OBJECTIVE To study the relationship of the expression of PLOD2 protein in laryngeal carcinoma and the clinicopathological features of patients. METHODS The expression of PLOD2 in paraffin-embedded specimens of 114 patients with laryngeal carcinoma was detected by immunohistochemistry. The relationship between the expression of PLOD2 and clinicopathological features was analyzed by χ2 t est, s urvival a nalysis b y K aplan-Meier method, and multivariate analysis of Cox proportional hazard model. The fresh frozen specimens of 8 patients randomly selected from the patients were detected by realtime quantitative polymerase chain reaction and Western blotting for the expression of PLOD2 in tumor tissues and adjacent normal tissues. RESULTS PLOD2 protein was associated w ith c linical s tage a nd T s tage(P <0.05). The expression level of PLOD2 protein in laryngeal squamous cell carcinoma was higher than that in adjacent normal tissue(P <0.05). Kaplan-Meier survival analysis showed that low expression of PLOD2 was associated with patient survival rate(χ2=12.484, P <0.001). Multivariate Cox regression analysis showed that PLOD2 protein expression and M stage were independent risk factors for laryngeal cancer growth (P value, both <0.05). CONCLUSION The level of POLD2 protein expression was positively correlated with clinical stage and T stage. PLOD2 protein is an independent risk factor for the growth of laryngeal cancer. The higher the expression of PLOD2 protein, the lower the prognosis of patients. PLOD2 protein expression may play an important role in the growth and prognosis of laryngeal cancer, and may be a new molecular marker for judging the growth and prognosis of laryngeal cancer.

OBJECTIVE：To study the ef fects of Gegen qinlian decoction （GGQLD）on blood lipid and blood glucose of hyperlipidemia（HLP）model rats ，and to explore its mechanism from the perspective of intestinal flora. METHODS ：Totally 48 rats were randomly divided into blank control group （n＝8）and modeling group （n＝40）. For consecutive 5 weeks，model group was given high-lipid diet to induce HLP model ；blank control group was given routine diet. After modeling ，30 modeling rats were randomly divided into model group ，simvastatin group （positive control ，10 mg/kg），GGQLD high-dose ，medium-dose and low-dose groups （14.85，4.95，1.65 g/kg，by crude drug ），with 6 rats in each group. Blank control group and model group were given constant volume of normal saline intragastrically ；administration groups were given relevant medicine intragastrically ，once a day，for consecutive 11 weeks. At the same time ，each group was continuously given corresponding diet. After the last medication ， body mass and body length of rats were determined ，and Lee ’s index was calculated. Serum levels of TG ，TC，HDL-C，LDL-C and fasting blood glucose （FBG）were determined in rats. DNA of rat caecum content was extracted for 16S rRNA V 3-V4 region sequencing. The Two-part model was used to analyze the correlation between intestinal flora with lipids and blood glucose. RESULTS：After 11 weeks of administration ，compared with blank control group ，the body mass ，body length ，Lee’s index ， serum levels of TC ，TG，HDL-C and FBG of model group were increased significantly （P＜0.05 or P＜0.01），while the level of HDL-C was decreased significantly （P＜0.05）. Compared with model group ，body mass and Lee ’s index and serum levels of TG ， FBG of rats in GGQLD high-dose group ，and serum levels of TC ，TG in GGQLD medium-dose group ，as well as serum level of TG of rats in GGQLD low-dose group was decreased significantly （P＜0.05 or P＜0.01）. Correlation analysis with intestinal flora showed that TC and TG shared 3 operational taxonomic units （OTU），including OTU 559，OTU701 and OTU 135（OTU135 was also shared with FBG ），which were all positively correlated with the level of TC ，TG and FBG （P＜0.01）. The three OTU were annotated as Tyzzerella of Spirillaceae ，Anaerotruncus of Verrucaceae and Peptoclostridium of Streptococcidae ，respectively. High-dose and low-dose GGQLD had a down-regulating effect on Tyzzerella and Anaerotruncus（P＜0.05 or P＜0.01），while had up-regulating effect on Peptoclostridium（P＜0.01）. CONCLUSIONS ：High-dose GGQLD （14.85 g/kg）can effectively reduce the body mass and blood lipid of HLP model rats ，and can prevent the abnormal increase of blood glucose of model rats. The mechanism may be associated with that the reduction of intestinal flora （Tyzzerella，Anaerotruncus）content.

Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.
Pregnancy ; Female ; Animals ; Mice ; Tetraploidy ; Blastocyst ; Embryo, Mammalian ; Cell Differentiation ; Embryonic Development