Aim To explore the effect of total flavonoid extract(TFE)of Ampelopsis megalophylla on the pro-liferation and apoptosis of human breast cancer cells and its mechanism in autophagy inhibition.Methods For human cervical cancer cell Hela,human lung cancer cell A549,human liver cancer cell SMMC-7721,human breast cancer cell MCF-7,MDA-MB-231 and human normal liver cell L-02,MTT method was used to select sensitive cell lines.The inhibitory effect of TFE combined with autophagy inhibitor 3-methylade-nine(3-MA)on sensitive cell proliferation was detec-ted using MTT assay.The morphological changes of cells were observed using transmission electron micros-copy and Hoechst 33258 single staining method.The changes in cell apoptosis rate were detected using An-nexin V-FITC/PI dual staining method.The expression levels of apoptosis related proteins and pathway pro-teins(death receptor pathway,mitochondrial pathway,endoplasmic reticulum stress pathway)were detected uisng Western blot.The expression of the key protein Cyt-c in mitochondrial pathway was determiend by im-munofluorescence,and the autophagy agonist rapamy-cin was selected for reverse validation.Results TFE could inhibit the proliferation of human breast cancer cells in a concentration-dependent manner,and MCF-7 cells were sensitive cell lines.Compared with the TFE group,the TFE+3-MA group significantly increased the inhibition rate of MCF-7 cells at 24,48,and 72 h(P＜0.01).The number of cells decreased,the gap increased,the number of apoptotic bodies increased,and the apoptosis rate increased(P＜0.01).The ex-pression levels of Bax/Bcl-2(P＜0.01),cleaved-caspase3(P＜0.01),Cyt-c(P＜0.05),FADD,and cleaved-caspase-12 all increased,and the expres-sion of apoptotic protein Cyt-c in nucleus increased.The fluorescence of the TFE+RA group decreased,re-versing the mitochondrial pathway apoptosis induced by TFE.Conclusions TFE can significantly inhibit the proliferation of human breast cancer cells.When inhib-iting autophagy,it may promote the apoptosis of MCF-7 cells through the mitochondrial pathway,and activa-ting autophagy can reverse apoptosis.

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.
3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics* ; Child ; China/epidemiology* ; Cryptorchidism/genetics* ; Disorders of Sex Development/genetics* ; Female ; Genital Diseases, Male ; Genotype ; Humans ; Hypospadias/genetics* ; Male ; Membrane Proteins/genetics* ; Penis/abnormalities* ; Phenotype ; Retrospective Studies ; Steroid 21-Hydroxylase/genetics*

Objective:To clone cellulose synthase-like(Csl)gene from ethnic medicinal plant Ampelopsis megalophylla,and analyze its sequence by bioinformatics. Method:Specific primers were designed for AmCsl gene sequences obtained from A. megalophylla transcriptome sequencing data. The full-length cDNA of AmCsl gene was amplified by PCR using cDNA of young leaves as template,and TA clone and sequencing was performed. The sequence was analyzed by bioinformatics. Result:The full length cDNA was 1 438 bp,containing a 561 bp open reading frame(ORF),and encoding 186 amino acids,the molecular formula of protein was C₁₀₁₁H₁₅₄₇N₂₃₃O₂₅₇S₁₀,the theoretical relative molecular weight was 22.40 kDa,the theory isoelectric point(PI)was 7.59,and the aliphatic index(AI)was 116.88.There was a transmembrane region and no signal peptide,which may be located in the endoplasmic reticulum,the average hydrophobic coefficient was 0.670,and the instability index was 42.56.It belonged to a hydrophobic unstable protein. The conserved domain contained a cellulose synthase,and the secondary structure mainly was dominated by α-helix. Amino acid sequence alignment and phylogenetic tree analysis showed that AmCsl had a high homology with Vitis vinifera. Conclusion:The full length of AmCsl gene was obtained for preliminary bioinformatics analysis,which laid a necessary foundation for further study on the accumulation of polysaccharides and the regulation of biosynthesis.

OBJECTIVE Cloves(Syzygium aromaticum L.)have been used as both a spice and a traditional Chinese medicinal herb for thousands of years. However, relatively little is known about its potential anticancer activity and mechanisms.In this study,we investigated the in vitro and in vivo anti-tumor effects and mechanisms of water extract of cloves(WEC)against colorectal cancer. METHODS MTS assay and Colony-formation assay were used to detect the anti-tumor activity of WEC on HT-29 cells.The in vivo anti-tumor effect of WEC was detected in a subcutaneous transplantation tumor model of human HT-29 cells.Autophagy was detected by flow cytometry and the expressions of autophagy related proteins(Beclin-1 and LC-3a/b)were determined by western blot. RESULTS MTS result showed that WEC significantly inhibited the viability of HT-29 cells,with the IC50values of 150 μg·mL-1.The colony-formation assay showed that the WEC significantly suppressed colon cancer cells proliferation.WEC also exhibited significant antitumor activity in tumor bearing nude mice. Flow cytometry result showed that WEC significantly induced autophagy, and the averaged relative values of fluorescence intensity were 206,251,341 and 356 in cells treated with 0,100,150 and 200 μg·mL-1WEC for 48 h.Western blot result showed that WEC treatment significantly increased Beclin-1 expression and ratios of LC3-II/LC3-I. CONCLUSION These result showed that WEC inhibited the growth of colon tumor both in vitro and in vivo, which might be related with autophagy induction, and WEC has potential to be developed as a novel anticancer agent for the treatment of colon cancer.

OBJECTIVE Zn-doped CuO nanocomposites (Zn-CuO NPs) are novel nanoparticles synthesized by our research group.In this study,we assessed the in vitro and in vivo antitumor effects of Zn-CuO NPS on pancreatic cancer cells,as well as the potential mechanisms. METHODS MTS assay was used to detect the effects of Zn-CuO NPS on proliferation pancreatic cancer cells(Panc-mia and Aspc-1). The in vivo antitumor effects of Zn-CuO NPs were detected by xenografts model in nude mice. The effects of Zn-CuO NPS on autophagy were detected bytransmission electron microscopy (TEM) andflow cytometry. Autophagy related proteins were detected by Western blotting. RESULTS Zn-CuO NPS significantly inhibited the proliferation of Panc-mia cells and Aspc-1 cells.In vivo experi-ments showed that Zn-CuO NPS significantly inhibited the tumor growth in nude mice without affecting the body weight of the mice. TEM and flow cytometry showed that Zn-CuO NPS induced autophagy, and significantly increased the number of autophagosome.Western Blot showed that Zn-CuO NPS alterd the expression of autophagy related proteins,such as AMPK,mTORand Beclin-1.Also,AMPK inhibitor could significantly reduce Zn-CuO NPS-induced autophagy pathwayas analyzed byWestern blotting. CONCLUSION The findings suggested that Zn-doped CuO nanocomposites inhibited the in vitro and in vivo growth of pancreatic cancer by inducing autophagy through AMPK/mTOR pathway.

OBJECTIVE Granulin A (GRN A), a cytokinesis protein, is derived from proteolysis of progranulin. The previous study in our laboratory has shown that GRN A is able to inhibit cancer cell growth significantly. This study aimed to investigate the effect of combination of GRN A and cisplatin on in vitro and in vivo on the growth of hepatocellular carcinoma. METHODS The in vitro and in vivo antitumor effects of combination of GRN A and Cisplatin were evaluated with MTS assay and subcuta-neous transplantation tumor model.Chou-Talalay method was used to calculate the combination index (CI). Colony formation assay and flow cytometry were used to detect the effects of GRN A on apoptosis. The expression of apoptosis-related proteins were detected by Western blot. RESULTS MTS assay showed that GRN A significantly inhibit hepatocellular carcinoma cells growth with the IC50of 5.6 μmol·L-1, and GRN A combined with cisplatin synergistically inhibit hepatocellular carcinoma proliferation, with the CI<1.The colony-formation assay showed that GRN A significantly enhanced the inhibitory effects of cisplatin on cellular anchorage-independent growth. Flow cytometry showed that GRN A combined with cisplatin synergistically induced apoptosis,with the apoptotic rates of 5.87%,32.74%,35.67% and 67.15% in control, GRN A, Cisplatin, and combination of GRN A and Cisplatin groups, respectively. Western blot confirmed that the two drugs synergistically changed the expressions of proteins related to apoptosis.In vivo experiment indicated that combination of GRN A and cisplatin significantly suppressed tumor growth compared with single drug treatment groups.CONCLUSION The combination of GRN A and cisplatin resulted in synergistic antitumor effects against hepatocellular carcinoma both in vitro and in vivo.

Objective To explore the sedation effect of dexmedetomi-dine and bispectral index for the treatment of intensive care unit( ICU) patients with mechanical ventilation.Methods A total of 58 postopera-tion ICU patients needing mechanical ventilation were randomized into treatment group and control group. Treatment group was intravenous injected dexmedetomidine 0.5 μg? kg-1 , and 0.2 μg? kg -1? h-1 in-travenous infusion.Control group was intravenous infusion loading dose of midazolam 0.06 mg? kg -1 , intravenous infusion 0.4 mg? kg-1? h-1 to maintain sedation.The arterial pressure ( MAP) , respiratory rate ( RR) , pulmonary cardiac blood oxygen saturation ( SpO2 ) , heart rate ( HR ) , Ramsay score, bispectral index ( BIS) , adverse reactions and complica-tions at different times were monitored.Results MAP, RR, HR and SpO2 at each time were no difference after sedation ( P>0.05).RR and HR decreased obviously compared with before sedation ( P<0.05). Compared with before sedation, BIS value significantly decreased, Ram-say score was significantly higher ( P<0.05 ) .After sedation, the BIS value and Ramsay score had no difference between the two groups (P>0.05).Delirium,beckoning too moderate and hypotension incidence in treatment group were lower those of con-trol group ( P<0.05 ) , duration from ICU arrival to off time and diastasis were significantly shorter than that of control group( P <0.05 ) .Conclusion Dexmedetomidine has good sedation effect for patients with mechanical ventilation with reduced adverse reactions and complications.

Objective To study the optimal waist circumference (WC) cut-off values for identifying metabolic risk factors in middle-aged and elderly subjects in Shandong Province of China.
Methods A total of 2 873 men and 5 559 women were included in this cross-sectional study. Metabolic syndrome (MetS) was diagnosed according to the definition of Chinese Diabetes Society in 2004. The relation between WC and MetS was analyzed by multivariate logistic regression analysis. The optimal WC cut-off values were identified using the area under the ROC curve and the different diagnostic criteria for central obesity were compared.
Results The WC was the risk factor for MetS independent of BMI, blood glucose, blood lipid, and blood pressure. The optimal WC cut-off value was 83.8 cm and 91.1 cm for identifying MetS in women and men, respectively. Compared with 80 cm and 85 cm for women and men, 85 cm and 90 cm had a higher Youden index for identifying all metabolic risk factors and MetS in women and men.
Conclusion The appropriate WC cut-off value is 85 cm and 90 cm for identifying central obesity and MetS in women and men in Shandong Province of China.

OBJECTIVETo evaluate clinical therapeutic effect of post-stroke shoulder pain treated by acupuncture combined with Tuina.

METHODSThree hundred cases of post-stroke shoulder pain were randomly divided into an acupuncture and Tuina group and a rehabilitation group by double-center randomized controlled clinical trial method. In acupuncture and Tuina group, normalized electroacupuncture and Tuina therapy were applied, that was electroacupuncture at main points, such as Chize (LU 5), Quze (PC 3), Shaohai (HT 3), Jianyu (LI 15), Jianliao (TE 14) and Jianjing (GB 21),etc., combined with traditional Tuina manipulations; in rehabilitation group, the rehabilitation methods such as the electrostimulation through nervus cutaneus and the squeezing and stabilizing manipulations of Proprioceptive Neuromuscular Facilitation (PNF), etc. were applied. The treatment courses of both groups were 6 weeks. The main therapeutic effect indices were the Assessment Face Scale (AFS) for pain when shoulder was in passive motion and the Fugl-Meyer Motor Assessment for upper limbs active function; the secondary indices were the moditied Rankin Scale (mRS) and the clinical incidences of shoulder-hand syndrome of hemiplegia and shoulder joint subluxation of hemiplegia.

RESULTSAfter 6 weeks treatment and 12 weeks follow-up, AFS score, Fugl-Meyer motor assessment of upper limbs active function and mRS evaluation in acupuncture and Tuina group were more obviously improved than those in rehabilitation group (P < 0.05, P < 0.01). Although the clinical incidences of shoulder-hand syndrome of hemiplegia and shoulder joint subluxation of hemiplegia in acupuncture and Tuina group was equal to those in rehabilitation group [3.55% (5/141) vs 8.45% (12/142), 1.42% (2/141) vs 5.63% (8/142), both P > 0.05], the data indicated that there was a superiority tendency in acupuncture and Tuina group.

CONCLUSIONThe combined therapy of electroacupuncture and Tuina is a normative manipulation, and the therapeutic effect is satisfying for post-stroke shoulder pain, superior to that of comprehensive rehabilitation treatment.

Aged ; Amobarbital ; Combined Modality Therapy ; Drug Combinations ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Range of Motion, Articular ; Secobarbital ; Shoulder Pain ; etiology ; physiopathology ; therapy ; Stroke ; complications

Objective To investigate the clinicopathologic relationship between serum platelet derived growth factor-BB(PDGF-BB)and IgA nephropathy in children.Methods The level of serum PDGF-BB was detected by double antibody enzyme linked immunosorbent assay(ELISA) in 15 cases healthy children and 30 cases IgA nephropathy.According to patholgical degree constituted by WHO in 1982,the IgA nephropathy group divided into 5 degrees:Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ(Ⅰ,Ⅱ were light patholgic change group;Ⅲ,Ⅳ were moderate patholgic change group;Ⅴ was severe patholgic change group).The serum PDGF-BB in IgA nephropathy group and none-IgA nephropathy group,and in different renal pathology type IgA nephropathy group were analyzed.Data were analyzed by using SAS 6.12 software.Results The level of serum PDGF-BB were(247.35?55.79) ng/L in control group and(869.16?200.73) ng/L in IgA nephropathy group.It was higher in IgA nephropathy group than in control group,the difference was significant(P