Determining the safety of a newly developed experimental product is a crucial condition for its medical use, especially for clinical trials. In this regard, four hydrogel-type formulations were manufactured, all of which were based on carbomer (Blank-CP940) and encapsulated with caffeine (CAF-CP940), phosphorus derivatives (phenyl phosphinic (CAF-S1-CP940) and 2-carboxyethyl phenyl phosphinic acids (CAF-S2-CP940)). The main aim of this research was to provide a comprehensive outline of the biosafety profile of the above-mentioned hydrogels. The complex in vitro screening (cell viability, cytotoxicity, morphological changes in response to exposure, and changes in nuclei morphology) on two types of healthy skin cell lines (HaCaT-human keratinocytes and JB6 Cl 41-5a-murine epidermal cells) exhibited a good biosafety profile when both cell lines were treated for 24 h with 150 μg/mL of each hydrogel. A comprehensive analysis of the hydrogel\'s impact on the genetic profile of HaCaT cells sustains the in vitro experiments. The biosafety profile was completed with the in vivo and in ovo assays. The outcome revealed that the developed hydrogels exerted good biocompatibility after topical application on BALB/c nude mice\'s skin. It also revealed a lack of toxicity after exposure to the hen\'s chicken embryo. Further investigations are needed, regarding the in vitro and in vivo therapeutic efficacy and safety for long-term use and potential clinical translatability.

OBJECTIVE: Fat grafting has been widely used for soft-tissue augmentation. External volume expansion (EVE) is a favorable tool for improvement in the rate of fat graft retention. However, few studies have focused on the most appropriate time for its implementation. In this study, BALB/c nude mice were used to investigate the effective time for the implementation of external volume expansion to improve the rate of fat retention.
METHODS: Sixteen mice were divided into four groups, and EVE was performed at different time points before or both before and after fat grafting. Fat tissue from a human donor was injected into the mice following EVE. Visual assessment, micro-computed tomography analysis, and histopathological evaluation were used to assess fat retention.
RESULTS: After 10 weeks, the group that underwent EVE 5 days before fat grafting demonstrated a significantly higher preserved fat volume, as determined by micro-computed tomography (p<0.05). Moreover, the group that received additional EVE after fat grafting exhibited a higher retention rate compared to the groups receiving EVE only before grafting (p<0.05). Histopathological analysis indicated that swelling, edema, and inflammation were more pronounced in the group with EVE immediately before grafting, while angiogenesis and lipogenesis were more active in the group with additional EVE after grafting.
CONCLUSIONS: EVE is a safe and effective approach for improving the rate of fat graft retentions. Furthermore, the timing of external tissue expansion plays a crucial role in fat retention. Based on our animal study, performing EVE immediately before and after fat grafting may be an effective strategy for enhancing the rate of fat graft retentions.

Ent-kaurane diterpenoids were studied as a biologically active ingredient group of Sigesbeckia pubescens (Makino) Makino. Here, five known ent-kaurane diterpenoids were isolated and identified, named ent-16β,17-dihydroxy-kauran-19-oic acid (1), ent-16β,17-dihydroxy-kauran-19-oate (2), ent-18-acetoxy-17-hydroxykauran-19-oic acid (3), ent-16β,17,18-trihydroxy-kauran-19 -oic acid (4), and ent-17-hydroxy-kauran-16βH-19-oic acid (5). Their inhibitory effects of these compounds on MDA-MB-231 breast cancer migration were firstly tested in a chemotaxis invasion assay. Among them, compound 1 (DKA) showed superior inhibitory activities with IC50 value of 1.96 µM. Then, a wound healing assay and BALB/c nude mice were used for further studying the inhibitory activity of DKA on MDA-MB-231 breast cancer migration in vitro and in vivo, respectively. The wound healing assay showed that DKA (1, 5, and 25 μM) can significantly inhibit cell migration and the mouse model of lung metastasis showed that DKA (2.5, 5, and 10 mg/kg) could strongly suppress the lung metastasis of MDA-MB-231 breast cancer cells.

Colorectal cancer (CRC) is one of the most common cancers worldwide and the consumption of a high-calorie diet is one of its risk factors. Calorie restriction (CR) slows tumor growth in a variety of cancers, including colorectal cancer; however, the mechanism behind this remains unknown. In the present study, CR effectively reduced the tumor volume and weight in a xenograft BALB/c male nude mouse model. In addition, tumor immunohistochemistry revealed that the CR group had significantly higher expression of Bax (P<0.001) and significantly lower levels of Bcl2 (P<0.0001) and Ki67 (P<0.001) compared with control group. Furthermore, data from 16S ribosomal (r)RNA sequencing implied that CR was able to reprogram the microbiota structure, characterized by increased Lactobacillus constituent ratio (P<0.05), with amelioration of microbial dysbiosis caused by CRC. Further receiver operating characteristic curves demonstrated that the bacteria Bacteroides [area under the curve (AUC)=0.800], Lactobacillus (AUC=0.760) and Roseburia (AUC=0.720) served key roles in suppression of CRC in the mouse model. The functional prediction of intestinal flora indicated \'cyanoamino acid metabolism\' (P<0.01), \'replication initiation protein REP (rolling circle plasmid replication)\' (P<0.01), \'tRNA G10 N-methylase Trm11\' (P<0.01) and \'uncharacterized protein with cyclophilin fold, contains DUF369 domain\' (P<0.05) were downregulated in CR group. These findings implied that CR suppressed CRC in mice and altered the gut microbiota.

BACKGROUND: This study is to investigate the preventive and therapeutic effect of intraportal oridonin on colorectal cancer liver metastasis (CRCLM).
METHODS: The inhibitory effect of oridonin on HT29 cells was determined by CCK-8 and MTT assays. The preventive and therapeutic effect of intraportal oridonin on CRCLM were investigated by establishing BALb/c nude mice hemispleen models of colon cancer liver metastasis. The microscopic characteristics of tumor tissues were observed by hematoxylin-eosin staining, immunohistochemistry and TUNEL staining. On the other hand, liver function enzymes, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP), were detected to evaluate the hepatotoxicity of intraportal oridonin. The serum levels of tumor markers, including carcinoembryonic antigen (CEA) and α-fetoprotein (AFP), were used to investigate the intervention effect of intraportal oridonin on CRCLM.
RESULTS: Oridonin exerted an inhibitory effect on the proliferation of HT29 cells in vitro. Intraportal oridonin was found to effectively prevent the occurrence and formation of CRCLM, whilst intraportal oridonin can also exert a therapeutic effect on CRCLM. Additionally, liver enzymes testing indicated that intraportal oridonin possesses non-hepatotoxicity, instead can effectively alleviate liver injury caused by tumor. Furthermore, intraportal oridonin was also revealed to decrease the serum levels of AFP and CEA.
CONCLUSIONS: Intraportal oridonin can effectively inhibit the formation of liver metastatic tumor and exert a certain degree of preventive and therapeutic effect on CRCLM. These findings indicate intraportal oridonin to be a promising anti-metastasis agent for CRCLM.

OBJECTIVE: To investigate the difference of tumor formation in different mouse strains bearing patient-derived xenograft of esophageal squamous cell carcinoma(ESCC) and establish a better animal model for preclinical study of individualized treatment of ESCC.
METHODS: The tumor tissues collected from 22 ESCC patients were used to establish tumor-bearing mouse models in B-NDG? (NSG) mice and BALB/c nude mice. The tumor formation rate and tumor formation time were compared between the two mouse models, and HE staining, immunohistochemistry and genome sequencing were carried out to assess the consistency between transplanted tumor tissues in the models and patient-derived tumor tissues.
RESULTS: The tumor-bearing models were established successfully in both NSG mice (50%, 11/22) and BALB/c nude mice (18.18%, 4/22). The average tumor formation time was significantly shorter in NSG mice than in BALB/c nude mice (75.95 vs 91.67 days, P < 0.001). In both of the mouse models, the transplanted tumors maintained morphological characteristics identical to those of patient-derived ESCC tumors. Genetic analysis showed that the xenografts in NSG mice had a greater genetic similarity to the patients\' tumors than those in BALB/c nude mice (P < 0.0001).
CONCLUSIONS: Mouse models bearing xenografts of patient-derived ESCC can be successfully established in both NSG mice and BALB/c nude mice, but the models in the former mouse strain can be more reliable.

BACKGROUND: The effects of low-intensity laser radiation (LILR) on the skin depend on the wavelength and density of the irradiation flux. Moreover, the vast receptor field of the skin facilitates the systemic influence of irradiation on the body.
OBJECTIVE: The objective of the present study was to evaluate the effects of low-intensity laser irradiation (LIRI) of the infrared range with a wavelength of 1270 nm on the skin of mice Balb/cNude.
METHODS: The study was carried out with the use of the linear immunodeficient mice Balb/c nude obtained from the animal house of the Pushchino branch of the Academicians M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Biorganic Chemistry. The animals were irradiated once with a fiber laser of stimulated Raman scattering with a wavelength (λ) of 1270 nm, a power of 1.96 mW during the 2 min exposure at an intensity of 10 mW/cm2, and an energy flux density of 1.2 J/cm2. The histological studies and morphometry of the skin autoptate were performed. In adition, the expression of microRNA-21, -31, -130a, -191, -200c, -205, -218 was determined in the skin. Reverse transcription and real-time PCR reactions with Taq-Man probes and primers were performed on the nucleic acid amplifier CFX96 (\'BioRad\', USA).MicroRNA-191 was chosen as the reference gene. The bioinformation analysis of signaling pathways involving the studied microRNAs was performed using the DJANA miRPath database v.3.0.
RESULTS: A significant increase in the amount of keratinocytes of the basal layer was documented together with diffuse lymphoid-leukocyte infiltration of the interlobular connective tissue of the subcutaneous fat and endomysium after LILR. The pattern of microRNA expression was tissue-specific. An increase in the expression of micro-RNA-31 and-21 in the skin and a multidirectional change in miRNA-200 and -218 levels were shown. The bioinformation analysis showed that miR21 and miR31 were involved in the regulation of such signaling pathways as PI3K-Akt, Jak-STAT, MAPK, and mTOR of importance for carcinogenesis. Also, they have a signaling significance in the development of melanoma, kidney cancer, prostate cancer and malignant glioma.
CONCLUSIONS: The data obtained in this study suggest activation of the tumor cell-specific and basic processes in the skin of immunodeficient Balb/cNude mice under the influence of low-intensity laser radiation with a wavelength of 1270 nm applied at a dose of 1.2 J/m2.
Обоснование. Эффекты воздействия на кожу низкоинтенсивного лазерного излучения (НИЛИ) определяются длиной волны и плотностью потока облучения. Также обширное рецепторное поле кожи обусловливает системное влияние на организм. Цель исследования - изучить влияние НИЛИ инфракрасного диапазона с длиной волны 1270 нм на кожу мышей линии Balb/cNude. Методы. Объектом исследования послужили линейные иммунодефицитные мыши Balb/cNude (питомник лабораторных животных \'Пущино\' филиала ФГБУН \'Институт биоорганической химии им. акад. М.М. Шемякина и Ю.А. Овчинникова\' РАН). Животных однократно облучали волоконным лазером вынужденного комбинационного рассеивания с длиной волны 1270 нм, мощностью 1,96 мВт, экспозицией 2 мин при интенсивности 10 мВт/см2 и плотности потока энергии 1,2 Дж/см2. Проводили гистологическое исследование аутоптата кожи и морфометрию основных структур. В коже также определяли экспрессию микроРНК-21, -31, -130а, -191, -200с, -205 и -218. Реакцию обратной транскрипции и полимеразной цепной реакции в реальном времени с зондами и праймерами TaqMan выполняли на амплификаторе нуклеиновых кислот СFХ96 (\'BioRad\', США). В качестве гена-рефери выбрана микроРНК-191. Биоинформационный анализ сигнальных путей с участием изученных микроРНК проводили с использованием базы данных DJANA miRPath v.3.0. Результаты. Установлено значимое повышение количества кератиноцитов базального слоя на единицу площади, выявлена диффузная лимфо-лейкоцитарная инфильтрация в соединительной ткани подкожной жировой клетчатки и в эндомизии после воздействия НИЛИ. Паттерн экспрессии микроРНК тканеспецифичен. Показаны повышение экспрессии микроРНК-31 и -21 в коже и разнонаправленные изменения микроРНК-200с и -218. Биоинформационный анализ показал, что микроРНК-21 и -31 участвуют в регуляции таких важных сигнальных путей при канцерогенезе, как PI3K-Akt, Jak-STAT, MAPK, mTOR, а также имеют сигнальное значение при развитии меланомы, почечного рака, рака простаты и глиомы. Заключение. Полученные результаты позволяют предположить активацию специфических для опухолевых клеток, а также базовых процессов в коже иммунодефицитных мышей линии Balb/cNude под влиянием НИЛИ с длиной волны 1270 нм в дозе 1,2 Дж/см2.

OBJECTIVE: With the global demand for dairy protein for consumption growing annually, there has been increasing activity in the research field of dairy protein synthesis and production. From a manipulation perspective, it is more difficult to use live cattle for laboratory studies on the production of milk as well as of dairy protein such as casein, as compared with using laboratory animals like rodents. Therefore, we aimed to develop a mouse model of bovine mammary alveolar ducts for laboratory-scale studies. We studied the formation of the bovine mammary gland ductal structure by transplanting the MAC-T bovine alveolar cell line into mice.
RESULTS: MAC-T cells (1×10⁷) were suspended in Matrigel and injected into the dorsal tissue of 8-week-old male BALB/C nude mice. Histological analysis of tissue dissected from the MAC-T cell-transplanted mice after 6 weeks showed the typical morphology of the tubuloalveolar female gland, as well as glands made up of branching ducts that were surrounded by smooth muscle with small alveoli budding off the ducts. In addition, the epithelial markers CK14 and CK18 were expressed within the duct-like structure. Prolactin was detected in the duct interior in these CK14＋ and CK18＋ cells but not in the non-transplanted MAC-T cells.
CONCLUSIONS: These results showed that duct-like tissue had been successfully formed after 6 weeks of transplantation of the CK14＋ and CK18＋ MAC-T cells into mice dorsal tissue. This mouse model will be a useful tool for further research on the bovine mammary gland.