats, and this additive effect was specific to callus formation, not to undisturbed bone tissue.

Systemic treatment with strontium ranelate promotes tibial fracture healing in ovariectomized rats.

UNLABELLED: Systemic treatment with strontium ranelate (SR) was performed on ovariectomized (OVX) rats with fractured tibiae. Callus quality was assessed by radiographic, histological, micro-computerized tomography, and biomechanical examinations at 4 and 8 weeks after fracture. Results revealed that systemic applied SR promoted osteoporotic fracture healing. INTRODUCTION: Several studies have demonstrated the dual effect of SR on osteoporotic and undisturbed bone. However, reports of their effect on osteoporotic fracture healing are limited. This study was designed to investigate the effects of SR on bone regeneration in OVX rats with fractured tibiae. METHODS: Three months after being OVX, female Sprague-Dawley rats accepted bilateral osteotomy on proximal tibiae fixed with intramedullary wires and were divided into two groups: OVX and OVX + SR (625 mg/kg/day). Callus quality was evaluated at 4 and 8 weeks postfracture. RESULTS: Compared with OVX group, SR treatment significantly increased bone formation, BMD, biomechanical strength, and improved microstructural properties of the callus. The ultimate load was increased by 211.0% and 61.4% (p<0.01), and the total bone volume of callus by 74.8% and 79.3% (p<0.01) at 4 and 8 weeks postfracture, respectively. SR treatment also promoted healing progress with increased osteogenesis at 4 weeks; more mature and tightly arranged woven or lamellar bone at 8 weeks across the fracture gap in histological analysis. CONCLUSION: This study suggests that systemic treatment with strontium ranelate could promote tibial fracture healing in OVX rats.

Synthesis and biological activity of Wuweizisu C and analogs.

Lignans are widely distributed in nature. The earliest recorded medicinal use of lignans dated back to over 1000 years ago. Lignan-rich plant products were also active ingredients in Chinese and Japanese folk medicines for the treatment of various diseases. The dried root and stem of this plant are listed in the Chinese pharmacopoeia for the treatment of rheumatoid arthritis, gastric, duodenal ulcers and many other diseases. This review highlights synthetic strategies for the Wuweizisu C analogs and the important pharmacological activities as well as therapeutic findings related to the treatment of HBV and other diseases. Notably a significant and ongoing project on Wuweizisu C and its analogs has led to the discovery and development of two potent derivatives alpha-DDB and BICYCLOL which are currently in clinical trials against HBV, especially in lowering elevated SGPT levels. Further design, synthesis, and evaluation of Wuweizisu C analogs are discussed.

Effects of overexpression of endogenous phenylalanine ammonia-lyase (PALrs1) on accumulation of salidroside in Rhodiola sachalinensis.

Salidroside, a novel effective adaptogenic drug extracted from the medicinal plant Rhodiola sachalinensis A. Bor, can be derived from phenylalanine or tyrosine. Due to the scarcity of R. sachalinensis and its low yield of salidroside, there is great interest in enhancing production of salidroside by the plant. In this study, a cDNA clone encoding phenylalanine ammonia-lyase (PAL) was isolated from R. sachalinensis using rapid amplification of cDNA ends. The resulting cDNA was designated PALrs1. It is 2407-bp long and encodes 710 deduced amino acid residues. Southern blot analysis of genomic DNA indicated that the PAL gene family is composed of three to five genes in the R. sachalinensis genome. Northern blot analysis revealed that transcripts of PALrs1 were present in calli, leaves and stems, but expression in roots was very low. The PALrs1 under the 35S promoter with double-enhancer sequences from CaMV-Omega and TMV-Omega fragments was transferred into R. sachalinensis via Agrobacterium tumefaciens. PCR and PCR-Southern blot confirmed that the PALrs1 gene had been integrated into the genome of transgenic plants. Northern blot analysis revealed that the PALrs1 gene had been expressed at the transcriptional level. High-performance liquid chromatography indicated that overexpression of the PALrs1 gene resulted in a 3.3-fold increase in p-coumaric acid content, as expected. In contrast, levels of tyrosol and salidroside were 4.7-fold and 7.7-fold, respectively, lower in PALrs1 transgenic plants than in controls. Furthermore, overexpression of the PALrs1 gene resulted in a 2.6-fold decrease in tyrosine content. These data suggest that overexpression of the PALrs1 gene and accumulation of p-coumaric acid did not facilitate tyrosol biosynthesis; tyrosol, as a phenylethanoid derivative, is not derived from phenylalanine; and reduced availability of tyrosine most likely resulted in a large reduction in tyrosol biosynthesis and accumulation of salidroside.

Japonicins A and B from the flowers of Inula japonica.

Two new flavonols japonicins, A (1) and B (2), along with nine known flavonoids were isolated from an antidepressant fraction, which was separated from the 70% alcohol extract of the flowers of Inula japonica Thumb. The structures of compounds 1 and 2 were determined as 3,3',4',5,9,10-hexahydroxy-12-methylchroman[2,3-h]flavone and 8-(1-(3,4-dihydroxyphenyl)ethyl)-3,3',4',5,7-pentahydroxyflavone by the analyses of physical constants and spectral data. The special flavonoid structure having the substituent 1-phenylethyl at C-8 position was discovered for the first time.