The advances of industrial biotechnology highly depend on the development of industrial bioprocess researches. In China, we are facing several challenges because of a huge national industrial fermentation capacity. The industrial bioprocess development experienced several main stages. This work mainly reviews the development of the industrial bioprocess in China during the past 30 or 40 years: including the early stage kinetics model study derived from classical chemical engineering, researching method based on control theory, multiple-parameter analysis techniques of on-line measuring instruments and techniques, and multi-scale analysis theory, and also solid state fermentation techniques and fermenters. In addition, the cutting edge of bioprocess engineering was also addressed.
Bioengineering ; history ; Bioreactors ; Biotechnology ; Chemical Engineering ; China ; Fermentation ; History, 20th Century ; History, 21st Century

Filamentous fungi are widely used in industrial fermentation. Particular fungal morphology acts as a critical index for a successful fermentation. To break the bottleneck of morphological analysis, we have developed a reliable method for fungal morphological analysis. By this method, we can prepare hundreds of pellet samples simultaneously and obtain quantitative morphological information at large scale quickly. This method can largely increase the accuracy and reliability of morphological analysis result. Based on that, the studies of Aspergillus niger morphology under different oxygen supply conditions and shear rate conditions were carried out. As a result, the morphological responding patterns of A. niger morphology to these conditions were quantitatively demonstrated, which laid a solid foundation for the further scale-up.
Aspergillus niger ; cytology ; Fermentation ; Industrial Microbiology ; Reproducibility of Results

Objective To explore the bone marrow stromal cells,anti-late antigen-4 (VLA-4) antibody (aVLA-4),cytarabine (Ara-C) on the proliferation and apoptosis of leukemia HL-60 cells.Methods The experiment was divided into five groups:HL-60 cells were cultured alone (control group),HL-60 cells and stromal cells group (stromal cells group),HL-60 cells + stromal cells + aVLA-4 (antibody group),HL-60 cells + stromal cells + Ara-C group (drug group),HL-60 cells + stromal cells + aVLA-4 + Ara-C group (antibody +drug group).Cell proliferation or inhibition rate was detected by CCK-8 method,the HL-60 cells apoptosis was detected by flow cytometry.The expression of anti-apoptotic gene bcl-2 in HL-60 cells was determined by Western blot.Results After 24 h and 48 h,treatment,the number of the stromal cells group HL-60 cells were higher than that of the control group with significant difference cultured [(7.2±0.3)×1O5/ml vs (5.3±0.4)×105/ml,(8.4±0.2)×105/ml vs (6.8±0.3)×105/m1,P ＜ 0.001],while the HL-60 cell proliferation inhibition rate [(24.3±2.1) ％,(37.0±2.6) ％,(65.6±3.8) ％] and apoptosis rate [(5.7±0.6) ％,(8.0±0.5) ％,(10.4±0.9) ％,(16.5±0.7) ％] of antibody group,drug group,antibody + drug group were higher than the control group with a difference of statistically significant (P ＜ 0.05),and the increase of antibody + drug group was most obvious.With the decreasing of the bcl-2 protein expression,which was most the decrease of antibody + drug group was most obvious.Conclusion Bone marrow stromal cells can stimulate the proliferation of leukemia cells,aVLA-4 interference the interaction between stromal cells and leukemia cells can enhance the chemosensitivity of leukemia cells to Ara-C.

Objective To evaluate DTI for early diagnosis and prognosis of diffuse axonal injury (DAI)in the moderate and sever traumatic brain injury (sTBI).Methods 30 patients with sTBI and 30 healthy controls were studied with T1 WI and DTI.The frac-tional anisotropy(FA)and ADC were quantified from different regions of interest(ROI)including the genu of corpus callosum(gCC), the trunk of CC,the splenium of CC(sCC)and the integrity CC.The FA and ADC of the same R0I was compared between TBI group and control group,and FA and ADC of the different parts of CC were compared in TBI group.Correlations between the FA or ADC and coma duration of the TBI patients were analyzed by Pearson linear correlation.Results Compared with control group,the FA value decreased significantly in each ROI,while ADC was reversed.The FA or ADC of the different ROI were correlated with co-ma duration in TBI group.Conclusion DTI is sensitive to detect the white matter injury of TBI.FA value detected in the CC can help diagnose DAI earlier and evaluate the degree of injury.

Objective: To explore the change of bone mass density(BMD) and micronutrients in middle-aged and old patients with iron overload and the effect on levels of micronutrients.Methods: BMD was measured by dual energy X-ray absorptiometry(DXA) in 19 middle-aged and old patients with iron overload and 41 age-and gender-matched healthy subjects.Serum selenium(Se)was measured by fluorospectrophotometry.Serum zinc(Zn) was measured by inductively coupled plasma atomic emission spectrometry(ICP-AES).Plasma vitamin A,D and E were measured by HPLC.The levels of blood lipid and plasma homocysteine were measured by automatic biochemical analyzer.Results: Plasma vitamin E of 19 iron overload patients was significantly lower than control group,and the levels of triglyceride and homocysteine increased significantly(P

Objective To analyze the status of information release and the ability to identify public needs for hypertension prevention and control information in Shanxi and Hebei areas. Methods A systematic data collection method was used to obtain various information about hypertension prevention and control published in Shanxi and Hebei provinces. Based on the Delphi method, sensitive and evaluation indicators for the tertiary prevention of hypertension were determined. To evaluate the degree to which public needs were met in the two areas, the authority, timeliness, continuity, systematicness, and credibility of identifying public needs were calculated. Results The release of information on hypertension prevention and control in Shanxi and Hebei lagged behind that in World Health Organization (WHO) for 2 and 27 years, respectively, and this was mainly carried out by research institutes, with no response from professional associations and government agencies. Information on primary and secondary prevention was first released in 1980 in Shanxi province, while primary prevention information was first released in 2005 in Hebei province. Regarding hypertension prevention and treatment, Shanxi province could meet 6.11% of the public need, and Hebei province could meet 3.57% of it. The five dimensions of authority, timeliness, continuity, systematicness, and credibility of identifying the public need were 11.34%, 25.29%, 0, 50.31%, and 0 in Shanxi province, and 6.86%, 14.54%, 0, 50.00%, and 0 in Hebei province, respectively. Conclusions The ability to identify the public need for information on hypertension prevention and control in Shanxi and Hebei provinces is relatively low. Relevant departments should take measures to enhance the two‐way communication of information and improve the public's knowledge and participation.

Objective:To investigate the effects of disialyllacto-N-tetraose (DSLNT) on low molecular weight metabolic profile of intestinal contents in neonatal rats with necrotizing enterocolitis (NEC), in an attempt to explore the protective mechanism of DLSNT on intestinal tract of neonates.Methods:Immediately after birth, SD rats were randomly divided into the control group, the NEC group and the NEC+ DSLNT group according to random number tale method.All rats were hand-fed by special formula milk.Rats in the NEC group and NEC+ DSLNT group were exposed to hypoxia (950 mL/L nitrogen, 10 min, thrice per day) and cold stress (4 ℃, 10 min, thrice per day) for continuous 3 days to establish rodent NEC model.Rats in the NEC+ DSLNT group were hand-fed with special formula containing 300 μmol/L DSLNT.All rats were sacrificed after 72 h, and intestinal contents were collected from ileum and colon, followed by untargeted metabolomic determination with the ultrahigh-performance liquid chromatography Q extractive mass spectrometry (UHPLC-QE-MS) method.The terminal ileum was examined by hematoxylin-eosin staining.The metabolome data were analyzed with multivariable analysis using SIMCA 14.1.The metabolites that met both variable importance in the projection (VIP) >1 in the orthogonal partial least squares analysis (OPLS-DA) model and P<0.05 in the t-test were screened as differential metabolites between groups. Results:DSLNT reduced the incidence of NEC and pathological scores of ileum tissue from neonatal rats with NEC [3.0(2.0, 3.0) scores vs.1.0(1.0, 2.0) scores, P<0.01], and also significantly suppressed inflammatory infiltration.OPLS-DA model based on the metabolome data determined by UHPLC-QE-MS could perform effective discrimination between the NEC group and the control group, as well as the NEC+ DSLNT group and the NEC group.There were 64 differential metabolites between the NEC group and the control group (VIP value>1 and P<0.05 for the OPLS-DA model). These metabolites included docosahexaenoic acid (+ 288.0%, P=0.028), xanthine (+ 372.1%, P=0.007), L-arginine (+ 233.1%, P=0.027), L-leucine (+ 232.7%, P=0.015), N-acetylneuraminic acid (-41.6%, P=0.014), and so forth.These metabolites were associated with 34 metabolic pathways.Among them, such 6 pathways as arginine biosynthesis, arginine and proline metabolism were the most disturbed pathways affected by NEC.There were 15 diffe-rential metabolites in between NEC+ DSLNT group and NEC group, which included D-mannose (-73.5%, P=0.032), xanthine (-63.4%, P=0.008), linoleic acid (+ 137.9%, P=0.047), nicotinamide adenine dinucleotide (+ 278.2%, P=0.005), and so forth.These metabolites were mapped to 7 metabolic pathways, among them, linoleic acid metabolism pathway was the most relevant differential pathway affected by DSLNT.There were 8 overlapped meta-bolites in both comparison strategies, and the variation trend of these overlapped metabolites in the NEC group was significantly reversed by DSLNT supplementation. Conclusions:DSLNT could significantly attenuate the NEC pathological damage caused by hypoxia/cold stress in neonatal rats.This protective effect is associated with the improvement of the metabolic profile of intestinal contents caused by NEC and the modulation of the linoleic acid metabolic pathway.The early preventive supplementation of DSLNT is of great significance in maintaining neonatal intestinal homeostasis and preventing the process of NEC.

OBJECTIVETo develop and evaluate a mouse model of nonalcoholic steatohepatitis (NASH) induced by a high-fat and high-fructose (HFHFr) diet.

METHODSSix-week-old C3H mice were randomly divided into groups for HFHFr diet experimental modeling, high fat-only (HF) diet controls, high fructose-only (HFr) diet controls, and standard chow (SC) diet controls. The standard HFHFr diet was modified so that it consisted of 76.5% standard chow, 12% lard, 1% cholesterol, 5% egg yolk powder, 5% whole milk powder, and 0.5% sodium cholate, along with 20% fructose drinking water. At the end of experimental weeks 4, 8, and 16, measurements were taken for the NASH-related parameters of body mass, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), lipid profile, and wet liver weight (upon sacrifice). In addition, histological changes in the liver were evaluated by hematoxylin-eosin (HE) and oil red O staining. The significance of differences between groups was assessed by statistical analysis, using the

METHODSof t-test, Wilcoxon rank sum test, x2 test, F test or Fisher's test as appropriate.

RESULTSAs compared to the mice in the SC group at the corresponding time points, the mice in the HFHFr and HF groups showed significantly higher body mass and wet liver weight, as well as more extensive and robust lipid disposition in hepatic tissues as evidenced by oil red O staining. However, HE staining indicated that the HFHFr and HF groups had different degrees of macrosteatosis accompanied with intralobular inflammatory foci, with the former showing more remarkable NASH-related histological changes. Analysis at the end of week 16 showed that about 80% of the mice in the HFHFr group had developed NASH [nonalcoholic fatty liver disease (NAFLD) activity score (NAS): less than 5]. The levels of low-and high-density lipoprotein (LDL and HDL) cholesterol, as well as the levels of ALT and AST, were increased from the end of week 4 to the end of week 8 for the HFHFr and HF groups. At the end of week 16, the two groups differed in the extent of increase in total cholesterol and LDL and HDL cholesterol, with only the HFHFr group showing statistically significant changes. Specifically, at the end of week 16, the HFHFr group showed ALT levels of 108.5 +/- 93.34 U/L (F=5.099, P =0.005 vs. HF group: 44.30 +/- 35.71 U/L, HFr group: 46.70 +/- 17.95 U/L, SC group: 24.70 +/- 6.57 U/L), AST levels of 316.30 +/- 208.98 U/L (F=6.654, P=0.001 vs. HF: 132.12 +/- 75.43 U/L, HFr: 143.30 +/- 38.53 U/L, SC: 122.60 +/- 12.76 U/L), total cholesterol levels of 5.18 +/- 0.58 mmol/L (F=72: 470, P =0.000 vs. HF: 3.94 +/- 0.75 mmol/L, HFr: 2.30 +/- 0.50 mmol/L, SC: 2.02 +/- 0.24 mmol/L), HDL cholesterol levels of 3.05 +/- 0.49 mmol/L (F=25.413, P =0.000 vs. HF: 2.65 +/- 0.54 mmol/L HFr: 1.77 +/- 0.47 mmol/L, SC: 1.58 +/- 0.16 mmol/L), LDL cholesterol levels of 1.11 +/- 0.23 mmol/L (F =83.297, P =0.000 vs. HF: 0.72 +/- 0.17 mmol/L, HFr: 0.27 +/- 0.04 mmol/L, SC: 0.20 +/- 0.05 mmol/ L).

CONCLUSIONThe present study suggests that a mouse model of NASH can be successfully induced by a 16-week modified HFHFr diet.

Objective To explore the relationship of two loci (rs2293152, rs9579700) gene polymorphisms of signal transduction and transcription factor-3 (STAT3) with susceptibility to Henoch-Scho. .nlein purpura (HSP) and HSP nephritis (HSPN). Methods From September 2013 to September 2015, 180 children with HSP (group HSP) and 205 healthy children (control group) were enrolled in this study. Single nucleotide polymorphism (SNP) of intron 11 rs2293152C/G and intron 23 rs957970C/T of STAT3 gene was detected by sequence specific primer polymerase chain reaction (SSP-PCR). Results The frequency of CC genotype in STAT3 gene intron 11 rs2293152 in HSP group (26.1％) was significantly higher than that in control group (8.8％), and the frequency of allele gene of rs2293152C in HSP group (48.6％) was significantly higher than that in control group (32.7％) (P=0.013, 0.025). There were no differences in distribution of genotype and allele in rs957970C/T loci of intron 23 of STAT3 gene between two groups (P>0.05). The frequencies of genotype and allele of the two loci of STAT3 gene were no difference between HSPN and non HSPN groups (P>0.05). Conclusions The allele gene C of intron 11 rs2293152C/G of STAT3 gene may be a susceptible gene of HSP, while there was no association of 23 rs957970C/T polymorphism to HSP and there was no association of the two loci polymorphisms to HSPN.

Objective:To study the expression of zonula occludens-1(ZO-1) in neonates with necrotizing enterocolitis (NEC) and to explore the effects of disialyllacto-N-tetraose (DSLNT), a compound extracted from human milk, on the intestinal barriers in rat model of NEC.Methods:(1) Human study: From Feb 2013 to Dec 2020, the pathological samples of ileum tissue from 21 neonates (12 patients with NEC and 9 with intestinal atresia) from Pathology Department of our hospital were collected. The expressions of ZO-1 in these samples were examined using immunohistochemistry (IHC) method. (2) Animal study: A total of 28 newborn rats were randomly assigned into control group ( n=8), NEC group ( n=10) and DSLNT+NEC group ( n=10). Experimental NEC model was established based on hypoxia (95%N 2 10 min) /cold exposure (4 ℃ 10 min) three times a day for consecutive 3 days. DSLNT+NEC group were fed with formula+DLSNT (300 μmol/L) during hypoxia/cold exposure. All the surviving rats were sacrificed at the end of the experiment (72 h) and the terminal ileum tissues were collected. Hematoxylin-Eosin (HE) staining was used to evaluate tissue damage and Western blotting was used to determine the expressions of ZO-1. (3) Cellular study: Bacterial lipopolysaccharide (LPS) was used to establish a cellular inflammation model in human intestinal epithelial cell lines (Caco-2) and DSLNT (300 μmol/L) was applied to this model. Thiazolyl blue assay was used to examine cell viabilities and immunofluorescence assay was used to detect ZO-1 expression. The effects of DSLNT on cell growth and tight junctions of Caco-2 cells were analyzed. Results:(1)Human study: The villi of mucous layer of the lesion were damaged in NEC patients. ZO-1 expressions at the epithelial junction of NEC patients were decreased compared with intestinal atresia patients and non-lesion intestines of NEC patients. (2)Animal study: Apical extrusion, necrosis and shedding of epithelial cell were seen at the lesions in NEC group. The expression of ZO-1 in NEC group was significantly lower than the control group and DSNLT+NEC group ( P<0.05).DSNLT+NEC group had higher survival rates (8/10 vs. 6/10) and lower ileum inflammatory pathological scores [2.0(1.0, 2.8) vs. 3.5(3.0, 4.0)] than NEC group. (3) Cellular study: Caco-2 cells exposed to LPS showed inhibited cell growth and decreased ZO-1 immunofluorescence staining. Caco-2 cells in the DSLNT+LPS group showed better viability than LPS group and were comparable with the control group. The expression of ZO-1 was significantly increased in the DSLNT+LPS group. Conclusions:Tight junction injury of the intestinal epithelial cell is an important characteristic of NEC. ZO-1 is a potential target for the prevention and treatment of NEC. DSLNT may protect the neonatal intestines by modulating the expression of ZO-1 and keeping tight junction integrity.