Potent immunosuppressive and anti-inflammatory bisindole alkaloids from Melodinus fusiformis.

Phytochemical investigation of Melodinus fusiformis led to a new aspidosperma-aspidosperma bisindole alkaloid (BIA), bis-19ß-hydroxyvenalstonidine (1), together with three known BIAs (2-4). The structures were established by extensive analysis of their HRESIMS, NMR data, and comparing with the reported data. BIA 1 is an almost symmetrical structure, linked by C3-C14' bond, while BIAs 2-4 are reported for the first time from the plant. The cytotoxic, immunosuppressive and anti-inflammatory activities of BIAs 1-4 were evaluated in vitro. BIAs 1, 3 and 4 showed good toxicity against MOLT-4 cell lines with IC50 values in the range of 1.5-17.5 -M. BIA 2 exhibited the strongest inhibitory effect against MCF-7 cell lines with an IC50 value of 7.1 µM. BIA 1 significantly inhibited Con A-stimulated mice splenocytes proliferation equal to that of the positive control (DXM) in a concentration-dependent manner. BIAs 1 and 2 were able to decrease the NO production in LPS-induced RAW 264.7 cells at 30 µM concentration. BIA 2 showed similar inhibition of nitric oxide release, compared to that of DXM. Furthermore, BIA 2 remarkably inhibited the levels of IL-6 and TNF-α compared to the LPS induced group. Interestingly, BIA 2 displayed an inhibitory effect on TNF-α production similar to that of dexamethasone at a concentration of 20 µM.

HRESIMS-guided isolation of aspidosperma-scandine type bisindole alkaloids from Melodinus cochinchinensis and their anti-inflammatory and cytotoxic activities.

The Melodinus species have been proved to be good resources of bisindole alkaloids. Six bisindole alkaloids were isolated from the leaves and stems of Melodinus cochinchinensis (Lour.) Merr. guided by HRESIMS data analysis. Among them, melokhanines K-M, epi-scandomelonine, and epi-scandomeline possessed aspidosperma-scandine skeleton linked by a C-C bond while meloyine II had a scandine-scandine skeleton. The structures were established by extensive spectroscopic analysis of their HRESIMS and NMR data. Melokhanines K-M were undescribed compounds, while epi-scandomelonine, epi-scandomeline and meloyine II were known compounds, which were reported from Melodinus species for the first time. The anti-inflammatory and cytotoxic activities of the isolates were also evaluated in vitro. Melokhanine K and meloyine II showed potent inhibitory activity on the production of nitric oxide, interleukin-6, and tumor necrosis factor-α in LPS-induced RAW 264.7 macrophages, whereas epi-scandomelonine and epi-scandomeline exhibited certain cytotoxic activity against MOLT-4 cells with IC50 values 5.2 and 1.5 µM, respectively.

Clinical Characteristics of Children with Coronavirus Disease 2019 in Hubei, China.

Since December 2019, COVID-19 has occurred unexpectedly and emerged as a health problem worldwide. Despite the rapidly increasing number of cases in subsequent weeks, the clinical characteristics of pediatric cases are rarely described. A cross-sectional multicenter study was carried out in 10 hospitals across Hubei province. A total of 25 confirmed pediatric cases of COVID-19 were collected. The demographic data, epidemiological history, underlying diseases, clinical manifestations, laboratory and radiological data, treatments, and outcomes were analyzed. Of 25 hospitalized patients with COVID-19, the boy to girl ratio was 1.27:1. The median age was 3 years. COVID-19 cases in children aged <3 years, 3.6 years, and ≥6-years patients were 10 (40%), 6 (24%), and 9 (36%), respectively. The most common symptoms at onset of illness were fever (13 [52%]), and dry cough (11 [44%]). Chest CT images showed essential normal in 8 cases (33.3%), unilateral involvement of lungs in 5 cases (20.8%), and bilateral involvement in 11 cases (45.8%). Clinical diagnoses included upper respiratory tract infection (n=8), mild pneumonia (n=15), and critical cases (n=2). Two critical cases (8%) were given invasive mechanical ventilation, corticosteroids, and immunoglobulin. The symptoms in 24 (96%) of 25 patients were alleviated and one patient had been discharged. It was concluded that children were susceptible to COVID-19 like adults, while the clinical presentations and outcomes were more favorable in children. However, children less than 3 years old accounted for majority cases and critical cases lied in this age group, which demanded extra attentions during home caring and hospitalization treatment.

Evans-Showell-type polyoxometalate-based metal-organic complexes with novel 3D structures constructed from flexible bis-pyrazine-bis-amide ligands and copper metals: syntheses, structures, and fluorescence and catalytic properties.

Two new Evans-Showell-type polyoxometalate (POM)-based metal-organic complexes, namely {Cu3(L1)1.5(H2O)5[Co2Mo10H4O38]}·5H2O (1), {[Cu(L2)0.5(H2O)2]2[Co2Mo10H4O38]}·6H2O (2) (L1 = N,N'-bis(2-pyrazinecarboxamide)-1,4-butane, L2 = N,N'-bis(2-pyrazinecarboxamide)-1,6-hexane), were successfully synthesized and structurally characterized by single-crystal X-ray diffraction, elemental analysis, IR spectroscopy, powder X-ray diffraction (PXRD) and thermogravimetric analyses (TGA). In complex 1, the adjacent [Co2Mo10H4O38]6- polyoxoanions are linked by CuII ions to form a 1D Cu-[Co2Mo10H4O38]6- inorganic chain, which is further linked by ligand L1 and [Co2Mo10H4O38]6- polyoxoanions, forming a 3D metal-organic framework. In complex 2, the adjacent [Co2Mo10H4O38]6- polyoxoanions link the CuII ions to generate a 2D Cu-[Co2Mo10H4O38]6- inorganic layer, which is further connected with bidentate ligands L2 to obtain a 3D metal-organic framework. The structural diversities of compounds 1 and 2 showed that the spacer lengths of the flexible bis-pyrazine-bis-amide ligands play important roles in tuning the structures of the title complexes. Compounds 1 and 2 represent the first examples of 3D frameworks based on the Evans-Showell-type polyoxoanions and Cu-bis-pyrazine-bis-amide coordination complexes. Moreover, the ligand L1 was first successfully introduced into the POM system. The electrochemical and fluorescence properties of compounds 1 and 2 were discussed. As heterogeneous catalysts, compounds 1 and 2 have good catalytic activity for the oxidation of benzyl alcohol. Moreover, compound 2 has higher catalytic performance with 100% conversion and 98.0% selectivity for benzoic acid at 10 h. The difference in their catalytic performance may be mainly due to the difference of their structures. The catalysts can be recovered and reused without displaying any significant loss of activity.

[Effects of thinning intensity on Pinus tabulaeformis plantation in Huanglong Mountain, Northwest China: a comprehensive evaluation].

A sampling plot investigation was conducted on the seedling regeneration, stand growth, species diversity, and soil characteristics in a Pinus tabulaeformis plantation in Huanglong Mountain on the Loess Plateau of Northwest China after 7 years of close-to-natural management thinning 15% (light thinning), 30% (medium thinning), and 45% (heavy thinning), with the effect of different thinning intensities evaluated. With the increase of thinning intensity, the amount and growth indices of 1-7 years old P. tabulaeformis seedlings increased, but the mean annual increments of the growth indices decreased after an initial increase, with the maximum under medium thinning. As compared with the control (un-thinned plantation), the individual volume under light, medium, and heavy thinning was increased by 20.9%, 32.1% and 52.6%, the volume per hm2 decreased by 4.4%, 15.1%, and 25.3%, and the mean annual growth rate of volume increased by 28.6%, 46.2% and 82.0%, respectively. The species diversity and soil characteristics were improved under thinning, with the order of heavy thinning > medium thinning > light thinning > un-thinning. In this study, 45% thinning was most suitable to the management of P. tabulaeformis plantation in Huanglong Mountain.

Genome-wide association study identifies ALDH7A1 as a novel susceptibility gene for osteoporosis.

Osteoporosis is a major public health problem. It is mainly characterized by low bone mineral density (BMD) and/or low-trauma osteoporotic fractures (OF), both of which have strong genetic determination. The specific genes influencing these phenotypic traits, however, are largely unknown. Using the Affymetrix 500K array set, we performed a case-control genome-wide association study (GWAS) in 700 elderly Chinese Han subjects (350 with hip OF and 350 healthy matched controls). A follow-up replication study was conducted to validate our major GWAS findings in an independent Chinese sample containing 390 cases with hip OF and 516 controls. We found that a SNP, rs13182402 within the ALDH7A1 gene on chromosome 5q31, was strongly associated with OF with evidence combined GWAS and replication studies (P = 2.08x10(-9), odds ratio = 2.25). In order to explore the target risk factors and potential mechanism underlying hip OF risk, we further examined this candidate SNP's relevance to hip BMD both in Chinese and Caucasian populations involving 9,962 additional subjects. This SNP was confirmed as consistently associated with hip BMD even across ethnic boundaries, in both Chinese and Caucasians (combined P = 6.39x10(-6)), further attesting to its potential effect on osteoporosis. ALDH7A1 degrades and detoxifies acetaldehyde, which inhibits osteoblast proliferation and results in decreased bone formation. Our findings may provide new insights into the pathogenesis of osteoporosis.

4,5-Diaza-9H-fluoren-9-imine.

In the title compound, C(11)H(7)N(3), the diaza-fluorene rings are almost coplanar with an r.m.s. deviation of 0.0160 Å. In the crystal structure, C-H⋯N hydrogen bonds link mol-ecules into sheets parallel to the ab plane. Mol-ecules are also stacked regularly along the c axis by a variety of π-π inter-actions with centroid-centroid distances in the range 3.527 (2)-3.908 (2) Å.

N-(4,5-Diaza-9H-fluoren-9-yl-idene)-4-meth-oxy-aniline.

In the title compound, C(18)H(13)N(3)O, the diaza-fluorene ring system is almost coplanar (r.m.s. deviation = 0.0640 Å) and subtends an angle of 61.5 (4)° with the plane of the meth-oxy-substituted benzene ring. In the crystal structure, pairs of C-H⋯O hydrogen bonds link mol-ecules into centrosymmetric dimers parallel to the ab plane. Mol-ecules are also stacked in an obverse fashion along the c axis by a variety of π-π inter-actions with centroid-centroid distances in the range 3.557 (2)-3.921 (2) Å.

2,2'-Hexamethyl-enedi-1,3-benzothia-zole.

The title compound, C(20)H(20)N(2)S(2), was prepared by the reaction of suberic acid and 2-amino-thio-phenol under microwave irradiation. The mol-ecule lies on an inversion center.

2-Nitro-N-propyl-benzamide.

The title compound, C(10)H(12)N(2)O(3), contains three mol-ecules in the asymmetric unit. In the crystal structure, inter-molecular N-H⋯O inter-actions link the mol-ecules into chains along the b axis. The crystal structure is consolidated by weak C-H⋯π inter-actions.

Genome-wide copy-number-variation study identified a susceptibility gene, UGT2B17, for osteoporosis.

Osteoporosis, a highly heritable disease, is characterized mainly by low bone-mineral density (BMD), poor bone geometry, and/or osteoporotic fractures (OF). Copy-number variation (CNV) has been shown to be associated with complex human diseases. The contribution of CNV to osteoporosis has not been determined yet. We conducted case-control genome-wide CNV analyses, using the Affymetrix 500K Array Set, in 700 elderly Chinese individuals comprising 350 cases with homogeneous hip OF and 350 matched controls. We constructed a genomic map containing 727 CNV regions in Chinese individuals. We found that CNV 4q13.2 was strongly associated with OF (p = 2.0 x 10(-4), Bonferroni-corrected p = 0.02, odds ratio = 1.73). Validation experiments using PCR and electrophoresis, as well as real-time PCR, further identified a deletion variant of UGT2B17 in CNV 4q13.2. Importantly, the association between CNV of UGT2B17 and OF was successfully replicated in an independent Chinese sample containing 399 cases with hip OF and 400 controls. We further examined this CNV's relevance to major risk factors for OF (i.e., hip BMD and femoral-neck bone geometry) in both Chinese (689 subjects) and white (1000 subjects) samples and found consistently significant results (p = 5.0 x 10(-4) -0.021). Because UGT2B17 encodes an enzyme catabolizing steroid hormones, we measured the concentrations of serum testosterone and estradiol for 236 young Chinese males and assessed their UGT2B17 copy number. Subjects without UGT2B17 had significantly higher concentrations of testosterone and estradiol. Our findings suggest the important contribution of CNV of UGT2B17 to the pathogenesis of osteoporosis.

Polymorphisms in the estrogen receptor genes are associated with hip fractures in Chinese.

INTRODUCTION: Hip fractures (HF) are a major cause of public health burden with strong genetic determination. However, the true causal genes remain largely unknown. MATERIALS AND METHODS: Based on the important biological role of estrogens in bone homeostasis, this study aimed to investigate whether the estrogen receptor genes, ESR1 and ESR2, affect the onset of HF in 700 elderly Chinese subjects (350 with osteoporotic HF and 350 healthy controls). We genotyped 32 SNPs in total and examined their associations both by the single-SNP and haplotype tests. RESULTS: We identified two novel SNPs of ESR1, rs3020314 and rs1884051, were significantly associated with HF (rs3020314: P=0.0004, OR=1.66, 95%CI: 1.25-2.18; rs1884051: P=0.0004, OR=1.46, 95%CI: 1.19-1.81). We firstly detected significant association of ESR2 with HF (rs960070: P=0.0070, OR=1.43, 95%CI: 1.10-1.86). Haplotype analyses corroborated our single-SNP results. CONCLUSION: Our findings have important implications for understanding the pathology of osteoporotic fractures. Independent replication studies are needed to validate our results and explore the most possible functional variants for molecular studies.

[Indexes of intervertebral disc degeneration in rats during the aging process].

OBJECTIVE: To study the indexes for evaluating intervertebral disc degeneration in rats during the aging process. METHODS: Nine SD rats were fed for 6 months and 12 for 22 months as the young and aged groups, respectively. The Miyamoto's grade of the rats was calculated, and the quantity and relative area of the vascular buds as well as the thickness of the calcified and non-calcified layers of the cartilage endplate were measured using the stereoscopic method. Immunohistochemistry with monoclonal antibodies was used to determine the expressions of collagens II and X in the endplate. RESULTS: The quantity and relative area of the vascular buds, non-calcified layer/calcified layer ratio, type II collagen expression in the calcified layer and nucleus pulposus of the cartilage endplate were all significantly decreased in the aged rats as compared with those of the youth rats (P<0.05), but the collagen X expression in the non-calcified layer was significantly higher in the aged rats (P=0.003). No significant difference was found in the Miyamoto's grade between the aged and young rats (P=1.130). CONCLUSION: The relative area of the vascular buds, non-calcified layer/calcified layer ratio, phenotypic expressions of collagens II and X in the cartilage endplate, but not the Miyamoto's grade, are sensitive indexes for evaluating intervertebral disc degeneration in rats during the aging process.

5-(2-Bromo-phen-yl)-1,3,4-thia-diazol-2-amine.

In the title compound, C(8)H(6)BrN(3)S, the thia-diazole ring is oriented at a dihedral angle of 48.35 (3)° with respect to the bromo-phenyl ring. In the crystal structure, inter-molecular N-H⋯N hydrogen bonds link the mol-ecules.

2-[1-Chloro-3-(2-methyl-5-nitro-1H-imidazol-1-yl)propan-2-yloxycarbonyl]benzoic acid.

The asymmetric unit of the title compound, C(15)H(14)ClN(3)O(6), contains two independent mol-ecules. The imidazole rings are oriented with respect to the benzene rings at dihedral angles of 19.66 (3) and 21.64 (3)°. In the crystal structure, inter-molecular O-H⋯N hydrogen bonds link the mol-ecules into infinite chains.

6,10,16,19-Tetra-oxatrispiro-[4.2.2.4.2.2]nona-deca-ne.

The asymmetric unit of the title compound, C(15)H(24)O(4), contains one half-mol-ecule; a twofold rotation axis passes through the central C atom. The non-planar six- and five-membered rings adopt chair and envelope conformations, respectively. In the crystal structure, inter-molecular C-H⋯O hydrogen bonds link the mol-ecules.

N,N-Dimethyl-3-(1-naphth-yloxy)-3-(2-thien-yl)propan-1-amine.

The title compound, C(19)H(21)NOS, is an inter-mediate for the synthesis of duloxetine hydro-chloride. In the mol-ecular structure, the thio-phene and naphthalene ring systems make a dihedral angle of 87.5°. All bond lengths and angles involving heteroatoms are as expected. In the crystal structure, no classical hydrogen bonds are found.

N-{(E)-4-[(E)-(Dodecyl-imino)meth-yl]benzyl-idene}dodecan-1-imine.

The title compound, C(32)H(56)N(2), was synthesized by the reaction of terephthalaldehyde and dodecan-1-amine. The imines adopt trans conformations, with the two halves of the mol-ecule related to each other by a centre of symmetry.

1-Benzyl-1,4-diazepan-5-one.

The title compound, C(12)H(16)N(2)O, is a diazepane inter-mediate that can be used as an inhibitor of human nitric oxide synthesis. In the mol-ecule, the seven-membered ring has a chair-like conformation and the two rings are approximately perpendicular to one another, with a C-N-C-C torsion angle of 77.8 (4)°. Inter-molecular N-H⋯O hydrogen bonds link the mol-ecules into dimers around a centre of symmetry, with C-H⋯O inter-actions linking the dimers into infinite sheets.

1,4-Bis(benz-yloxy)-2-tert-butyl-benzene.

The title compound, C(24)H(26)O(2), was obtained unintentionally as the product of an attempted synthesis of a new chiral cobalt salen catalyst. There are no classical hydrogen bonds; inter-molecular C-H⋯π stacking inter-actions between aromatic rings help to establish the mol-ecular conformation.