Berberine could reverse the phenylephrine-induced contraction of aortic strips of rabbits, but not those induced by histamine, serotonin and high-K. It showed no effect on the resting tension of the strips.Prazosin l ?M completely abolished the phenylephrine-induced contraction of the strips, but berberine 100 ?M could block 94 ? 9%.The IC50 of berberine and prazosin for phenylephrine-induced c-ontraction is 30 ?M and 0.2 ?M. The results suggest that the action of berberine is not similar to sodium nitroprusside, but is similar to prazosin, a selective ?1 adrenoceptor blocker, although its potency is less than prazosin.

According to the onset age, a total of 5 674 outpatients with gout admitted in the affiliated hospital of Qingdao University from January 2011 to January 2016 were divided into youth group (≤44 years, n=3 058), middle age group (45~64 years, n=2 101), and elderly group (≥65 years, n=515).Their clinical and biochemical characteristics were analyzed.The results showed that the proportion of gout in the males in three groups was higher than that in the females.The proportions of gout in male youth, middle age, and elderly groups were 98.1%, 93.4%, and 83.1%, respectively.The proportion of gout in females increased with age.The proportion of gout family history in youth group was higher than that in middle age and elderly groups(P<0.05).The proportions of hypertension and impaired fasting glucose(IFG) in elderly group were higher than those in youth and middle age groups (P<0.05), with lower proportions of hypertriglyceride and obesity (P<0.05).The levels of systolic blood pressure, blood urea nitrogen, creatinine in elderly group were increased compared with youth and middle age groups (P<0.05) while the levels of uric acid and triglyceride were decreased (P<0.05), with higher ratio of renal dysfunction (P<0.05).There were no differences in tophus and kidney calculi among three groups (P>0.05).The distributions of the onset joint among three groups revealed statistical difference(P<0.05), but the first plantar toe joint was mostly involved in each group.These results suggest that the clinical characteristics of patients with gout at different onset ages are not identical, should be treated differently.

Objective: To investigate the factors affecting the development of non-proliferative diabetic retinopathy (NPDR) among patients with type 2 diabetic mellitus (T2DM), so as to provide insights into manangement of NPDR. Methods: T2DM patients without obvious eye discomfort at ages of 18 years and older admitted to Department of Endocrinology, Jining No. 1 People's Hospital during the period from December 2019 to May 2021 were enrolled. Participants' demographics, smoking, alcohol consumption, medical history of diabetes and use of medicines were collected, and the height, weight and blood pressure were measured. The levels of glycosylated hemoglobin (HbA1c), fasting blood glucose, blood C-peptide, lipid and creatinine were tested, and retinopathy was examined with a non-mydriatic fundus camera. The factors affecting the development of NPDR were identified among T2DM patients using a multivariable logistic regression model. Results: A total of 486 T2DM patients were enrolled, including 354 men (72.84%), with a median age of 48.00 (15.25) years, and median diabetes duration of 35.00 (104.25) months. The prevalence of NPDR was 19.34% among the participants. multivariable logistic regression analysis identified an educational level of senior high school and above (OR=0.546, 95%CI: 0.325-0.918), duration of diabetes (OR=1.008, 95%CI: 1.005-1.012), HbA1c (OR=1.183, 95%CI: 1.034-1.354) and use of non-sulfonylurea insulin secretagogues (OR=1.859, 95%CI: 1.082-3.196) as factors affecting the risk of NPDR among T2DM patients. Conclusion A high risk of NPDR is found among T2DM patients with a low educational level, long duration of diabetes, poor HbA1c control and use of non-sulfonylurea insulin secretagogues.

Objective To study the change characteristics of erythrocyte parameters in carriers of deletional alpha-thalassemia gene.Methods 389 patients with deletional alpha-thalassemia gene determined by the gap-PCR technique were classified into three groups based on different genotypes of alpha-thalassemia including silent thalassemia group,alpha-thalassemia trait group (265 ca-ses)and intermediate thalassemia group,and contemporaneous 188 healthy adults were randomly selected as the normal control-group(NC).The erythrocyte parameters including RBC,Hb,MCV,MCH,RDW were retrospectively analyzed and their differences were compared among aboved-mentioned groups by the analysis of variance and the multiple comparison.Results Alpha-thalasse-mia manifested by different degrees of microcyte hypochromia.There were statistically significant differences in the erythrocyte pa-rameters among various genotypes and phenotypes about erythrocyte indices(P <0.05).Moreover,the Hb,MCV and MCH values were lower than those in the control group,and had the decreasing tendency with the increase of deletedα-globin gene numbers;but the RDW value was higher than that in the control group,showing the increasing tendency,the differences had statistical siginifi-cance(P <0.05).Conclusion With the increase of deleted α-globin gene number,the characteristic of small-cell low-hemoglobin is more and more remarkable.RBC has a overall increase,while Hb,MCV and MCH have a decreasing tendency,and the heterogeneity of erythrocyte volume is increased.When MCV and MCH decreasing in high risk region,thalassemia should be highly suspected, but normal MCV and MCH can not exclude silent thalassemia and alpha-thalassemia trait.

HBV is a major health problem faced by human beings. The study of the mechanism of HBV infection has been a key point in this field. To carry out the related research on HBV, establishment of simple and available in vivo and in vitro infection models is the trend. Due to the close relationship between tree shrews and human/primates, the tree shrew model of HBV infection has attracted more and more attention. This review summarizes recent research, both at home and abroad, about in vivo and in vitro HBV infection in tree shrews, especially the research progress in in vitro culture of primary hepatocytes of tree shrews, and looks to the future research directions as well.

Objective:To investigate the application value of KARL iterative algorithm combined with low-dose CT scanning in young and middle-aged patients with femoral neck fractures.Methods:The clinical data of 132 young and middle-aged patients with femoral neck fractures diagnosed by clinical and CT scan in Taizhou Bo′ai Hospital from August 2019 to May 2022 were collected. There were 68 cases in the conventional dose group reconstructed by projection, and 64 cases in the low dose group reconstructed by KARL iterative algorithm. The subjective image quality score, the excellent and good rates and CT value, signal to noise ratio (SNR) were compared between the two groups. The effective dose (ED), CT dose index (CTDIvol), dose-length product (DLP) were compared between the two groups.Results:There were no significant differences in subjective image quality score, the excellent and good rates between the two groups ( P>0.05). There were no significant differences in CT values and SNR of the trabecular dense area at the central level of the femoral head, the iliopsoas muscle at the same level, and the anterior abdominal wall or buttocks subcutaneous fat at the same level between the two groups ( P>0.05). The levels of CTDIvol, DLP and ED in he low-dose group were lower than those in the conventional dose group: (3.35 ± 1.05) mGy vs. (12.90 ± 2.92) mGy, (66.33 ± 20.26) mGy/cm vs. (253.12 ± 58.57) mGy/cm, (0.99 ± 0.30) mSv vs. (3.79 ± 0.88) mSv, there were statistical differences ( P<0.05). Conclusions:Based on KARL iterative algorithm combined with low-dose CT scanning has no significant effect on the clinical diagnosis and classification of femoral neck fractures, the scanning image has a high objective evaluation value, and can effectively reduce the radiation dose received by patients, which is useful for clinical preoperative diagnosis and surgical planning.

Diabetic nephropathy (DN) is the leading cause of end-stage failure of the kidney, but the efficacy of currently available strategies for the prevention of DN remains unsatisfactory. In this study, we investigated the effects of free anthraquinones (FARs) extract, which was extracted from the rhubarb and purified by macroporous resin DM130 with gradient mixtures of ethanol/water as the lelution solvents, in high glucose-cultured glomerular mesangial cells (MCs). The cell proliferation was determined by CCK-8 assay, the levels of TGF-beta1, CTGF, ColIV and FN proteins in the supernatant of MCs were measured by ELISA assays, and the mRNA levels of these four genes were detected by RT-PCR. The results showed that the increased proliferation of MCs, the mRNA levels and protein expression of TGF-beta1, CTGF, ColIV and FN induced by high glucose were inhibited after the treatment with the FARs extract. This indicated that FARs extract could inhibit cell proliferation and the expression of main extracellular matrix induced by high glucose in MCs. The FARs extract exhibited potential values for prophylaxis and therapy of DN.
Anthraquinones* ; Cell Proliferation* ; Diabetic Nephropathies ; Enzyme-Linked Immunosorbent Assay ; Extracellular Matrix* ; Glucose* ; Kidney ; Mesangial Cells ; Rheum* ; RNA, Messenger ; Sincalide ; Solvents ; Transforming Growth Factor beta1

OBJECTIVE: To study the cytotoxicity of recombinate toxin MSH-Ang to Hep-2. METHOD: The depurated MSH-Ang were applied in cytotoxicity experiment, and the growth inhibiting action to laryngeal carcinoma cell Hep-2 were observed. RESULT: Recombination protein inhibited the growth of laryngeal carcinoma cell Hep-2, and its inhibiting action enhanced and corpuscular mortality rate increased along with the concentration increasing. CONCLUSION Recombinant toxin MSH-Ang can not only take special effect in tumors with high MSHR, but also target to many other popular tumors.
Angiopoietins ; genetics ; pharmacology ; Cell Line, Tumor ; Genetic Engineering ; Humans ; Laryngeal Neoplasms ; Melanocyte-Stimulating Hormones ; genetics ; pharmacology ; Recombination, Genetic

Objective:To investigate the resistance and transmission mechanism of carbapenem-resistant Enterobacterales (CRE), so as to provide the scientific evidence for the treatment and prevention of CRE infection.Methods:Seventy-six isolates of CRE isolated from Shaoxing Second Hospital between May 2016 and August 2018 were included. The isolates were re-identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The minimum inhibitory concentrations (MICs) of colistin, tigecycline, ceftazidime-avibactam, fosfomycin and other antibacterial drugs were determined using broth microdilution or agar dilution methods. PCR and sequencing analysis were performed to detect carbapenemase encoding genes ( blaKPC, blaNDM, blaIMP, blaVIM, blaOXA-48). Pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used to analyze homology of strains. S1-PFGE combined with Southern blot hybridization were used to locate the carbapenamase genes. Filter mating test were performed to determine the horizontal transfer ability of plasmids harboring carbapenamase genes. Results:Among the 76 isolates of CRE, 51 isolates were Klebsiella pneumoniae; 10 isolates were Escherichia coli; 15 isolates were other Enterobacterales. The 76 CREs were mainly isolated from urine, sputum and blood samples. The distribution rate of ICU was the highest (55.26%). The 76 CREs showed low resistance rates (0%, 1.33%, 18.42%) to colistin, tigecycline and ceftazidime-avibactam. The resistance rates to amikacin and fosfomycin were <45%, and the resistance rates to other drugs were >97%. The detection rate of KPC-2 carbapenemase was the highest (85.33%). The ST11 CRKP producing KPC-2 carbapenemase accounted for the highest proportion (62.75%), mainly distributed in the ICU (62.50%). Southern blot hybridization showed that blaKPC-2 was mainly located on a plasmid about 90 kb (39/63). Filter mating test showed that blaKPC, blaNDM and blaIMP could be transferred horizontally to recipient bacteria through plasmids. Conclusions:The 76 CRE isolates were only susceptible to a few antibacterial drugs, such as colistin, tigecycline and ceftazidime-avibactam. The production of KPC-2 carbapenemase was the main reason for the resistance of Enterobacterales to carbapenems. KPC-2 carbapenemase-producing ST11 Klebsiella pneumoniae was the main epidemic clone of carbapenem-resistant Klebsiella pneumoniae (CRKP). The 90 kb size plasmid was the main plasmid encoding blaKPC-2 gene. Carbapenemase genes can be transferred horizontally through plasmids. The hospital should strengthen prevention of nosocomial infections to control the clonal prevalence of CRE.

OBJECTIVETo investigate the effects of mouse adipose-derived stem cell conditioned medium (ADSC-CM) on apoptosis of keratinocytes (human epithelial cell line HaCaT) induced by thermal injury in vitro.

METHODS(1) Adipose-derived stem cells (ADSCs) from inguinal adipose tissue of 5 healthy BALB/c mice were isolated, cultured, and purified by collagenase digestion in vitro. The 3rd passage of cells were collected for morphologic observation, detection of expressions of surface markers CD31, CD34, CD45, CD90, and CD105 with flow cytometer, and identification of adipogenic and osteogenic differentiation. (2) HaCaT cells were incubated in water at 51.5 °C for 35 seconds to reproduce thermal injury model, and then the apoptosis rate was detected immediately after injury by flow cytometer. (3) Thermally injured HaCaT cells were divided into routine culture group (RC, cultured with DMEM containing 10% FBS), serum-free group (cultured with serum-free DMEM), 50%ADSC-CM group (cultured with DMEM containing 50%ADSC-CM), and 100%ADSC-CM group (cultured with 100%ADSC-CM) according to the random number table. After 24 hours, apoptosis of HaCaT cells was observed by acridine orange-ethidium bromide (AO-EB) staining; apoptotic rate was determined by flow cytometer; the mRNA and protein levels of Bcl-2 and caspase-3 were respectively determined by real-time fluorescent quantitative RT-PCR technique and Western blotting (protein level was denoted as gray value); the cell cycles were determined by flow cytometer. All above experiments were repeated for 3 times. Data were processed with one-way analysis of variance and LSD- t test.

RESULTS(1) The 3rd passage of cells proliferated well showing fusiform shape similar to fibroblasts. The positive expression rates of CD31, CD34, and CD45 were less than 10.0%, while those of CD90 and CD105 were above 90.0%. The cells could differentiate into adipocytes and osteoblasts. They were identified as ADSCs. (2) Immediately after injury, apoptotic rate of HaCaT cell was (9.8 ± 0.4)%. (3) The number of apoptotic cells was significantly higher in serum-free group than in the other three groups with AO-EB staining. The apoptotic rate of serum-free group [(8.1 ± 1.2)%] was significantly higher than that of 50%ADSC-CM group [(6.0 ± 0.8)%], group RC [(4.6 ± 0.8)%], or 100%ADSC-CM group [(3.1 ± 0.4)%], with t values respectively 3.02, 4.96, 6.60, P values below 0.01. There was no statistically significant difference in apoptotic rate between group RC and 100% ADSC-CM group (t = 1.50, P > 0.05), while statistically significant difference was found between 100% ADSC-CM group and 50%ADSC-CM group (t = 10.21, P < 0.01). (4) The mRNA level of Bcl-2 of serum-free group (0.34 ± 0.08) was significantly lower than that of group RC, 50%ADSC-CM group, and 100%ADSC-CM group (0.98 ± 0.04, 0.77 ± 0.05, 1.06 ± 0.04, with t values respectively 12.87, 8.07, 14.11, P values below 0.01). Compared with that of 100%ADSC-CM group, the mRNA level of Bcl-2 of group RC was slightly decreased (t = 0.08, P > 0.05) and that of 50%ADSC-CM group was significantly decreased (t = 8.08, P < 0.01). (5) The mRNA level of caspase-3 of serum-free group (1.15 ± 0.05) was obviously higher than that of 50%ADSC-CM group (0.72 ± 0.11), group RC (0.41 ± 0.03), or 100%ADSC-CM group (0.38 ± 0.11), with t values respectively 6.93, 13.97, 22.79, P values below 0.01. Compared with 100%ADSC-CM group, the mRNA level of caspase-3 was slightly increased in group RC (t = 0.05, P > 0.05) and significantly increased in 50%ADSC-CM group (t = 4.77, P < 0.01). (6) The protein level of Bcl-2 was significantly lower in serum-free group (0.93 ± 0.04) than in group RC, 50%ADSC-CM group, and 100%ADSC-CM group (1.74 ± 0.06, 1.32 ± 0.05, 1.90 ± 0.04, with t values respectively 20.45, 11.15, 31.38, P values below 0.01). Compared with that of 100%ADSC-CM group, the protein level of Bcl-2 of group RC was slightly decreased (t = 1.33, P > 0.05), but that of 50%ADSC-CM group was obviously decreased (t = 17.30, P < 0.01). (7) The protein level of caspase-3 was obviously higher in serum-free group (0.63 ± 0.08) than in 50%ADSC-CM group, group RC, and 100%ADSC-CM group (0.46 ± 0.03, 0.29 ± 0.08, 0.21 ± 0.03, with t values respectively 3.28, 5.05, 8.46, P values below 0.01). Compared with that of 100%ADSC-CM group, the protein level of caspase-3 of group RC was slightly increased (t = 0.08, P > 0.05), but that of 50%ADSC-CM group was significantly increased (t = 3.52, P < 0.05). (8) Compared with that of serum-free group, the percentage of cells in G2/M phase of each of the other 3 groups was significantly decreased (with t values respectively 6.88, 4.08, 7.28, P < 0.05 or P < 0.01). Compared with that in serum-free group, the percentage of cells in S phase was significantly increased in group RC and 100% ADSC-CM group (with t values respectively 2.67 and 2.40, P values below 0.05). There was no statistically significant difference in the percentage of cells in G0/G1 phase among all groups (F = 0.70, P > 0.05).

CONCLUSIONS100% xenogeneic ADSC-CM can suppress apoptosis of HaCaT cells induced by thermal injury through regulating the expression of Bcl-2 and caspase-3, and accelerate cell cycle progression by ameliorating the retardation of cell growth in G2/M phase, and all these effects may give rise to some potential in the treatment of burn wounds at early stage.

Adipocytes ; Adipose Tissue ; Animals ; Apoptosis ; physiology ; Burns ; Caspase 3 ; metabolism ; Cell Differentiation ; Cell Line ; Cell Proliferation ; Culture Media, Conditioned ; Fibroblasts ; Humans ; In Vitro Techniques ; Keratinocytes ; metabolism ; physiology ; Mice ; Mice, Inbred BALB C ; Real-Time Polymerase Chain Reaction ; Stem Cells