Objective:To investigate the effect of enteral nutrition support on quality of life and therapeutic effect in patients with metastatic colorectal cancer.Methods:A total of 100 patients with metastatic colorectal cancer admitted to Anqing Petrochemical Hospital from Jan. 2020 to Dec. 2023 were selected and divided into parenteral nutrition group and enteral+parenteral nutrition group with 50 cases each using random number table method. The parenteral nutrition group received parenteral nutrition, and the parenteral + parenteral nutrition group was supplemented with enteral nutrition. The therapeutic effect, nutritional indexes, intestinal flora, survival time and quality of life before and after treatment were compared between the two groups.Results:The effective rate of enteral + off-site nutrition group was 76.00%, that of parenteral nutrition group was 54.00%, and that of enteral + off-site nutrition group was higher than that of parenteral nutrition group ( P<0.05). There were no significant differences in nutritional indexes, flora imbalance grade or QLQ-C30 score between the two groups before intervention ( P>0.05). After the intervention, the albumin and total protein in the enteral + parenteral nutrition group were (38.76±6.02) g/L, (64.09±6.71) g/L, the number of normal microbiota disorder cases was 46, the survival time was (17.055±4.33) months, the physical function score was (74.59±7.55) points, and the emotional function score was (78.94±7.96) points, cognitive function score (88.95±9.03) points, role function score (85.49±8.61) points, social function score (81.45±8.27) points. In the parenteral nutrition group, the albumin was (34.51±5.47) g/L, the total protein was (58.91±6.55) g/L, the number of normal microbiota disorder cases was 33, the survival time was (12.48±3.59) months, the physical function score was (67.21±6.81) points, and the emotional function score was (73.55±7.78) points, cognitive function score was (83.47±8.55) points, role function score was (80.14±8.26) points, social function score was (76.93±7.827) points, and enteral + off-site nutrition group was higher than parenteral nutrition group ( P<0.05). In the enteral + off-site nutrition group, the transferrin was (1.45±0.57) g/L, and there were 2 cases of class II flora dysregulation and 2 cases of class III flora dysregulation; in the off-site nutrition group, the transferrin was (1.71±0.61) g/L, and the number of class II flora dysregulation was 8 cases and the number of class III flora dysregulation was 9 cases. Enteral + off-site nutrition group was lower than parenteral nutrition group ( P<0.05) . Conclusion:Enteral + parenteral nutrition support Enteral nutrition support can help improve the treatment effect of metastatic colorectal cancer, improve the nutritional status and survival time of patients, and improve the quality of life of patients.

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

Chemically defined medium is widely used for culturing mouse embryonic stem cells (mESCs), in which N2B27 works as a substitution for serum, and GSK3β and MEK inhibitors (2i) help to promote ground-state pluripotency. However, recent studies suggested that MEKi might cause irreversible defects that compromise the developmental potential of mESCs. Here, we demonstrated the deficient bone morphogenetic protein (BMP) signal in the chemically defined condition is one of the main causes for the impaired pluripotency. Mechanistically, activating the BMP signal pathway by BMP4 could safeguard the chromosomal integrity and proliferation capacity of mESCs through regulating downstream targets Ube2s and Chmp4b. More importantly, BMP4 promotes a distinct in vivo developmental potential and a long-term pluripotency preservation. Besides, the pluripotent improvements driven by BMP4 are superior to those by attenuating MEK suppression. Taken together, our study shows appropriate activation of BMP signal is essential for regulating functional pluripotency and reveals that BMP4 should be applied in the serum-free culture system.
Animals ; Bone Morphogenetic Protein 4/metabolism* ; Cell Differentiation ; Chromosomal Instability ; Endosomal Sorting Complexes Required for Transport ; Mice ; Mitogen-Activated Protein Kinase Kinases/metabolism* ; Mouse Embryonic Stem Cells/cytology* ; Pluripotent Stem Cells/cytology* ; Signal Transduction ; Ubiquitin-Conjugating Enzymes

OBJECTIVE:To study the influence of oscillator on the cleanliness of class 100 clean bench in PIVAS. METH-ODS:Using sedimentated bacteria and the number of dust particle as index,in common drug configuration room,antibiotics con-figuration room and risk drugs configuration room including biological safety cabinet and horizontal laminar flow,the cleanliness of class 100 clean bench were monitored when oscillator was set at clean bench and different positions in work and non-working state. RESULTS & CONCLUSIONS:In working and non-working state of oscillator,there was no difference in sedimentated bacteria and the number of dust particle which was in line with the requirements of 2010 edition of GMP,i.e. the application and location of oscillator didn't influence the cleanliness of class 100 clean bench. From a view of safety,it is suggested to place the oscillator in the left(or right)posterior wall of clean table when biological safety cabinets is used to dispense antibiotic and risk drugs.

Nerve agent not only inhibit acetylcholinesterase ( AChE) at an early stage, but also induce prolonged and progressive neuroinflammation and delayed neurodegeneration.Recently, the US National Institute of Health ( NIH) has sponsored some major programs of toxic mechanisms and treatment of nerve agents, which aims at the development of quick and effective treatment to acute intoxication and delayed effect.The experimentally effective new antidotes mainly include AChE-targeting drugs, broad-spectrum reactivators and scavengers, antiinflamatory and nerve protection drugs.

Objective To compare the changes in energy metabolism in 2-chloroethyl ethryl sulfide(CEES)-poisoned bronchial epithelial cell 16HBE cultured in media at different glucose concentrations .Methods Bronchial epithelial cell 16HBE was cultured in high (4.5 mg/ml) or low (1.1 mg/ml) glucose medium and exposed to a sulfur mustard simulant CEES of 0.2, 0.5, 1.0 mmol/L.Cell growth and cytotoxicity were tested using MTS .ATP, ADP and AMP were detected by HPLC and the value of ATP/ADP, total adenine nucleotides ( TAN) and energy charge ( EC) was subsequently calculat-ed.Mitochondrial oxidative phosphorylation-related proteins, COX-10 and ISCU, were detected using Western blotting . Rhodamine 123 was applied to detect the mitochondrial membrane potential using flow cytometry .Results Low glucose accelerated the growth and energy metabolism of 16HBE cells in regular culture , and the contens of ADP , TAN, COX-10 and ISCU in low glucose group were significantly higher than those in high glucose group .CEES exposure (≥0.5 mmol/L) significantly affected cell viability in both high and low glucose groups , with significant difference between the two groups exposed to 1.0 mmol/L CEES.In high glucose group, 24 h after 0.5 or 1.0 mmol/L CEES exposure, the contents of ATP, ADP and TAN were significantly increased , while ATP/ADP and EC decreased .In low glucose group , ADP, AMP and TAN significantly decreased, while ATP/ADP and EC increased 24 h after 1.0 mmol/L CEES exposure.The mi-tochondrial membrane potential (MMP) also changed differently after 0.5 mmol/L CEES exposure.MMP in high glucose group marginally increased at 3 h, and significantly increased at 8-12 h (P<0.05), and returned to normal at 24 h. MMP in low glucose group showed a transient decrease at 5 h (P<0.01), and back to normal at 8 h.The protein levels of COX-10 and ISCU were significantly increased in high glucose group 24 h after 0.5-1.0 mmol/L CEES exposure , but sig-nificantly decreased in low one 24 h after 1.0 mmol/L CEES exposure .Conclusion When 16HBE is cultured at a high or low glucose concentration , the cell growth, stress responses and energy metabolism including MMP , COX-10, ISCU and ATP production are in different status before or after CEES exposure .High glucose could protect against CEES exposure .

Objective To explore the effect of 2-chloroethyl ethyl sulfide(CEES) poisoning on keratinocyte migration and the regulatory role of microRNA(miR)-34a.Methods MTS was used to detect the viability of cells exposed to CEES in order to select an appropriate dose of CEES exposure in this in vitro model.The protein level of keratin 5 and keratin 10 was detected to assess cell differentiation status .Scratch assay was applied to evaluate cell migration ,and miR-34a silencing in keratinocytes was achieved by transfecting chemically synthesized miR-34a specific miRNA inhibitor.t-ERK1/2 and p-ERK1/2 levels closely related to cell migration were detected using Western blotting .Results An in vitro CEES exposure model of keratinocytes was established at the optimal concentration of 0.5 mmol/L CEES in the viability test , and this dose was chosen to evaluate cell migration changes .The migration of cells was significantly inhibited 24 h after CEES exposure , accompanied by no changes in morphology and keratin 5/10 levels.Silencing of miR-34a significantly increased the migration of cells exposed to CEES , which could be blocked by adding 5 μmol/L U0126 , an ERK1/2 phosphorylation selective inhibitor.Conclusion Silencing of miR-34a can significantly increase keratinocyte migration and partially reverse the inhibition of CEES-caused migration , which could be mediated by ERK 1/2 pathway activation .

OBJECTIVETo investigate the habitat processing method of Forsythiae Fructus based on the different indexes, and to choose the best habitat producing process.

METHODThe habitat producing process was researched by the L9(3(4)) orthogonal and the single factor test. The contents of phillyrin and forsythiaside A were determined by HPLC.

RESULTThe contents of phillyrin and forsythiaside A from Forsythiae Fructus processed by evaporating were 1.33-3.05, and 9.71-44.82 mg x g(-1), respectively. The contents of phillyrin and forsythiaside A from Forsythiae Fructus processed by cooking were 4.63-5.46 mg x g(-1), and 40.20-64.84 mg x g(-1), respectively.

CONCLUSIONWith phillyrin and forsythiaside A as evaluation indexes, cooking method is superior to evaporate method while it is superior to dry method. It is conductive to the preservation and stability of phillyrin and forsythiaside A that add 4 times water of material volum and boil 10 min.

Chromatography, High Pressure Liquid ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Ecosystem ; Forsythia ; chemistry ; Glucosides ; analysis ; chemistry ; Glycosides ; analysis ; chemistry

The integrity and transparency of cornea plays a key role in vision. Limbal Stem Cells (LSCs) are precursors of cornea, which are responsible for self-renewal and replenishing corneal epithelium. Though it is successful to cell replacement therapy for impairing ocular surface by Limbal Stem Cell Transplantation (LSCT), the mechanism of renew is unclear after LSCT. To real time follow-up the migration and differentiation of corneal transplanted epithelial cells after transplanting, we transfected venus (a fluorescent protein gene) into goat LSCs, selected with G418 and established a stable transfected cell line, named GLSC-V. These cells showed green fluorescence, and which could maintain for at least 3 months. GLSC-V also were positive for anti-P63 and anti-Integrinbeta1 antibody by immunofluorescent staining. We founded neither GLSC-V nor GLSCs expressed keratin3 (k3) and keratinl2 (k12). However, GLSC-V had higher levels in expression of p63, pcna and venus compared with GLSCs. Further, we cultivated the cells on denude amniotic membrane to construct tissue engineered fluorescent corneal epithelial sheets. Histology and HE staining showed that the constructed fluorescent corneal epithelial sheets consisted of 5-6 layers of epithelium. Only the lowest basal cells of fluorescent corneal epithelial sheets expressed P63 analyzed by immunofluorescence, but not superficial epithelial cells. These results showed that our constructed fluorescent corneal epithelial sheets were similar to the normal corneal epithelium in structure and morphology. This demonstrated that they could be transplanted for patents with corneal impair, also may provide a foundation for the study on the mechanisms of corneal epithelial cell regeneration after LSCT.
Animals ; Cell Culture Techniques ; methods ; Cell Line ; cytology ; Epithelium, Corneal ; cytology ; metabolism ; Fluorescent Antibody Technique, Indirect ; Goats ; Limbus Corneae ; cytology ; Stem Cell Transplantation ; Stem Cells ; cytology

Objective To explore the changes of interleukin-4,8 and TPK in PC_(12) cells exposed to hypoxia and soman intoxication.Methods PC_(12) cells were randomized into 4 groups: control,hypoxia only,soman only,hypoxia plus soman.The activity of IL-4,IL-8 was measured by ELISA and the activity of tyrosine protein kinase in cells were detected by RIA.Results The activity of IL-4,IL-8 in PC_(12) cell supernatant and TPK in PC_(12) cell cytoplasm,nucleus were increased in 2 h and reached the highest at 12 h,decreased at 24 h in soman group,correspondingly the activity of IL-4,IL-8 in PC_(12) cell supernatant and TPK in PC_(12) cell cytoplasm,nucleus were increased at 2 h and reached the highest at 6 h,decreased at 12 h in soman plus hypoxia group,all higher than that of control.Conclusion TPK,cytokine and its correlated signal pathway lead to an important modulatory role in the brain damage after combined soman and hypoxia injury.