Aim To explore the effects of astragaloside(AS)on the proliferation of neural stem cells(NSCs).Methods The NSCs of embryonic day(E)16 SD rat were cultured in vitro.The NSCs were identified by the immunocytochemical(ICC)staining of Nestin.The ICC staining of BrdU was adopted to characterize the proliferation of NSCs.The number of neurospheres and the ICC staining of BrdU were performed to identify their proliferation properties.Real time RT-PCR technique was used to investigate the proliferation mechanisms of the AS on the NSCs.Results Characteristic protein(Nestin)of NSCs labeling,BrdU labeling were positive showed by immunocytochemical staining.The ICC of BrdU labeling results showed that different dosages of AS could promote the proliferation of NSCs in vitro.The proliferation rate of NSCs was increased extremely significant(P

OBJECTIVE:To evaluate the rationality and validity of chemical medicine tablet production equipment cleaning pro-cedure. METHODS:Among several chemical medicines prepared by similar production technology as Metoprolol succinate sus-tained-release tablets,Captopril tablets,Isosorbide mononitrate tablet and Metformin hydrochloride tablet,Metoprolol succinate sus-tained-release tablets had strongest toxicity and were included in validation test. The production equipment was cleaned and disinfect-ed according to cleaning procedure. The point which was most difficult to clean could be wiped and sampled by using the cotton swab method. The detection limit and the limit of quantitation of the residue limits were verified as well as the recovery rate of wip-ing,in order to evaluate whether the results meet the requirements. RESULTS:The cotton swab method is adopted to wipe sample and detect the point which is most difficult to clean. The visible foreign body has not been found in each sampling point. The amount of residual drug is <29.75 μg/cotton bud,and microbial limits are <50 CFU/cotton bud,indicating test items are in line with the standard. CONCLUSIONS:The cleaning method can effectively clean the production equipment,and can effectively pre-vent product contamination and cross contamination to ensure the quality,efficacy and safety of the next batch of products.

OBJECTIVETo investigate the effect of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on the stability of resin-dentin bonds against pH cycling.

METHODSResin-bonded dentin specimens were prepared following manufacturers' instructions, and randomly divided into 3 groups. Among them, 2 groups experienced pH cycling, in which specimens applied CPP-ACP or distilled and deionized water (DDW) on the bonding interface, respectively. Microtensile bond strength (muTBS) testing, failure mode analysis, micromorphological and nanoleakage evaluation of bonding interface and elemental analysis within hybrid layer were performed after 15 days pH cycling. The other group was tested immediately after specimens' preparation without pH cycling.

RESULTSNo significant differences were found in muTBS between no pH cycling and pH cycling/CPP-ACP group. Their muTBS were both significantly higher than that of pH cycling/DDW group (P < 0.05). Mixed fractures were the most prevalent failure mode. The quality of hybrid layer in pH cycling/CPP-ACP group was better than that of pH cycling/DDW group, and the nanoleakage was also less severe. Comparing with pH cycling/DDW group, the atomic percentages of Ca in the other two groups were both significantly higher, while those of Ag were statistically lower (P < 0.05).

CONCLUSIONLocal application of CPP-ACP can promote the stability of resin-dentin bonding interface against pH cycling and prolong bonding degeneration.

Calcium Phosphates ; Caseins ; Dental Bonding ; Dentin ; Dentin-Bonding Agents ; Humans ; Phosphopeptides ; Resin Cements

Objective:To compare the effects of different lung recruitment maneuvers in infants undergoing laparoscopic surgery.Methods:A total of 70 pediatric patients of either sex, aged 1-6 yr, weighing 10-24 kg, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, scheduled for elective laparoscopic surgery from September 2020 to June 2021 with expected operation time≤2 h, were divided into 2 groups ( n=35 each) by a random number table method: recruitment maneuver using incremental positive end-expiratory pressure (PEEP) group (PV group) and recruitment maneuver using controlled lung expansion group (RM group). The children underwent pressure-controlled ventilation after tracheal intubation, and lung recruitment was performed at 20 min after pneumoperitoneum, immediately after pneumoperitoneum, and at the end of operation and before tracheal extubation.In PV group, PEEP was gradually increased, the upper limit of airway pressure was 35 mmHg, PEEP was increased by 5 cmH 2O, ventilation was performed for 30 s, then PEEP was increased to 15 cmH 2O, ventilation was continued for 30 s, then the parameters were adjusted to the original ones, and ventilation was continued until the next lung recruitment.In RM group, manual ventilation mode was used, the pressure valve was adjusted to 30 cmH 2O, the pressure was increased to the maximum by rapid oxygenation, the breathing cuff was manually squeezed until the airway pressure achieved 30-35 mmHg, and 30 s later ventilation was performed with the original ventilation parameters, lasting for 30 s until the next lung recruitment.Peak airway pressure and mean airway pressure were recorded at 5 min after tracheal intubation (T 1), 20 min after pneumoperitoneum (T 2), immediately after pneumoperitoneum (T 3) and before extubation after surgery (T 4), and dynamic lung compliance was calculated.Blood gas analysis was performed at T 2 and T 4, and arterial partial pressure of oxygen and arterial partial pressure of carbon dioxide were recorded, oxygenation index, alveolar-arterial oxygen partial pressure difference and respiratory index were calculated.Lung ultrasonography scores were assessed before tracheal extubation (T 0) and at T 4 and 20 min after entering the postanesthesia care unit (T 5). The time of tracheal extubation and length of postoperative hospital stay were recorded.Hypoxemia in postanesthesia care unit and occurrence of pulmonary complications within 3 days after operation were recorded. Results:Compared with RM group, peak airway pressure and mean airway pressure were significantly decreased at T 2, 3, dynamic lung compliance was increased at T 2-4, arterial partial pressure of oxygen and oxygenation index were decreased , arterial partial pressure of carbon dioxide, alveolar-arterial oxygen partial pressure difference and respiratory index were increased at T 2 and T 4, lung ultrasonography scores were decreased at T 4 and T 5, and the incidence of postoperative hypoxemia was increased, and tracheal extubation time was prolonged in RM group ( P<0.05). Conclusions:Lung recruitment maneuver using incremental PEEP provides better efficacy than that using controlled lung expansion in infants undergoing laparoscopic surgery.

Objective    To explore the effects of PKD1 gene on mouse aortic smooth muscle (MOVAS) cells autophagy. Methods    The shRNA and over-expression lentiviral vectors for the target gene of PKD1 were constructed. MOVAS cells were infected by a number of successful packaging shRNA (PKD1 knockdown) or ETS-1 (PKD1 over-expressing) lentiviral vectors, and qPCR was used to test interference and over-expressing effects. Then qPCR and Western blotting were used to detect the expression levels of autophagy markers including Atg5, Beclin1 and LC3 in control group, shPKD1 group and ETS-1 group. Results    Compared with the control group, PKD1 mRNA level was decreased in the shPKD1 group (P<0.05); ETS-1 and PKD1 mRNA levels were increased in the ETS-1 group (P<0.05). In contrast with the control group, the mRNA levels of autophagy markers including Atg5 (P<0.05) and Beclin1 (P<0.01) were obviously decreased in the shPKD1 group, but they were obviously increased in the ETS-1 group (P<0.001). Protein levels of Atg5, Beclin1 and LC3 were significantly decreased in the shPKD1 group (P<0.05), but they were increased obviously in the ETS-1 group (P<0.05) in contrast with the control group. Conclusion    PKD1 gene is involved in MOVAS cells autophagy, low expression of PKD1 gene can inhibit autophagy and high expression of PKD1 promotes autophagy in vascular smooth muscle cells.

OBJECTIVETo explore the immuno-effect of plasmacytoid dendritic cells (pDC) on bacteria infection induced spontaneous remission (SR) of leukemia.

METHODSBoth pDC and myeloid dendritic cells (mDC) were isolated and purified from leukemic patient with SR and healthy donor by combination of immunomagnetic beads and flow cytometry. pDC were cultured in RPMI1640 medium and stimulated with different bacteria. The T cells proliferation was detected by MTT, and cytokine production by ELISA kits.

RESULTSThe human bacterial pathogen Staphylococcus aureus and Pseudomonas aeruginosa stimulation for 48 h resulted in the maturation of pDC with production of high quantity of IFN-α at (15.34 ± 2.91) ng/ml and (10.38 ± 1.41) ng/ml, respectively, comparing with that of negative group at (1.36 ± 0.13) ng/ml (P<0.01). Activated pDC could promote the differentiation of naive CD4⁺ T cells to Th1 cells with secretion of IFN-γ at (2.16 ± 0.37) ng/ml and (2.73 ± 1.11) ng/ml, respectively, comparing with that of positive control at (2.55 ± 0.23) ng/ml (P > 0.05). Activated pDC showed higher T cell stimulatory capacities [proliferation index (PI) was 4.36 and 4.05, respectively] than that of non-activated pDC (PI was 1.23 and 0.13, respectively) (P < 0.01).

CONCLUSIONStaphylococcus aureus and Pseudomonas aeruginosa activated pDC may play a key role in SR of leukemia following severe infections.

CD4-Positive T-Lymphocytes ; Dendritic Cells ; immunology ; Flow Cytometry ; Humans ; Interferon-alpha ; Leukemia ; diagnosis ; immunology ; Lymphocyte Activation ; Pseudomonas aeruginosa ; immunology ; Remission, Spontaneous ; Staphylococcus aureus ; immunology

Clostridia inhabiting in jiupei and pit mud plays key roles in the formation of flavour during the fermentation process of Luzhou-flavour baijiu. However, the differences of Clostridial communities between jiupei and pit mud remains unclear. Here, the species assembly, succession, and metabolic capacity of Clostridial communities between jiupei and pit mud were analysed by high-throughput sequencing and pure culture approaches. The ratio of Clostridial biomass to bacterial biomass in the pit mud was relatively stable (71.5%-91.2%) throughout the fermentation process. However, it varied widely in jiupei (0.9%-36.5%). The dominant Clostridial bacteria in jiupei were Clostridium (19.9%), Sedimentibacter (8.8%), and Hydrogenispora (7.2%), while Hydrogenispora (57.2%), Sedimentibacter (5.4%), and Caproiciproducens (4.9%) dominated in the Clostridial communities in pit mud. The structures of Clostridial community in pit mud and jiupei were significantly different (P=0.001) throughout fermentation. Isolated Clostridial strains showed different metabolic capacities of volatile fatty acids in pure culture. Spatial and temporal heterogeneity of Clostridial communities existed in the baijiu fermentation pit, which was closely related to the main flavour components of Luzhou-flavour baijiu.
Alcoholic Beverages ; microbiology ; Bacteria ; classification ; metabolism ; Clostridium ; physiology ; Fatty Acids, Volatile ; metabolism ; Fermentation ; Food Microbiology

Multi-species solid-state fermentation in a mud pit is one of the typical features of strong-flavor baijiu, in which archaea plays important roles, however, the archaeal community distribution and diversity during fermentation are still lack of research. The biomass, composition and succession of archaea communities in fermented grains and pit mud were analyzed by high throughput sequencing. The potential interaction between archaea and bacteria was analyzed by co-occurrence network. Results demonstrate that the average biomass of archaea in pit mud was about 200 times higher than that of fermented grains. There was no significant difference in archaeal community structure between fermented grains and pit mud (r=0.017, P=0.074), but succession patterns between them showed significant correlation (r=0.30, P=0.03). Methanobacterium was the most abundant archaea in fermented grains and pit mud, and other dominant groups included Methanosarcina, Methanocorpusculum, Methanoculleus, and Methanobrevibacter. The co-occurrence network analysis showed that Methanobacterium was positively correlated with most bacteria in fermented grains and pit mud, especially with Hydrogenispora and Caproiciproducens, the dominant bacteria in pit mud. Our results revealed the temporal and spatial distribution characteristics and potential functions of the archaeal community in the mud pit of strong-flavor baijiu.
Alcoholic Beverages/analysis* ; Archaea/genetics* ; Bacteria ; Fermentation ; Taste

Objective To explore the effect of knocking down Rho-associated coiled-coil kinase (ROCK2) gene on the cognitive function of amyloid precursor protein/presenilin-1 (APP/PS1) double transgenic mice and its mechanism. Methods APP/PS1 double transgenic mice were randomly divided into AD model group (AD group), ROCK2 gene knock-down group (shROCK2 group), ROCK2 gene knock-down control group (shNCgroup), and wild-type C57BL/6 mice of the same age served as the wild-type control (WT group). Morris water maze and Y maze were employed to test the cognitive function of mice. Neuron morphology was detected by Nissl staining. Immunofluorescence histochemical staining was used to detect the expression of phosphorylated dynamin-related protein 1 (p-Drp1) and mitochondrial fusion 1 (Mfn1). Western blot analysis was used to detect the expression ROCK2, cleaved-caspase-3 (c-caspase-3), B-cell lymphoma 2 (Bcl2), Bcl2-related protein X (BAX), p-Drp1, mitochondrial fission 1 (Fis1), optic atrophy 1 (OPA1), Mfn1 and Mfn2. Results Compared with AD group mice, the expression of ROCK2 in shROCK2 group mice was significantly reduced; the cognitive function was significantly improved with the number of neurons in the hippocampal CA3 and DG areas increasing, and nissl bodies were deeply stained; the expression of c-caspase-3 and BAX was decreased, while the expression of Bcl2 was increased; the expression of mitochondrial division related proteins p-Drp1 and Fis1 were decreased, while the expression of mitochondrial fusion-related proteins OPA1, Mfn1 and Mfn2 were increased. Conclusion Knock-down of ROCK2 gene can significantly improve the cognitive function and inhibit the apoptosis of nerve cells of APP/PS1 mice. The mechanism may be related to promoting mitochondrial fusion and inhibiting its division.
Animals ; Mice ; Alzheimer Disease/pathology* ; Amyloid beta-Peptides/metabolism* ; Amyloid beta-Protein Precursor ; Apoptosis/genetics* ; bcl-2-Associated X Protein ; Caspase 3 ; Cognition ; Disease Models, Animal ; Mice, Inbred C57BL ; Mice, Transgenic ; Mitochondrial Dynamics/genetics*