Downregulated FGFR3 Expression Inhibits In Vitro Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells of Mice with TBXT Gene Mutation.

We studied the effect of fibroblast growth factor receptor 3 (FGFR3) inhibitor BGJ-398 on the differentiation of bone marrow mesenchymal stem cells (BM MSC) into osteoblasts in wild type (wt) mice and in animals with mutation in TBXT gene (mt) and possible differences in the pluripotency of these cells. Cytology tests showed that the cultured BM MSC could differentiate into osteoblasts and adipocytes. The effect of different BGJ-398 concentrations on the expression of FGFR3, RUNX2, SMAD1, SMAD4, SMAD5, SMAD6, SMAD7, and SMAD8 were studied by quantitative reverse transcription PCR. The expression of RUNX2 protein was evaluated by Western blotting. BM MSC of mt and wt mice did not differ in pluripotency and expressed the same membrane marker antigens. BGJ-398 inhibitor reduced the expression of FGFR3 and RUNX2. In BM MSC from mt and wt mice have similar gene expression (and its changing) in FGFR3, RUNX2, SMAD1, SMAD4, SMAD5, SMAD6, SMAD7, and SMAD8 genes. Thus, our experiments confirmed the effect of decreased expression of FGFR3 on osteogenic differentiation of BM MSC from wt and mt mice. However, BM MSC from mt and wt mice did not differ in pluripotency and are an adequate model for laboratory research.

Antibacterial spectrum of four compounds from yeasts in koumiss.

Koumiss has beneficial therapeutic effects on bacterial diseases. Four antibacterial com- pounds from yeasts (Kluyveromyces marxianus and Saccharomyces cerevisiae) in koumiss were evaluated for their antibacterial effects against three Gram-negative bacteria, three Gram-positive bacteria and five pathogenic Escherichia coli strains. The antibacterial compounds from yeasts in koumiss were extracted, and their main components were determined. The inhibition zones were analyzed, and their minimum inhibition concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined. Aqueous phases of Kluyveromyces marxianus and Saccharomyces cerevisiae at pH 2.0 and 8.0 produced larger inhibition zones than those in other phases, and then antibacterial compounds from K. marxianus (K2, pH=2.0; K8, pH=8.0) and S. cerevisiae (S2, pH=2.0; S8, pH=8.0) were obtained. Their main components were organic acids and killer toxins. K2 had more propanoic acid and S2 had more oxalic acid than others. The inhibition zones of K2, K8, S2 and S8 against three Gram-negative bacteria and three Gram-positive bacteria were 12.03-23.30 mm, their MICs were 0.01-0.13 g/mL, and MBCs were 0.03-0.50 g/mL. Meantime, the inhibition zones of K2, K8, S2 and S8 against five pathogenic E. coli were 16.10-25.26 mm, their MICs were 0.03-0.13 g/mL, and MBCs were 0.13-1.00 g/mL. These four antibacterial compounds from yeasts in koumiss had broad antibacterial spectrum. In addition, K2 and S2 were better than K8 and S8.

The detection of SAS1B in serum provides clues for early diagnosis of thyroid cancer.

OBJECTIVE: The incidence of thyroid cancer is rising globally. Most patients progress slowly, but some patients develop lymph node and distant metastasis earlier, and their prognosis is poor. Therefore, early diagnosis and warning of malignancy are very meaningful for such patients. SAS1B gene is a newly discovered protein expressed on the surface of mature egg cells and has metalloendopeptidase activity. We aimed at exploring whether SAS1B is involved in the occurrence of thyroid cancer, and at providing evidence for early diagnosis and targeted therapy of thyroid cancer. PATIENTS AND METHODS: In this study, a rabbit anti-human SAS1B polyclonal antibody was prepared by gene recombination technology. The indirect ELISA method was used to detect the SAS1B protein expression in the serum of 69 patients with thyroid cancer and 55 normal controls, and the relevant pathological factors were analyzed. Immunohistochemistry and PCR technology were used to investigate the expression levels of SAS1B protein and mRNA in 30 thyroid cancer tissues and 23 control thyroid tissues. RESULTS: The titer of SAS1B recombinant antibody was 1:51200. The expression of SAS1B in the serum of patients with thyroid cancer was higher than that in the normal control group (p<0.01). The antibody had a good sensitivity in serum detection of cancer patients (p=0.008<0.01), the linear regression analysis result was that the expression of SAS1B gene was related to tumor envelope invasion and lymph node metastasis (p=0.003<0.01, p=0.003<0.01), and it was irrelevant to the patient's gender, age, tumor mass size, number of cancer foci, pathological stage, etc. (p>0.05). The results of immunohistochemistry showed that SAS1B protein was mainly located in the cytoplasm and membrane of thyroid cancer cells. The expression intensity in thyroid cancer tissues was higher than that in control tissues (p<0.05), but it was not expressed in normal thyroid tissues. Antibodies showed a good sensitivity that was used to detect thyroid cancer tissues (p=0.000<0.01). The results of ordinary PCR detection using thyroid cancer tissue and control thyroid tissue showed that the amplification products of the three domains (N-terminal, C-terminal and catalytic domain) of the SAS1B gene showed high expression in thyroid cancer tissue. q-PCR results showed that the expression of SAS1B gene in thyroid cancer and control thyroid tissue was higher than that in control group (p<0.05), and the genes of Aurora A and BARD1 related to centrosome replication and DNA replication forks protection during the proliferation were highly expressed in thyroid cancer tissue. The study results suggested that SAS1B was involved in the carcinogenesis of thyroid cancer. The Hum_mPLoc.2.0 software, PSORT Ⅱ software and UniProt software were used to predict that SAS1B protein had secretory protein properties. CONCLUSIONS: The above data indicate that the SAS1B gene is closely related to the process of thyroid cancer and can serve as a good tumor marker that can be used for early diagnosis and early warning of thyroid malignancy.

[Progress in diagnosis and treatment of ipsilateral femoral neck and shaft fracture].

Ipsilateral femoral neck and shaft fractures are rare injuries, which are often caused by high-energy trauma and combined with multiple injuries, such as thoracic and abdominal injury, head injuries, and fractures of other sites.Delayed or missed diagnosis of the ipsilateral femoral neck fracture often occurs.When patients with femoral shaft fractures caused by high-energy trauma are admitted into hospital, physical examination should be conducted carefully.In addition to femoral shaft fractures, radiographs of the ipsilater hip and knee joints should been taken, simultaneously taking into consideration the potential effect of anteversion angle on the demonstration of femoral neck fracture.Computed tomograph and magnetic resonance imaging are advised to perform if necessary to facilitate early and accurate diagnosis of ipsilateral femoral neck fracture.Comprehensive evaluation should be done based on age, physical condition, associated injuries as well as fracture site, classification and injury severity.Accordingly, proper and reasonable surgical plan is made.During the operation, anatomical reduction of the fractures, especially femoral neck fractures, should be achieved, and then fixed with appropriate internal implants.Besides, attention should also be paid to the treatment of associated injuries as well as the prevention and management of complications.

Expression of the RORα gene in Inner Mongolian cashmere goat hair follicles.

The expression of retinoid-acid-related orphan receptor α (RORα) was evaluated at the mRNA level using real-time polymerase chain reaction (qRT-PCR), and its expression localization was determined by in situ hybridization of adult Inner Mongolian cashmere goats at different times of the year. In situ hybridization demonstrated that RORαwas expressed in secondary hair follicles of the hair shaft, inner root sheath, outer root sheath, medulla, and other parts that are target organs of the RORαreceptor gene. qRT-PCR results showed that there was no significant difference in the RORa mRNA abundance in February, April, August, and October (P > 0.05), and the only difference occurred in December relative to February, August, and October (P < 0.05). This difference revealed that melatonin possibly promotes cashmere growth through the nuclear receptor RORα. This study provides a good foundation for future studies on the relationship between the melatonin receptor and cashmere growth; in addition, it provides new insights for increased cashmere production and quality.

FasL-induced apoptosis in bovine oocytes via the Bax signal.

The factor associated suicide (Fas) and its ligand (FasL) signaling is an important regulatory pathway of apoptosis in mammalian follicles. However, whether apoptosis in bovine oocytes is regulated by the Fas-FasL signaling pathway remains unknown. In this study, localization of Fas and FasL in immature oocytes and FasL in cumulus cells were examined using immunofluorescence staining. In addition, exogenous FasL was added to an in vitro culture system to investigate apoptotic changes in bovine oocytes, using annexin-V and terminal uridine nick-end labeling staining, and real-time quantitative polymerase chain reaction. In this study, Fas was expressed in immature oocytes, whereas FasL was expressed in cumulus cells, but not in immature oocytes; annexin-V- and terminal uridine nick-end labeling-positive rates of oocytes treated with 2, 10, or 50 ng/mL FasL were higher than those of control oocytes (P < 0.05); and oocytes from the three treatment groups had higher expression levels of Fas and B cell lymphoma/leukemia-2 associated X than those in the control group (P < 0.05). Taken together, we concluded that the Fas-FasL signaling pathway was involved in regulation of bovine oocyte apoptosis, perhaps related to B cell lymphoma/leukemia-2 associated X upregulation.