BACKGROUND:Human periodontal ligament stem cels are a kind of mesenchymal stem cels that have self-renewal and multidifferentiation potential. Previous studies have showed that human periodontal ligament stem cels can differentiate into osteoblast-like cels or adipocyte-like cels under appropriate induction. Yet few studies have focused on the expression level of parathyroid hormone 1 receptor which wil affect the osteogenic potential of Human periodontal ligament stem cels. OBJECTIVE:To examine the expression level of parathyroid hormone 1 receptor between human periodontal ligament stem cels and human periodontal ligament cels and to discuss the role of parathyroid hormone 1 receptor in osteogenic differentiation. METHODS:By using magnetic-bead cel sorting, we separated and identified the human periodontal ligament stem cels and human periodontal ligament cels. We examined and compared the mRNA expression level of parathyroid hormone 1 receptor in human periodontal ligament stem cels and human periodontal ligament cels by Real-Time PCR. Osteoblastic differentiation was examined throughin vitro matrix mineralization by alizarin red staining and alkaline phosphatase assay. RESULTS AND CONCLUSION: Positive immunomagetic sorted cels were positive for STRO-1, CD146, Vimentin, indicating that they were periodontal ligament stem cels. Parathyroid hormone 1 receptor was expressed in human periodontal ligament stem cels and mainly located in cel membrane and cytoplasm which were similar to human periodontal ligament cels and MG63 cels. The expression of parathyroid hormone 1 receptor in human periodontal ligament stem cels was 3.7 times higher than that in human periodontal ligament cels, which was similar to that in MG63 cels. After osteogenic induction, human periodontal ligament stem cels showed a higher expression of parathyroid hormone 1 receptor and osteoblast-related genes as wel as the activity of osteoblast alkaline phosphatase and mineralization compared to human periodontal ligament cels. Our data showed that parathyroid hormone 1 receptor was higher in human periodontal ligament stem cels than human periodontal ligament cels and the expression was related with osteogenic differentiation, suggesting that human periodontal ligament stem cels display a higher potency of osteogenic differentiation and act as seed cels with a vast application prospect in oral tissue engineering.

AIM: To investigate the variations of heme oxygenase-1/carbon monoxide system in lung ischemia-reperfusion injury and effects of puerarin on the system.METHODS: The unilateral lung ischemia-reperfusion model was replicated in vivo.Rabbits were randomly divided into three groups(n=10 in each): control group,ischemia-reperfusion(I-R) group and puerarin group.The blood specimens collected at times before ischemia,ischemia 1 h,reperfusion 1 h,3 h and 5 h were tested for the contents of carboxyhemoglobin(COHb) and cyclic guanosine monophosphate(cGMP).The lung tissues sampled at 5 h after reperfusion were assayed for wet/dry weight ratio(W/T),the injured alveoli rate(IAR) and the activity of HO-1.The changes of ultrastructure were observed under electron microscope.The tissue slides were also stained by immunohistochemistry(IHC) and in situ hybridization(ISH) to detect the expression of HO-1.RESULTS: The plasma content of COHb and cGMP in I-R and puerarin group increased in a time-dependent manner after I-R compared with control group,but the increment in puerarin group was higher than that in I-R group(P

AIM:To explore the relationship between apoptosis in the lung tissues and lung ischemia/reperfusion injury,and to observe the effects of human thioredoxin (hTrx) on apoptosis in lung ischemia/reperfusion injury. METHODS:The single lung in situ ischemia/reperfusion animal model was used. Eighty four Wistar rats were randomly divided into control group (control),groups of ischemia for 1 h and reperfusion for different times (IR1h,IR3h,IR5h),and groups of IR + human thioredoxin treatment (IR1h + hTrx,IR3h + hTrx and IR5h + hTrx). Transmission electron microscope (TEM),terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and immunocytochemistry techniques were used to observe apoptosis,apoptosis signal-regulating kinase 1 (ASK1) and expression of Bcl-2 and Bax in various phases of lung ischemia/reperfusion. RESULTS:Cell apoptosis in lung tissues was significantly high,ASK1,Bcl -2 and Bax protein were upregulated in lung tissues of lung ischemia/reperfusion injury as compared to control (all P

Objective To investigate the structure of deoxyhypusine synthase(DHS)in Saccharomyces cerevisiae(Dys1)and unravel the molecular mechanism of hypusine lysine modification,providing a theoretical basis for the treatment of highly proliferative diseases such as human immunodeficiency virus type 1(HIV-1)replication.Meth-ods Using the E.coli BL21 expression system,an in vitro expression vector was constructed and used to express the protein of Dys1.Dys1 protein samples were purified using methods such as affinity chromatography and molecu-lar sieving to achieve protein purification and isolation.The crystals of Dys1 were obtained using the crystallized so-lution containing 6％Polyethylene Glycol(PEG)8000,0.1 mol/L N-2-hydroxyethylpiperazine-N-ethane-sulphoni-cacid(Hepes)pH 6.5,and 8％ethylene glycol.The crystal structure of Dys1 was resolved at a resolution of 2.8 ? using X-ray crystallography.The structural analysis was performed with CCP4i and Coot software.Results The overall structure of Dys1 was a tetramer,each monomer containing a catalytic site and a cofactor NAD+binding site.The core region of the monomer adopted a Rossmann fold.The amino acid residues involved in the substrate binding sites were highly conserved among eukaryotes.Conclusion The crystal structure of Dys1 is being resolved for the first time.It reveals the binding mode of the cofactor NAD+to the enzyme and confirms that the enzyme functions as a tetramer,with the N-terminus serving as an essential modulator for its catalytic activity.

AIM:To investigate the effect of L-arginine(L-Arg) on expression of bcl-2,bax mRNA during pulmonary ischemia and reperfusion injury(PIRI) in rabbits.METHODS: Single lung ischemia and reperfusion animal model was used in vivo.The rabbits were randomly divided into three groups: sham operated group(sham,n=12),ischemia-reperfusion group(I/R,n=12) and I/R+ L-arginine group(L-Arg,n=12).Changes of several parameters,which included apoptotic index(AI),wet to dry ratio of lung tissue weight(W/D) and index of quantitative assessment of histologic lung injury(IQA),were measured at 300 min after reperfusion in lung tissue.Meanwhile the location and expression of bcl-2,bax mRNA as well as the ratio of bcl-2 mRNA/bax mRNA were observed.The lung tissue was prepared for light microscopic and electron microscopic observation at 60,180 and 300 min after reperfusion.RESULTS: As compared with I/R group,in intima and extima of small pulmonary artery,alveoli,and bronchiole epithelia,the expression of bcl-2 mRNA and the ratio of bcl-2 mRNA/bax mRNA were increased,and the expression of bax mRNA was decreased in L-Arg treatment group.The values of AI,W/D and IQA showed significantly lower than that in I/R group at 180 minutes after reperfusion in lung tissue(P

Objective To explore the molecular genetic characteristics of primary and recurrent glioblastomas (GBMs) from the same patient in vivo, primary glioma stem cells cultured in vitro, and patient-derived xenograft (PDX). Methods (1) The primary and recurrent GBM specimens from one patient during surgical resection were collected; and the expressions of glial fibrillary acidic protein (GFAP), nestin and Ki-67 were detected by immunohistochemical staining; the methylation of O6-methylguanine DNA methyltransferase (MGMT) gene, mutation of isocitrate dehydrogenase (IDH) gene and amplification of epidermal growth factor receptor (EGFR) gene were analyzed. (2) The primary and recurrent GBM stem cells were cultured in vitro and named as SU5-1 and SU5-2 cells, respectively; the expressions of nestin and CD133 were detected by immunohistochemical staining; GFAP expression was detected by immunohistochemical staining after induced differentiation, and the growth curve was detected by CCK-8 assay; Transwell invasion assay was used to detect the invasion ability; cell resistance to temozolomide (TMZ), carboplatin (CBP), cisplatin (DDP) and adriamycin (ADM) was detected by CCK-8 assay; the protein expression of programmed death receptor-ligand 1 (PD-L1) was detected by Western blotting. The rate of PD-L1 positive cells was detected by flow cytometry; genetic testing analysis was as above. (3) The primary and recurrent in situ PDX models in nude mice were established, and the expressions of nestin, GFAP and Ki-67 were detected by immunohistochemical staining. Results (1) As compared with the primary GBM, the recurrent GBM had significantly higher percentages of Ki-67 and nestin positive cells, while statistically lower percentage of GFAP positive cells (P<0.05); genetic analysis showed that there was no mutation in IDH gene in the primary GBM tissues and recurrent GBM tissues; the MGMT gene in the primary GBM tissues was methylated and EGFR gene was not amplified, while the MGMT gene in recurrent GBM tissues was demethylated and EGFR gene amplification was positive. (2) Both SU5-1 and SU5-2 cells expressed nestin and CD133, and GFAP was expressed after induced differentiation; the growth curve showed that the proliferation of SU5-2 cells started earlier than that of SU5-1 cells, the two were equal on the 3rd, 4th, and 5th d, and the proliferation of SU5-1 cells was faster than that of SU5-2 cells from the 6th d; the invasion ability of SU5-2 cells was statistically stronger than that of SU5-1 cells (P<0.05); the inhibition rates of SU5-2 cells treated with 5, 10, and 15 mmol/L CBP, 0.3125, 1.25, and 5 mmol/L DDP, 0.5 and 2 mmol/L ADM, and 125 and 500 mmol/L TMZ were significantly lower than those of SU5-1 cells treated with the same concentrations and same drugs (P<0.05); the protein expression of PD-L1 in SU5-2 cells was higher than that in SU5-1 cells; the positive rate of PD-L1 in SU5-2 cells was statistically higher than that in SU5-1 cells (P<0.05); the results of genetic analysis were consistent with those of the primary and recurrent GBM samples. (3) As compared with those in the primary PDX model, the nestin and Ki-67 expressions were significantly higher and GFAP expression was significantly lower in the recurrent PDX model (P<0.05); the results of genetic analysis were consistent with those of the primary and recurrent GBM samples. Conclusions Genetic differences are detected between primary and recurrent GBMs; recurrent GBM has stronger invasive capacity and multi-drug resistance. The primary stem cells derived from surgical specimens and corresponding PDX models could replicate the molecular genetic characteristics of original tumors, which provide a reliable experimental platform for both tumor translation researches and screening of molecular therapeutic targets.

Objective:To evaluate the short-term influence of bilateral subthalamic deep brain stimulation (STN-DBS) in anxiety symptoms and quality of life in patients with Parkinson's disease (PD).Methods:Thirty-nine PD patients underwent bilateral STN-DBS in our hospital from August 2017 to August 2018 were chosen in our study. Beck Anxiety Inventory (BAI) and Beck Depression Inventory (BDI) scales were performed in these patients before and one month after surgery, and at the last follow-up, respectively. Parkinson's Disease Questionnaire-8 (PDQ-8) was performed before surgery and at the last follow-up. Statistical methods were used to analyze the differences of the above scores at different time points, and correlations between each two improvement degrees of above scores. The participants were subsequently divided into four groups based on preoperative BAI scores: no anxiety group ( n=18), mild anxiety group ( n=10), moderate anxiety group ( n=8), and severe anxiety group ( n=3); the above scales were performed. Results:(1) The BAI scores of 39 patients one month after surgery and at the last follow-up (14 [8, 20] and 9 [3, 14]) were significantly lower than those before surgery (16 [9, 27]), and the BDI scores (8[6, 16]) and PDQ-8 scores (3 [2, 6]) at the last follow-up were significantly lower than those before surgery (15 [8, 21] and 9 [6, 13], P<0.05). (2) Correlation analysis revealed that the improvement degree of BAI scores was positively correlated with that of BDI scores ( r s=0.722, P=0.000), and negatively correlated with preoperative baseline scores of BDI and PDQ-8, respectively ( r s=-0.714, P=0.000; r s=-0.378, P=0.018). (3) The BAI scores in the mild and moderate anxiety groups at the last follow-up were significantly lower as compared with the preoperative baseline BAI scores ( P<0.05). The improvement degree of BAI scores in mild, moderate, and severe anxiety groups was significantly higher than that in the no anxiety group ( P<0.05). Conclusion:The bilateral STN-DBS has considerable benefit effect on anxiety symptoms and health-related quality of life in PD patients during short-term follow-up, suggesting an involvement of STN in the pathogenesis of anxiety in PD.

Objective To evaluate the correlation of CT enhancement parameters with Fuhrman grade in T1 (≤7 cm) clear cell renal carcinoma(ccRCC).Methods From September 2011 to November 2017,102 post-operation cases in our hospital proven to be T1 ccRCC were retrospectively analyzed.There were 71 males and 31 females,with a mean age of (59.1 ± 12.7)years (26 ～79 years),mean body mass index(BMI) of (24.0 ± 2.8)kg/m2 (14.3 ～ 31.6 kg/m2).Tumors of 55 patients were in left kidneys,47 in right kidneys.Fuhrman grade 1 and 2 were defined as low-grade group,meanwhile high-grade group included grade 3 and 4.There were 46 males and 21 females in low-grade group,with a mean age of (59.0 ± 13.2) years,mean BMI of (24.0 ± 2.9) kg/m2.In high-grade group,there were 25 males and 10 females,with a mean age of (58.8 ± 11.8) years,mean BMI of (24.2 ± 2.7) kg/m2.The maximum diameter and tumor enhancement value (TEVX),relative enhancement value (REVX) were measured and calculated.In arterial phase,X =1;in venous phase X =2.The total consumption amount of iodine was recorded.Comparisons of maximum diameter,TEV1,TEV2,REV1,REV2 and the total consumption of iodine between the two different groups were performed.The ROC curves of TEV1,TEV2,REV1,and REV2 were drawn to predict the grade of tumors..Results The TEV1 [(146.1 ± 29.1) HU vs.(100.2 ± 32.1) HU],TEV2 [(98.2 ± 22.9) HU vs.(75.6 ± 25.7) HU],REV1 (1.12 ± 0.24 vs.0.70 ± 0.16),REV2(0.67 ± 0.17 vs.0.54 ± 0.18) between low-grade group and high-grade group had statistical difference (P ＜ 0.05).There was no significant difference in the maximum diameter[(41.8 ± 15.4)mm vs.(45.3 ± 17.0)mm] and the total consumption of iodine [(33.3 ± 5.0)g vs.(34.2 ± 4.4)g] between the two groups (P ＞ 0.05).The area under ROC curve of TEV1,TEV2,REV1 and REV2 were 0.856,0.755,0.901 and 0.728,respectively.REV1 had the highest distinguish efficiency and the best critical value was 0.93.Conclusions The enhancement parameters of T1 ccRCC could contribute to predicting the Fuhrman grade.When the REV1 ≤0.93,the tumor tended to be high-grade tumor(Fuhrman grade 3-4).

OBJECTIVE： To provide reference for perfecting the quality standard revision of TCM preparations in medical institutions， and improving the quality standard of TCM preparations in medical institutions. METHODS： By sorting out the quality standards of TCM preparations in Guangdong provincial medical institution preparation specification， this paper summarized the main problems and put forward suggestions for improvement. RESULTS & CONCLUSIONS： There are 897 kinds of TCM preparations included in the Guangdong provincial medical institution preparation specification. The recent quality standards （2017 edition） have been greatly improved compared with those of the first edition （1985 edition）； however， there are still some problems in the overall quality control of preparations， quantitative control of indicative components， project specificity， source control of medicinal materials， control of medicinal materials not included in the Chinese Pharmacopoeia and quality standards of the same series of varieties. It is suggested that on the premise of considering both advancement and applicability， the quality control of the whole， local and toxic preparations of TCM hospitals should be strengthened， the detection methods with good specificity and reproducibility should be properly updated， the control of key parameters of production process should be strengthened， the quality standard of TCM preparations in medical institutions should be fully improved， so as to provide the products with safe， effective and controllable in quality products in medical institutions.

Objective:To investigate the clinicopathologic features and molecular genetics characteristics of sinonasal tract mucosal malignant melanomas(STMMMs)in elderly patients.Methods:The clinicopathological features, immunohistochemical features and BRAF, C-KIT, NRAS mutations of STMMM in ten elderly patients were retrospectively analyzed.Results:Among the 10 patients, 5 were female and 5 were male.The patients were aged 65-81 years, with an average age of(72.5 ± 8.5)years.The lesions in 7 cases were located in the nasal cavity and paranasal sinuses, and in the other 3 cases were located in the nasopharynx.The morphologies of tumor cells under microscope was complex and diverse, showing plasma cell-like, rhabdomyoblast-like, small cell-like, epithelial-like, and spindle cell-like morphologies.Immunohistochemically, HMB-45 and S-100 were generally positive in 10 cases, and the positive rate of Melan A was 70.0%.The genes detection data showed no mutations in BRAF or NRAS genes in all the 10 cases, while C-KIT exon 11 c. 1666_1667insA mutation was found in one case, and the remaining 9 cases were wild-type for C-KIT.All the 10 cases were followed up for 4~50 months.Three cases survived so far.Conclusions:STMMM in elderly patients are rare and easy to be misdiagnosed.Immunohistochemistry and genetic testing provide guidance for accurate diagnosis and targeted therapy.