Ergosterol is an economically important metabolite produced by yeast. To improve the production of ergosterol by Saccharomyces cerevisiae YEH56 (pHXA42) from molasses, a cheap and regenerative material, different strategies were applied. First, Plackett-Burman design and central composite design were applied to screen the significant factors in fermentation medium using ergosterol yield (g/L) as the response value. Ergosterol yield reached 371.56 mg/L by using the optimal fermentation medium in shake-flask culture (total sugar in molasses 40 g/L, KH2PO4 1 g/L, K2HPO4 1.86 g/L, CuSO4 x 5H2O 17.5 mg/L, FeSO4 x 7H2O 13.9 mg/L, MgSO4 x 5H2O 12.3 mg/L, corn steep liquor 10 mL/L), which was increased by 29.5% compared with the initial culture. Second, ergosterol yield was increased by 62.1% using a pH-control strategy in a 5-L bioreactor. Third, ergosterol production was improved further by using molasses feeding strategy. After 38 h fermentation, ergosterol yield reached 1 953.85 mg/L, which was 3.2 times of that in batch fermentation. Meanwhile, ergosterol production rate was increased by 42.7% compared with that in the batch culture.
Culture Media ; Ergosterol ; biosynthesis ; Fermentation ; Industrial Microbiology ; Molasses ; Saccharomyces cerevisiae ; genetics ; growth & development ; metabolism

Objective: To study the effect of serum from maintenance hemodialysis(MHD) patients on the expression of intercellular adhesion molecule-1(ICAM-1) in human umbilical vein endothelial cells(HUVEC) cultured in vitro, and the mechanism of endothelial cells dysfunction caused by serum from MHD patients thereof. Methods: HUVEC were incubated for 1 h, 3 h, 6 h and 12 h in RPMI 1640 culture media containing 10 % fetal cattle serum(FCS )(group A), 10% normal human serum (group B) and 10% MHD patient serum (group C) respectively. The immunocytochemical method was used to determine the expression of ICAM-1. Results: There were significant differences in the expression of ICAM-1 in HUVEC among stimulation groups with different serums(P < 0.05). Compared with groups A and B, the expression of ICAM-1 was significantly increased in HUVEC of group C. There was significant difference in the expression of ICAM-1 at the different stimulation times(P < 0.05). There was significant interaction between the expression of ICAM-1 in HUVEC and stimulation groups with different serum and the stimulation times(P < 0.05). Conclusion: The serum from MHD patients with chronic renal failure can promote the expression of ICAM-1 in HUVEC cultured in vitro, thus causing the dysfunction of endothelial cell.

Objective: To observe the effect of acupuncture at points selected from different regions on the positive expression of interstitial cells of Cajal (ICC) and the stem cell factor (SCF) in gastric antrum tissues in diabetic gastroparesis (DGP) rats, and to explore the influence of region-based point selection on the acupoint combination efficacy. Methods: Sixty Sprague-Dawley (SD) rats were randomly divided into a normal group (group A), a model group (group B), a group of Zusanli (ST 36) plus Zhongwan (CV 12) (group C), a group of Zusanli (ST 36) plus Neiguan (PC 6) (group D), and a group of Zusanli (ST 36) plus non-meridian non-acupoint points (group E), based on the random number table (n=12). DGP rat model was established by single intraperitoneal injection of 2% streptozotocin and common diet. After successful modeling, the rats were treated once a day for 4 weeks. Positive ICC and SCF expressions were measured by immunohistochemistry. Results: Compared with group A, the gastrointestinal propulsion rate of group B showed a statistically significant decrease (P<0.05). Compared with group B, the gastrointestinal propulsion rate and the expression of ICC in the gastric antrum were significantly higher in group C, group D and group E, and the between-group differences were statistically significant (P<0.05); the expression of C-kit protein in group C was statistically significantly higher than that in group D and group E (P<0.05). The expression of SCF protein was significantly increased in group C than in group B, and the difference was statistically significant (P<0.05). Conclusion: Acupuncture can improve the symptoms of delayed gastric emptying in DGP model rats, and regulate the expression of ICC and SCF in gastric antrum tissues. The effect of Zusanli (ST 36) plus Zhongwan (CV 12) in the gastric region is superior to that of the Zusanli (ST 36) plus distal Neiguan (PC 6) or non-meridian non-acupoint point, indicating that region-based point selection is the key factor affecting the effect of acupoint combination.

Yeast flocculation is described as a reversible, asexual and calcium dependent process, in which cells adhere to form flocs by interaction of specific cell surface proteins named flocculins on yeast cells with mannose residues present on the cell wall of adjacent yeast cells. Yeast flocculation provides a very economical and convenient pathway for separation of yeast cells from the fermentation broth or removal of heavy metal ions from effluent. A large number of tandem repeats have been found in genes encoding flocculins, which not only have great regulatory effect on the structure and function of flocculins, generating the diversity of flocculation characteristics, but lead to genetic instability in flocculation as well for driving slippage and recombination reactions within and between FLO genes. Here, the research progress in effect of variation of tandem repeats in FLO genes on flocculation characteristics and genetic stability were reviewed to direct and promote the controllable application of flocculation in industrial fermentation process and environmental remediation.
Fermentation ; Flocculation ; Mannose ; Membrane Proteins ; genetics ; Saccharomyces cerevisiae ; genetics ; growth & development ; Saccharomyces cerevisiae Proteins ; genetics ; Tandem Repeat Sequences

The heterologously expressed L1 protein of human papilomavirus 16 can assembly into virus-like particles (VLPs), which has been used as prophylactic vaccine for cervical carcinoma. To express L1 protein in Hansenula polymorpha, we analyzed the codon usage of the native gene of L1 protein and redesigned the encoding sequence according to the codon bias of H. polymorpha. We used assembly PCR to synthesize the native gene HPV16L1-N and the codon optimized gene HPV16L1. The synthesized genes were cloned into pMOXZa-A vector to generate plasmids pMOXZ-HPV16N and pMOXZ-HPV16. The expression cassettes MOXp-HPV16L1(N)-AOXTT were cloned into YEp352 vector and transferred into H. polymorpha. After methanol inducement, the expression of L1 protein in H. polymorpha was detected from the codon optimized gene HPV16L1 rather than the native gene HPVI6L1-N. The parameters for induced cultivation for strain HP-U-16L with HPV16L1 were investigated in shaking flask cultures. After induced cultivation in YPM (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 12 h at 37 degrees C for 72 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by H. polymorpha.
Capsid Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Codon ; genetics ; Genetic Vectors ; genetics ; Human papillomavirus 16 ; genetics ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

Objective:To evaluate the safety and efficacy of ciprofol and propofol for gynecological surgeries with general anesthesia through a meta-analysis.Methods:Electronic databases including PubMed, Embase, Cochrane, Web of Science, China National Knowledge Infrastructure, Wanfang Data, China Biomedical Literature Database, and China Science and Technology Journal Database were searched for randomized controlled trials comparing the safety and efficacy of ciprofol and propofol in gynecological surgeries with general anesthesia from inception to May 2023. Meta-analysis was performed using Revman 5.4 software.Results:Six randomized controlled trials were included, involving 741 patients, of which 371 received ciprofol and 370 received propofol. Compared with propofol group, the emergence time was significantly prolonged, the difference in mean arterial blood pressure, systolic blood pressure and diastolic blood pressure before and after anesthesia induction was decreased, and the incidence of injection pain, respiratory depression, body movement and hypotension was decreased in ciprofol group ( P<0.05). There were no significant differences between the two groups in terms of time of successful anesthesia induction, difference in BIS values and heart rate before and after anesthesia induction, and incidence of tachycardia, bradycardia and hypertension ( P>0.05). Conclusions:Ciprofol is comparable to propofol in terms of efficacy and has better safety than propofol when used in gynecologic surgeries with general anesthesia.

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.