BACKGROUND:Human umbilical cord mesenchymal stem cel s are considered as novel seed cel s in bone tissue engineering. Cryopreservation is an effective method for storing cel s for a long time.
OBJECTIVE:To explore whether umbilical cord mesenchymal stem cel s of cryopreservation could be induced to differentiated into osteoblasts.
METHODS:Mesenchymal stem cel s were isolated from the Wharton’s jel y of human umbilical cord tissue by the tissue explant adherent method. Morphology of primitive cel s was observed by inverted microscopy. Immunophenotypes and cel cycle of umbilical cord mesenchymal stem cel s were measured using flow cytometry. After frozen storage for 6 months, the second passage of umbilical cord mesenchymal stem cel s was thawed and subcultured to passage 12. Upon induction with osteogenic inductive medium, the osteogenic ability of passage 12 of umbilical cord mesenchymal stem cel was evaluated by alkaline phosphatase activity, the immunofluorescent analysis of osteocalcin and bone sialoprotein and the assay of alizarin red staining separately.
RESULTS AND CONCLUSION:Primary umbilical cord mesenchymal stem cel s displayed a typical fibroblast-like morphology. Flow cytometry showed that the cultured cel s expressed high levels of the mesenchymal stem cel s surface markers CD73, CD105 and CD90, but did not express hematopoietic cel s surface markers CD34 and CD45. The survival rate of umbilical cord mesenchymal stem cel s after resuscitation was 90%. The cel cycle analysis indicated that 75%of the cel s of passage 8 were in G 0/G 1 phase and 25%in S+G 2 M phase. Passage 12 cel s treated with osteogenic inductive medium displayed a higher alkaline phosphatase activity compared with control cel s (P<0.01). Moreover, the cel s, induced in osteogenic inductive medium, were positive for osteocalcin and bone sialoprotein staining and formed the mineralized nodules. Umbilical cord mesenchymal stem cel s stil maintain their biological characteristics after cryopreservation, and can be induced into osteoblasts with osteogenic inductive medium.

Objective To study the changes of portal vein pressure before and after interventional embolization with α-cyanoacrylate alkyl and their clinical significance. Methods Thirty five patients with esophageal-gastric variceal bleeding were included. Esophageal-gastric fundal varices were embolized with e-cyanoacrylate alkyl in 35 patients, and 10 of them were treated with combination of partial splenic emblization. Portal pressure was measured directly before and after interventional embolization. The patients were followed up for 3-18 months. The changes of the liver functions and complications after the embolization were observed. Results All bleeding were controlled successfully. The main portal pressure in those who received variceal embolization only increased significantly from (32.54±5.23) cm H2O (1 cm H2O=0. 098 kPa) to (37.45±5. 11) cm H2O; superior mesenteric vein pressure increased from (31.46±4.35) cmH2O to (34.33±4.68)cm H2O; and the level of serum albumin raised from (30.45±5.78) g/L to (34.57±6.84) g/L 3 weeks after embolization. Whereas the main portal pressure in those who received both variceal and partial splenic embolizations, decreased significantly from (32. 68±4. 89) cm H2O to (28. 70±4. 58) cm H2O; superior mesenteric vein pressure decreased from (31.46±4.35) cm H2O to (28.03±4.12) cm H2O; and splenic vein pressure decreased from (32.89±4.79) cm H2O to (28.81±5. 12) cm H2O.Conclusions Esophageal-gastric variceal embolization is effective for varieeal bleeding. The increase of main portal pressure after embolization may be benefit for liver function, while the rise of superior mesenteric vein pressure may increase the risk of portal hypertensive gastrointestinal re-bleeding.Esophageal-gastric variceal embolization combined with partial splenic embolization may reduce the incidence of portal hypertension and gastrointestinal re-bleeding, but will increase the risk of ascites.

Objective To investigate the clinical application of epinephrine hydrochloride in the prevention of bone cement implantation syndrome in the cemented hip replacement. Methods The clinical data of 48 patients treated with cemented hip replacement from July 2008 to April 2009 were retrospectively analyzed. All the patients were divided into control group and intervention group. The bone marrow cavities of 24 patients in the control group were not pretreated with saline epinephrine hydrochloride before implantation of bone cement; the bone marrow cavities of 24 patients in the intervention group were pretreated with saline epinephrine hydrochloride before implantation of bone cement. Systolic blood pressure (SBP), diastolic blood pressure (DBP), the mean arterial pressure (MAP), heart rate (HR)and pulse oxygen saturation ( SPO2 ) were compared between the two groups before bone cement implantation and 1,2, 3, 4, 5, 6, 7, 8, 9, 10 minutes after bone cement implantation. The data were analyzed with variance analysis and Q test. Results (1) In the control group: the blood pressure was decreased in control group one minute after bone cement implantation and a significant decrease of the blood pressure was observed at 2-6 minutes after the implantation ( P ＜ 0. 01 ). The blood pressure was increased seven minutes after the implantation, with the most significant increase of DBP ( P ＜ 0.05 ).The blood pressure recovered to normal 10 minutes later. The SPO2 was decreased significantly ( P ＜0.05 ) but no significant change was observed in HR ( P ＞ 0.05). (2) In the intervention group: the bone marrow cavity was pretreated with saline epinephrine hydrochloride before implantation of bone cement.ity. No significant difference was found in SBP, DBP, MAP, HR and SPO2 at different time points before and after bone cement implantation (P ＞0.05 ). Significant decrease of blood pressure and SPO2 was observed in control group and a significant hemodynamic change was detected at 2-6 minutes after the bone cement implantation. In the intervention group, no hemodynamic change was found in all the patients except that one patient was found with decrease of blood pressure and another one with the occasional premature ventricular contractions. Conclusion Pretreatment of bone marrow cavity with saline epinephrine hydrochloride can effectively prevent bone cement implantation syndrome.

OBJECTIVE:To investigate the compatibility stability of Fat-soluble vitamin (Ⅱ)/Water-soluble vitamin for in-jection with common electrolytes. METHODS:Referring to clinical common dose,Fat-soluble vitamin (Ⅱ) for injection/Wa-ter-soluble vitamin for injection collective packing [containing Fat-soluble vitamin(Ⅱ) for injection 2 ampoules and Water-soluble for injection 1 ampoule] were respectively mixed with Glucose injection,Potassium chloride injection,Concentrated sodium chlo-ride injection,Sodium bicarbonate injection,Potassium aspartate injection,Potassium aspartate and magnesium aspartate injection, Sodium glycerophosphate injection,Multi-trace elements injection (Ⅱ) to obtain mixture A-H. At room temperature (25 ℃),these mixtures were investigated in terms of appearance,pH value,osmotic pressure molar concentration and the number of insolu-ble particles at 0,1,2,3,4 h. The contents of bacterial endotoxin were tested at 0,4 h. RESULTS:Within 4 h after mixing, there was no significant change in appearance of those mixtures;pH value of mixture H changed greatly (RSD=5.13%,n=5), and that of mixture D and G increased significantly. The osmotic pressure molar concentration of those mixtures had no significantly change(RSD<2%,n=5)and lower than 600 mOsmol/kg. The bacterial endotoxin tests of those mixtures were negative. Two and four hours after mixing,the number of insoluble particles ≥10 μm in mixture B was increased significantly;2,3,4 h after mix-ing,the number of insoluble particles ≥10 μm in mixture E,F,H were increased significantly;0 ,1,2,3,4 h after mixing,the number of insoluble particles ≥10 μm in mixture G was increased significantly. There was statistical significance in the number of insoluble particle ≥10 μm in above mixtures compared to mixture A at the same time point(P<0.05),but it was in line with the standard of 2015 pharmacopeia. One,two,three and four hours after mixing,the number of insoluble particle ≥10 μm in mixture D was increased significantly,there was statistical significance compared to mixture A at the same time point(P<0.05);the num-ber of insoluble particle ≥10 μm in mixture D was beyond prescribed scope of pharmacopeia at 2,3,4 h after mixing. Within 4 h after mixing,both the number of insoluble particle ≥10 μm in mixture C and the number of insoluble particle ≥25 μm in edch mixture had no significant change,in accordance with pharmacopeia standard. CONCLUSIONS:Fat-soluble vitamin (Ⅱ)/Wa-ter-soluble vitamin for injection is not suitable for mixing with Multi-trace elements injection(Ⅱ),Sodium glycerophosphate injec-tion or Sodium bicarbonate injection due to great change of pH value and the number of insoluble particles.

A mixture of fructose and glucose was developed to simulate the hydrolysate of Jerusalem artichoke tubers, the fructose-based feedstock suitable for butanol production. With the initial pH of 5.5 without regulation during mixed-sugar fermentation, as high as 23.26 g/L sugars were remained unconverted, and butanol production of 5.51 g/L were obtained. Compared with either glucose or fructose fermentation, the early termination of mixed-sugar fermentation might be caused by toxic organic acids and the low pH. When the pH of the fermentation system was controlled at higher levels, it was found that sugars utilization was facilitated, but less butanol was produced due to the over-accumulation of organic acids. On the other hand, when the pH was controlled at lower levels, more sugars were remained unconverted, although butanol production was improved. Based on these experimental results, a stage-wise pH regulation strategy, e.g., controlling the pH of the fermentation system at 5.5 untill the OD620 reached 1.0, and then the pH control was removed, was developed, which significantly improved the fermentation performance of the system, with only 2.05 g/L sugars unconverted and 10.48 g/L butanol produced.
Acetone ; metabolism ; Butanols ; metabolism ; Fermentation ; Fructose ; metabolism ; Glucose ; metabolism ; Helianthus ; metabolism ; Hydrogen-Ion Concentration

Objective: To summarize the global research hotspots and trends of family intervention on autism spectrum disorder (ASD), to provide ideas for the development of localized research on family intervention for ASD in China. Methods: This study used the bibliometric software CiteSpace to conduct visual analysis on emerging cutting edge documents in the Web of Science core database in the field of family intervention for autism in the past 20 years (2001-2020). Data collection used 7 subject terms related to family caregiver intervention in February 2021. A total of 2 342 literature was obtained after data sorting. Results: The number of international publications has risen rapidly since 2013. The average annual number of international publications was 29 during 2001- 2012, then 250 during 2013-2020, which increased by 8 times. Among them, the United States ranked the top of publication with 1 295 articles, while publications from two universities in Canada and the United Kingdom showed higher impacts. The high frequency keywords that reflected the international hotspots included children, adolescent, mother, behavior, and stress; the high centrality keywords included follow up, intellectual disability, early recognition, young children and language. Conclusion The research hotspots in the field of ASD family intervention in the past 20 years included targets, strategies and forms, as well as the implementation effects of ASD family interventions. The trends of research in the past five years (2016-2020) mainly focus on the intervention studies of children, adolescents and adults with ASD and the randomized controlled trials of ASD family intervention focusing on caregivers.

Butanol production from acid hydrolysate of Jerusalem artichoke juice by Clostridium acetobutylicum L7 was investigated, and it was found that natural components of the hydrolysate were suitable for solvent production with the species. With batch fermentation using the medium containing 48.36 g/L total sugars, 8.67 g/L butanol was produced at 60 h, and the ratio of butanol to acetone to ethanol was 0.58:0.36:0.06, which were similar to the fermentation with fructose as carbon source, but both of these two fermentations were slower than that with glucose as carbon source, indicating the fructose transport of the species might not be effective as that for glucose. When the total sugars of the medium were increased to 62.87 g/L, the residual sugars increased slightly from 3.09 g/L to 3.26 g/L, but butanol production of the fermentation system was improved significantly, with 11.21 g/L butanol produced and the ratio of butanol to acetone to ethanol at 0.64:0.29:0.05, which illustrated that an excess in sugars enhanced the butanol biosynthesis of the species by compromising its acetone production. When the sugar concentration of the medium was further increased, much more sugars were remained unconsumed, making the process economically unfavourable.
Butanols ; metabolism ; Clostridium acetobutylicum ; metabolism ; Fermentation ; Helianthus ; chemistry ; Industrial Microbiology ; methods

Objective:To explore the role and mechanism of kidney brain protein (KIBRA) down-regulation in cognitive dysfunction caused by chronic cerebral hypoperfusion.Methods:Ninety male SPF grade Sprague Dawley (SD) rats were divided into four groups according to random number table: sham operation group ( n=15), chronic hypoperfusion group (2VO group, n=25), chronic hypoperfusion stereotaxic injection of AAV-KIBRA group (2VO+ AAV-KIBRA group, n=25), chronic hypoperfusion stereotaxic injection of AAV-Vector group (2VO+ AAV-vector group, n=25). Chronic cerebral hypoperfusion model was established by bilateral ligation of common carotid artery, and stereotactic injection of 2 μL AAV-KIBRA or AAV-vector was performed for 30 days.Morris water maze, in vitro electrophysiology, p21-activated kinase 3(PAK3) activity detection, Western blot, immunoprecipitation and Golgi staining were used to detect spatial learning and memory ability, long-term potentiation(LTP), KIBRA level expression, PAK3 activity changes and the distribution of dendritic spines.SPSS 16.0 statistical software was used for statistical data.One-way ANOVA was used to compare the differences between groups.LSD test was used to compare the significance of data differences between the two groups.Welch test was used for uneven variance. Results:After 1 month of chronic cerebral hypoperfusion, the level of KIBRA in the hippocampus of rats was detected by homogenate and Western blot, and it was found that the level of KIBRA in 2VO group was lower than that of sham group(73.49±4.12)% ( P<0.01). AAV-KIBRA injection in hippocampal CA1 region significantly up-regulated the level of KIBRA to (91.91±7.01)% over 2VO group ( P<0.01). Morris water maze test showed that the latency of the 2VO group(3rd-7th day trail data: (48.18±2.82)s, (43.45±2.27)s, (32.27±2.22)s, (26.55±2.37)s, (17.18±2.67)s) were significantly longer than those of the sham group((41.67±2.74)s, (32.58±2.57)s, (22.50±2.94)s, (16.91±2.39)s, (8.75±1.52)s) (all P<0.05), and the latencies of the 2VO+ AAV-KIBRA group 3rd-7th day trail data: (43.83±2.95)s, (35.25±2.15)s, (26.58±2.03)s, (19.92±2.17)s, (17.75±1.35)s) was significantly shorter than that of the 2VO group ((all P<0.01). The Morris water maze test with the platform removed showed that the latency of rats in the 2VO group to reach the platform region was significantly longer than that of the sham group, while the latency of rats in the 2VO+ AAV-KIBRA group to reach the platform region was significantly shorter than that in the 2VO group ( P<0.01). At the same time, the retention time and the crossing times in the platform region of 2VO group were less than those of the sham group ( P<0.01), but the retention time and the crossing times in the platform region of 2VO+ AAV-KIBRA group were significantly higher than those in the 2VO group ( P<0.01). The electrophysiological records of the brain slices showed that the relative excitatory postsynaptic field potential of 2VO group (1.43±7.43) was significantly lower than that of sham group (2.21±6.54) after high frequency stimulation, while the relative excitatory postsynaptic field potential of 2VO+ AAV-KIBRA group (1.90±8.15) was higher than that of 2VO group ( P<0.01). Immunoprecipitation in rat hippocampus revealed that PAK3 could be detected by Western blot assay when KIBRA was precipitated.The results showed that the relative enzyme activity of PAK3 in 2VO hippocampal tissue (0.64±0.04) was significantly lower than that in sham group (1.02±0.07), while the relative enzyme activity of PAK3 in 2VO+ AAV-KIBRA group (0.86±0.03) was significantly higher than that in 2VO group.Golgi staining showed that the density of dendritic spines in 2VO hippocampal neurons((6.85±0.43)/10 μm) was significantly lower than that in sham group((11.83±0.58)/10 μm), while the density of dendritic spines in 2VO+ AAV-KIBRA group((10.22±0.39)/10 μm) was significantly higher than that in 2VO group. Conclusion:The down-regulated of KIBRA after chronic cerebral hypoperfusion plays a key role in cognitive dysfunction and is also involved in the decrease of synaptic functional plasticity.The downregulation of KIBRA is involved in the structural plasticity of dendrites through the regulation of PAK3 activity.Therefore, KIBRA may be an important target for the prevention and treatment of cognitive function of chronic cerebral hypoperfusion.

Objective: To identify differentially expressed genes in the transcriptome of the lesional versus nonlesional skin tissues of patients with moderate and severe atopic dermatitis (AD) , and to elucidate their roles in the pathogenesis of AD. Methods: From July to October in 2016, lesional and nonlesional skin tissues were obtained from 5 outpatients of Han nationality with AD in Guangzhou Institute of Dermatology, Institute of Dermatology, Guangzhou Medical University. The next-generation high-throughput transcriptome-wide RNA sequencing (RNA-seq) was performed to identify differentially expressed genes, which were subjected to GO function annotation and KEGG pathway analysis. Real-time fluorescence-based quantitative PCR (qRT-PCR) was conducted to verify differences in candidate gene expression between lesional and nonlesional skin tissues. Results: An average of 10.96 GBs sequence reads were acquired among 10 samples. A total of 21 729 genes were detected, including 19 268 known genes and 2 545 predicted novel genes. A total of 23 153 new transcripts were detected, of which 18 889 were new alternative splicing subtypes of known protein-coding genes, 2 545 were transcripts belonging to new protein-coding genes, and the remaining 1 719 belonged to long-stranded non-coding RNA. Totally, 78 differentially expressed genes were identified between the lesional and nonlesional skin tissues, including 69 upregulated and 11 downregulated genes in the lesional skin tissues. Among them, there were several genes known to be associated with AD inflammation (CXCL1/2/8, IL6/IL1β, MMP1, SERPINB4, S100A2, GZMB, OASL, OSM) and barrier (KRT16, FABP5, CYP1A1) and keratinocyte differentiation (IL-20) . GO analysis revealed that functions of 72 differentially expressed genes could be annotated. KEGG pathway analysis showed that the differentially expressed genes were grouped into 132 signaling pathways, of which 13 were significantly enriched, including the interleukin (IL) -17 pathway, NOD-like receptor signaling pathway, Toll-like receptor signaling pathway, etc. qRT-PCR showed that the mRNA expression levels of candidate genes CXCL1, KRT6A, IL36A, SERPINB4 and PSAPL1 was consistent with the transcriptome sequencing results. Conclusions Differentially expressed genes and related important regulatory signaling pathways were identified between the lesional and nonlesional skin tissues of patients with AD at the transcriptional level, and the IL-17 pathway was found to be mostly enriched in AD lesions in patients of Han nationality. These findings provide an important basis for further study on the pathogenesis of AD..

Objective To understand the various HPV types and the factors influencing their distribution among women with normal cervical cytology in the Shanghai area, to provide basic data for cervical cancer prevention and vaccine use.Methods A questionnaire-based survey was conducted among 3 372 married women in the Shanghai area. A PCR reverse dot blot (PCR-RDB) method was adopted for HPV genotyping of cervical exfoliated cell samples from 3 206 women. Odds ratio (OR) for HPV infection were analyzed using logistic regression. Results Of the 3 206 women investigated, 669 (20.87%) were positive for HPV infection. The highest incidence of HPV infections was seen in the age groups of 55-59 years, 50-54 years and 45-49 years, with the rates of positive detection being 27.67%, 21.65%, and 21.55%, respectively. While 70.4% of the positive cases had a single infection, 29.6% showed multiple infections. In cases with multiple infections, double infection was predominant (20.63%). The top five high-risk gene types were HPV52(3.65%),HPV53(2.71%),HPV51(2.03%),HPV58(1.87%),and HPV16 (1.40%). The top three low-risk gene types were HPV81 (2.03%), HPV42 (1.43%), and HPV55 (1.31%). Among the women with HPV infections,there were 483 having medium-high-risk HPV infection.There were significant differences in the age,the age at first sexual activity,sexual activities per week,education,and alcohol consumption between HPV positive and HPV negative patients. Age stratification showed that the infection rates in the 55-59 years old group were significantly higher than that in the other age groups(χ2=15.349, P=0.000). Both single factor and multivariate non-conditional logistic regression analyses showed that higher education and the start of sexual activity at a later age were protective factors for medium-high-risk HPV infection,with regression coefficients of-0.165 and-0.08,respectively (P<0.01) in the multivariate analysis. The risk factors included age between 55- 59 years, menstrual status (menopause), sexual activity (≥3 times per week) and alcohol consumption. High-risk HPV infections also had the same risk factors,and the odds ratios were 1.558,1.275,and 1.678,respectively(P<0.01).However, 55-59 years of age and alcohol consumption are independent risk factors for medium-high risk HPV infection. Conclusions High-risk HPV in women of Shanghai is commonly caused by HPV52, HPV53, HPV51,HPV58,and HPV16.The high-risk group includes women who are 55-59 years old.While drinking is an independent risk factor for medium-high risk HPV infection, a moderate sex life and delayed age at first sex can reduce the risk of high-risk HPV infections.