Biologically produced and metal-organic framework delivered dual-cut CRISPR/Cas9 system for efficient gene editing and sensitized cancer therapy.

Manipulation of the lactate metabolism is an efficient way for cancer treatment given its involvement in cancer development, metastasis, and immune escape. However, most of the inhibitors of lactate transport carriers suffer from poor specificity. Herein, we use the CRISPR/Cas9 system to precisely downregulate the monocarboxylate carrier 1 (MCT1) expression. To avoid the self-repairing during the gene editing process, a dual-Cas9 ribonucleoproteins (duRNPs) system is generated using the biological fermentation method and delivered into cells by the zeolitic imidazolate framework-8 (ZIF-8) nanoparticles, enabling precise removal of a specific DNA fragment from the genome. For efficient cancer therapy, a specific glucose transporter 1 inhibitor (BAY-876) is co-delivered with the duRNPs, forming BAY/duRNPs@ZIF-8 nanoparticle. ZIF-8 nanoparticles can deliver the duRNPs into cells within 1 h, which efficiently downregulates the MCT1 expression, and prohibits lactate influx. Through simultaneous inhibition of the lactate and glucose influx, BAY/duRNPs@ZIF-8 prohibits ATP generation, arrests cell cycle, inhibits cell proliferation, and finally induces cellular apoptosis both in vitro and in vivo. Consequently, we demonstrate that the biologically produced duRNPs delivered into cells by the nonviral ZIF-8 carrier have expanded the CRISPR/Cas gene editing toolbox and elevated the gene editing efficiency, which will promote biological studies and clinical applications. STATEMENT OF SIGNIFICANCE: The CRISPR/Cas9 system, widely used as an efficient gene editing tool, faces a challenge due to cells' ability to self-repair. To address this issue, a strategy involving dual-cutting of the genome DNA has been designed and implemented. This strategy utilizes biologically produced dual-ribonucleoproteins delivered by a metal-organic framework. The effectiveness of this dual-cut CRISPR-Cas9 system has been demonstrated through a therapeutic approach targeting the simultaneous inhibition of lactate and glucose influx in cancer cells. The utilization of the dual-cut gene editing strategy has provided valuable insights into gene editing and expanded the toolbox of the CRISPR/Cas-based gene editing system. It has the potential to enable more efficient and precise manipulation of specific protein expression in the future.

[Traditional application and modern research progress on new foreign medicinal resources].

Chinese medicinal resources are the material basis for the survival and development of traditional Chinese medicine(TCM)and the sustainable development of Chinese medicinal resources is also an important project for the modernization of TCM in China. With the increasing demand for Chinese medicinal resources in China, over-exploitation has destroyed Chinese medicinal resources, resulting in a shortage of many natural medicinal resources in China and making the sustainable development of TCM in trouble. The introduced new foreign medicinal resources have become effective supplement and replacement for Chinese medicinal resources to some extent. However, the development and utilization of new foreign medicinal resources in China are different. To fully understand the development of new foreign medicinal resources in China, this paper, taking 43 new foreign medicinal resources such as Acacia nilotica as objects, sorted out the introduction forms and policies of new foreign medicinal resources, overviewed its current development status in China, summarized the application experience of new foreign medicinal resources in the place of origin, as well as the research progress and problems of new foreign medicinal resources in China and abroad, and analyzed the research situation, which can enrich Chinese medicinal resources and other uses, promote the sustainable development of Chinese medicinal resources, and provide ideas for further development and research of new foreign medicinal resources.

[Advances in mechanism of traditional Chinese medicine in inhibiting angiogenesis in ovarian cancer].

Ovarian cancer is one of the three major cancers in gynecology. Ovarian cancer has insidious symptoms in its early stages and mostly has progressed to advanced stages when detected. Surgical treatment combined with chemotherapy is currently the main treatment, but the 5-year survival rate is still less than 45%. Angiogenesis is a key step in the growth and metastasis of ovarian cancer. The inhibition of ovarian cancer angiogenesis has become a new hotspot in anti-tumor targeted therapy, which has many advantages such as less drug resistance, high specificity, few side effects, and broad anti-tumor spectrum. Modern research has confirmed that traditional Chinese medicine(TCM) can inhibit tumor angiogenesis by inhibiting the expression of pro-angiogenic factors, up-regulating the expression of anti-angiogenic factors, inhibiting the proliferation of vascular endothelial cells, reducing the density of tumor microvessels, and regulating related signaling pathways, with unique advantages in the treatment of ovarian cancer. This paper presented a review of the role of TCM in inhibiting ovarian cancer angiogenesis in order to provide references for the optimization of clinical ovarian cancer treatment strategies.

Changes in soil total, microbial and enzymatic C-N-P contents and stoichiometry with depth and latitude in forest ecosystems.

Soil microorganisms and their extracellular enzymes are key factors determining the biogeochemical cycles of carbon (C), nitrogen (N) and phosphorus (P). Relevant studies mainly focus on surface soils (0-20 cm), while deep soils (>20 cm) are often neglected, let alone comparing multiple ecosystems simultaneously. In this study, we studied the latitudinal (19-48°N) and vertical (0-100 cm) patterns of soil total, microbial and enzymatic C-N-P contents and ratios (stoichiometry) in eight temperate, subtropical and tropical forest ecosystems in eastern China. We found that the C-N-P contents and their stoichiometry in soil, microbial biomass and extracellular enzymes all varied significantly with depth and latitude. Soil total C, N and P declined with depth, as did microbial biomass and enzyme activity, while microbial and enzymatic C:N ratios showed increasing or no trend with increasing soil depth. Moreover, soil total and microbial C-N-P contents in surface soils (0-20 cm) showed positive correlations with increasing latitude, and such correlations tended to be weaker or disappeared in deep soils (>20 cm). Overall, changes in total, microbial and enzymatic C-N-P contents and ratios among latitudes suggested a shift from relative N limitation in the north to relative P limitation in the south.

Effects of Cd(II) on wastewater biological nitrogen and phosphorus removal.

Short-term and long-term effects of Cd(II) on wastewater biological nitrogen and phosphorus removal were investigated with respect to microorganism abundances, enzyme activities, and polyhydroxyalkanoates (PHAs) and glycogen transformations. Though no obvious effects on wastewater biological nutrient removal were observed after short-term exposure, the long-term exposure of 10 mg L(-)(1) Cd(II) inhibited nitrification and phosphorus uptake. Compared with the absence of Cd(II), the presence of 10 mg L(-1) of Cd(II) decreased total nitrogen and phosphorus removal efficiencies from 97% and 98% to 88% and 18%, respectively. Mechanism studies revealed that Cd(II) affected the transformations of intracellular PHAs and glycogen, and the activities of oxidoreductase and polyphosphate kinase, resulted in the decrease of nitrite oxidizing bacteria and polyphosphate accumulating organisms abundance, which might be the major reason for the negative effects of long-term exposure to 10 mg L(-1) Cd(II) on biological nitrogen and phosphorus removal.

Water-soluble constituents of the heartwood of Streblus asper.

A new lignan, (7'R,8'R)-threo-strebluslignanol-2-O-beta-D-glucopyranoside, along with 8 known compounds (2-9) were isolated from the water-soluble part of the MeOH extract of the heartwood of Streblus asper. Their structures were elucidated through various spectroscopic methods, including 1D NMR (1H NMR, 13C NMR), 2D NMR (HMQC, HMBC, and NOESY), and HRMS. The stereochemistry at the chiral centers was determined using the CD spectrum, as well as analyses of coupling constants and optical rotation data. In the preliminary bioassay, the isolated compounds did not show anti-HBV activities in vitro using the HBV transfected HepG2.2.15 cell line.

Anti-HBV activities of Streblus asper and constituents of its roots.

The extracts from leaves, heartwood, barks and roots of the Streblus asper were investigated for anti-HBV activities, separately. The results suggested that the MeOH extracts of the heartwood, barks, and roots exhibited good anti-HBV activities. Further investigations displayed that ethyl acetate and n-butanol soluble parts of their MeOH extracts showed more significant anti-HBV activities. Moreover, a new lignan, together with 11 known compounds, was isolated from n-butanol-soluble part of MeOH extract of the roots of S. asper. The structures were elucidated by spectroscopic methods, including 1D NMR ((1)H NMR, (13)C NMR), 2D NMR (HMQC, HMBC) and HR-EI-MS experiments. Compounds 1-3 were evaluated for their anti-HBV activities. Honokiol showed significant anti-HBV activity with IC(50) values of 3.14µM and 4.74µM for HBsAg and HBeAg with no cytotoxicity respectively, while lamivudine (3TC, positive control) exhibited weak anti-HBV activity with IC(50) values of 11.81µM and 25.80µM for HBsAg and HBeAg respectively.

Hypoglycemic effects and constituents of the barks of Cyclocarya paliurus and their inhibiting activities to glucosidase and glycogen phosphorylase.

The various fractions of the barks of Cyclocarya paliurus were systematically tested for hypoglycemic effects in alloxan-induced diabetic rats. The results showed that the chloroform fraction of the 75% EtOH extract of the barks of this plant exhibited significant blood sugar reducing activity, most of which were significantly higher than that of positive-drug metformin hydrochloride. A new compound, together with nine known compounds, was isolated from the most active fraction. The structure elucidation was based on spectroscopic methods, including two-dimensional NMR experiments (¹H-¹H COSY, HMQC, and HMBC). All of the isolates were evaluated for their α-glycosidase and glycogen phosphorylase inhibitory activities.

[Study on vascular morphology of integument tissues of the channel area in the leg].

OBJECTIVE: To observe the distribution of the blood vessels in the integument tissue of the channel area of legs. METHODS: The integument tissue of the lower limbs in the 12 cadavers were dissected with macro-and micro-dissection, radiographical technique of systemic artery and technique of image pattern analysis to observe and analyze the origins, branches and anastomoses in the integument tissues along the channels of legs. RESULTS: The distributional density of the blood vessels in the integument tissues of legs along the channel area of the three-yin meridians of the foot, the Gallbladder Meridian, and the Urinary Bladder Meridian was higher than that in the other parts. They formed an obvious nutrient vascular chain on the arteriogram. The distributional density in the channel area of the Stomach Meridian was not obviously increased and the obvious nutrient vascular chain could not be seen. CONCLUSION: An obvious nutrient vascular chain is formed in the integument tissue along the channel area of legs except the Stomach Meridian.