10.3969/j.issn.2095-4344.2013.23.017

The root canals of 95 maxillary second primary molars were explored by 10#file combined with 17%EDTA.The canal orifice was observed and orientated by endoscope.Hand instruments were used for root canal preparation and screw conveyor was used to fill the ca-nal with Vitapex paste.MB2 was found in 14 of the 95 molars(14.7%).MB2 orifice was usually located mesially along MB-P.

Objective:To investigate the pre-pregnancy vitamin D level and pregnancy outcome in patients with unexplained recurrent spontaneous abortion (URSA).Methods:A prospective study was performed in 4 534 patients with URSA from May 2017 to April 2019 at Shanghai First Maternity and Infant Health Hospital Affiliated to Tongji University. The serum Vitamin D levels was obtained before pregnancy. Pregnancy complications and newborns outcomes were recorded after pregnancy.Results:The serum vitamin D level of patients with URSA before pregnancy was (42.22±16.27)nmol/L, and the proportions of vitamin D deficiency, insufficiency and sufficiency were 72.3%, 24.0 %, and 3.7%, respectively. The Vitamin D level was positively correlated with age ( P<0.05); The age of vitamin D<50 nmol/L group was lower than that of vitamin D≥50 nmol/L group ( P<0.05); patients with vitamin D<50 nmol/L had higher proportion of spontaneous abortions ≥3 times than those with the vitamin D≥50 nmol/L ( P<0.05); The level of vitamin D was negatively correlated with the ratio of CD3 + CD4 + T cells in peripheral blood ( P<0.05); In multivariate logistic regression analysis, the final model adjusted for age, abortion frequency and season. The risk of pregnancy failure was increased in vitamin D <50 nmol/L group [30.6%(76/248) vs 17.9%(12/67), χ 2=3.67, P=0.02], OR=2.02(95% CI: 1.02-3.9); In the group of vitamin D<50 nmol/L before pregnancy, the risk of newborns entering NICU was increased, OR=3.16(95% CI: 1.15-8.65). Conclusions:Vitamin D deficiency is prevalent in URSA patients before pregnancy, which correlates with the times of previous spontaneous abortions and recurrent pregnancy failure. Vitamin D deficiency before pregnancy is one of the high-risk factors for URSA.

BACKGROUND:It is necessary to carry out multiple operations to remove the scar in patients with large area of scar, and whether the scar tissue can be recycled has become the focus of the study. OBJECTIVE:To compare the tissue structure, biomechanical properties and biocompatibility of the acel ular dermal matrix of mature scar tissue and normal skin. METHODS:The acel ular dermal matrix was prepared from the human mature scar tissue and the normal skin around the scar. Subsequently, histological and scanning electron microscope observations were performed, and biomechanical properties were detected using universal tensile testing machine. Then, the acel ular dermal matrix from mature scar tissue and normal skin was co-cultured with fibroblasts for 10 days, respectively, and the cel growth curve was drawn. Additional y, the acel ular dermal matrix from mature scar tissue and normal skin was subcutaneously implanted into the dorsal tissue of Sprague-Dawley rats, respectively and histological observation was conducted at 4, 8 and 12 weeks after implantation. RESULTS AND CONCLUSION:There were many gaps but no cel ular components in the acel ular dermal matrix, in both two groups. Col agen fibers of the acel ular dermal matrix derived from mature scar were looser than that of the normal skin, and arranged slightly irregularly;the biomechanical properties of the acel ular dermal matrix derived from mature scar were similar to that of the normal skin, which exhibited appropriate flexibility and strength. There was no significant difference in the growth state of the two kinds of acel ular dermal matrix, and the growth curve was basical y consistent. After 4 weeks of implantation, more inflammatory cel s infiltration could be found in the mature scar group, and in contrast, only a few inflammatory cel s infiltration appeared in the normal skin group, These inflammatory reactions disappeared with time in both two groups. Besides, col agen fibers arranged in neat, and smal vessels grew into the implants in both two groups. In conclusion, the tissue structure, biomechanical properties and biocompatibility of the acel ular dermal matrix derived from scar tissue are almost consistent with those of the human normal skin.

Objective To investigate the efficacy of the clinical pathway introduced in children with bronchiolitis.Methods Based on a retrospective study,the duration of hospital stay,hospital expenses,antibiotics usage rates,curative rate and nosocomial infection rate were compared between 181 bronchiolitis patients (the clinical pathway group) managed according to clinical pathway and other 122 bronchiolitis patients (the control group).The variation of clinical pathway was analyzed in the clinical pathway group as well.Results The duration of hospital stay in clinical pathway group [4.96 (4.00,6.00) days] was significantly shorter than that in the control group [5.81 (4.82,7.00) days],and the difference was significant (Z =3.137,P ＜ 0.05).The hospital expenses [3 701.23 (3 124.50,4 396.19) yuan] in clinical pathway group were significantly lower than that in the control group [3 954.22 (3 325.07,4 679.66) yuan],and the difference was significant (Z =2.042,P ＜ 0.05).The antibiotics usage rate (20.44％)(37/181 cases) in clinical pathway group was significantly lower than that in the control group (40.16％) (49/122 cases),and the difference was significant (,x2 =13.945,P ＜ 0.05),and the curative rate (93.92％) (170/181 cases)in clinical pathway group was significantly higher than that in the control group (86.89 ％) (106/122 cases),and the difference was significant (x2 =4.447,P ＜ 0.05).The nosocomial infection rate (4.42％) (8/181 cases)was also lower than that in the control group (10.66％,13/122 cases),and the difference was significant (x2 =4.393,P ＜0.05).The variation rate of clinical pathway was 48.62％ (88/181 cases) in clinical pathway group.Conclusions The curative rate is improved and the duration of hospital can be shortened if the clinical pathway is introduced in bronchiolitis children,with less the hospital expense.Moreover,the antibiotics usage rates and the nosocomial infection rate are reduced in the implementation process of clinical pathway which is worthy to be put in use.However,there is a high variation rate in the clinical pathway.It should be interposed and administrated appropriately in dinical treatment for clinic.

Objective To analyze the risk factors for hypertension in patients with chronic kidney disease stage 5(CKD5) . Methods The basic information of 390 CKD5 patients complicated with hypertension was collected for univariate analysis ,in‐cluding gender ,age ,primary disease ,dialysis method ,body mass index(BMI) ,complications(hyperlipidemia ,high uric acid ,car‐diac insufficiency) ,level of education ,parathyroid hormone(PTH)level.Univariate variables that showed statistical significance were then subjected to the multivariate analysis(Logistic regression)to identify the risk factors for hypertension in CKD5 pa‐tients.The defined daily dose(DDD)that satisfied the criteria interms of different stages was evaluated.Results Overall hyper‐tension control rate was 22.8%.Univariate analysis showed that the following variables were significantly associated with hy‐pertension in CKD5 patients :>40 years old ,male ,diabetic nephropathy ,hypertensive nephropathy ,hemodialysis ,hyperlipemia , high uric acid level ,and high PTH level(P<0.05).Logistic multivariate analysis showed that diabetic nephropathy ,hyperlipi‐demia ,high PT H level were the independent risk factors for hypertension in patients with CKD5.In hypertension segmented standard ,there was no difference in the DDD between stage 0 and 1(P>0.05) ,and DDD at stage 2 and 3 was increased signifi‐cantly when compared with that at 0 and 1 standard(P<0.05).Conclusion Overall hypertension control rate is very low in pa‐tients with CKD5.Diabetics ,hyperlipidemia ,high PTH level are independent risk factors for hypertension in patients with CKD5.

Background and purpose:EPHA2 has been reported to enhance the proliferation of gastric cancer cells through promoting CyclinD1 expression and cell cycle progression. However, the underlying mechanism that EPHA2 promotes CyclinD1 expression and cell cycle progression is still poorly understood. Akt/mTOR signaling pathway has been reported to enhance the proliferation of gastric cancer cells by promoting CyclinD1 expression and cell cycle progression, and some studies have shown that EPHA2 can activate Akt/mTOR signaling pathway. Based on this, this study investigated whether EPHA2 promoted gastric cancer SGC-7901 cell proliferation through enhancing Akt/mTOR signaling pathway.Methods:Western blot was used to determine the effect of EPHA2 overexpression or knockdown on the phosphorylation of Akt and mTOR in SGC-7901 cells. SGC-7901-NC infected with control lentivi-rus and SGC-7901-EPHA2 cells with EPHA2 overexpression were treated with DMSO, MK2206 (an Akt inhibitor) and RAD001 (a mTOR inhibitor) for different time periods, respectively. Cell proliferation was detected using the CCK8 assay. Cell cycle was detected using lfow cytometry, and the expression of CyclinD1 was determined by Western blot. Results:Overexpression of EPHA2 enhanced Akt/mTOR signaling pathway in SGC-7901 cells, and silencing EPHA2 in SGC-7901 cells inhibited Akt/mTOR signaling pathway. MK2206 and RAD001 antagonized the promoting effect of EPHA2 on the proliferation, S-phase and CyclinD1 expression of SGC-7901 cells, respectively.Conclusion:EPHA2 promotes SGC-7901 cell proliferation through enhancing Akt/mTOR signaling pathway. Akt inhibitor or mTOR inhibi-tor could be an effective treatment strategy for gastric cancer patients overexpressing EPHA2.

Objective To explore the effects of adipose tissue-derived stem cells (ADSCs) on the proliferation and invasion of pancreatic cancer cell lines in vitro.Methods ADSCs were isolated and co-cultured with pancreatic cancer cells.The proliferation and invasion of pancreatic cancer cells was tested by CCK-8 test kit.Stromal cell derived factor (SDF-1),vascular endothelial growth factor(VEGF),matrix metalloproteinase(MMP-9)and transforming growth factor-β (TGF-β1)in the culture medium were assayed by ELISA.The expression of cytokines in pancreatic cancer cells and ADSCs were detected by qRT-PCR.CCK-8 test kit was used to measure the AMD3100 regulation for the co-culture of ADSCs and pancreatic cancer cells.Results ADSCs can promote the proliferation and invasion of pancreatic cancer cells (all P ＜0.05); more SDF-1,VEGF,MMP-9 and less TGF-β1 was secreted by ADSCs than by pancreatic cancer cells(SDF-1:1100±100 vs.0 vs.0,F=389.134,P＜0.01;VEGF:140±4 vs.99±5 vs.93 ±4,F=174.102,P ＜0.05;MMP-9:61.8 ±4.2 vs.43.5 ±2.8 vs.54.5 ±3.0,F=76.279,P＜0.05;TGF-β1:20.6 ±3.0 vs.35.6 ±2.6 vs.41.3 ±5.5,F =79.338,P ＜0.05).ADSCs promote the expression of VEGF and MMP-9 in pancreatic cancer cells(VEGF:63.7 ±5.9 vs.50.6 ±4.1,t =7.536,P＜0.05; MMP-9:mRNA(55.8±3.6 vs.42.7 ±3.1,t =8.279,P＜0.05).AMD3100 significantly downregulates these growth-promoting effects of ADSCs on pancreatic cancer cells(SW1990:1.539 ±0.140 vs.1.361±0.066,t=2.835,P＜0.05;PANC-1:1.376±0.100 vs.1.281±0.031,t=2.860,P＜0.05).Conclusions ADSCs promote the proliferation and invasion of pancreatic cancer cells.

Objective:To study the neuroprotective effect of nobiletin on the symptoms of postoperative cognitive impairment (POCD) induced by sevoflurane inhalation.Methods:Twenty-four aged SD rats (12 female mice and 12 male mice) were divided into three groups randomly: control group ( n=8), surgery group ( n=8) and surgery + nobiletin group ( n=8), with 4 females and 4 males in each group.The rats in surgery group and surgery+ nobiletin group were given normal saline(0.1 ml/10 g, once a day) and nobiletin(100 mg/kg, once a day) intragastrically for 6 weeks.Then the rats were anesthetized by sevoflurane and treated by abdominal exploration surgery, and then continued gavage for 1 week.The rats in control group were given normal saline(0.1 ml/10 g, once a day) intragastrically for 7 weeks without anesthesia or surgery.Sevoflurane inhalation anesthesia and abbreviated laparotomy were not done for control group.Morris water maze and open field experiment were used to measure the memory and cognitive ability and the independent exploration ability respectively.The changes of α-band electroencephalogram (EEG) were recorded by multi-channel physiological signal acquisition and processing system.The concentration of S100β, a marker of neurological impairment was detected by ELISA.Western blot was used to detect the expression level of IBA-1 in microglia.SPSS 20.0 software was used to analyze the data. Results:There were no significant differences in Morris water maze, positioning cruise test and open field test among the groups before operation (all P>0.05). The differences were statistically significant among the groups 7 days after operation (all P<0.05). Compared with the control group (the escape latency, path length and cross platform times were ((20.37±1.11)s, (552.37±14.19)cm, (6.75±0.43)times respectively), the escape latency ((40.87±2.03)s) and path length ((1 258.62±19.53)cm) of rats in surgery group were significantly longer (both P<0.01), and the cross platform times ((2.12±0.33)times) significantly reduced ( P<0.01). The differences between surgery + nobiletin group ((22.37±1.11)s, (584.50±10.90)cm, (6.62±0.48)times) and control group were not significant (all P>0.05). The open field experiment showed that the movement distance, the crossed square number, and activity times in surgery group ((1.78±0.55) m, (4.75±0.50), (14.87±0.33) times) decreased significantly compared with those in control group ((3.73±0.07) m, (11.10±0.78), (51.12±0.78) times, all P<0.01). No significant difference was found between surgery + nobiletin group ((3.76±0.07)m, (10.75±0.66), (50.75±0.43)times) and control group(all P>0.05). Before anesthesia, there was no significant difference in the power ratio of α-band among the three groups ( P>0.05), but the differences during anesthesia and operation were significant ( F=72.58, 101.50, P<0.01). During anesthesia and operation, the power ratio of α-band in anesthesia and in surgery group (2.51±0.04, 2.14±0.03) were significantly lower (both P<0.01) than those in control group (3.49±0.03, 3.49±0.03), while there was no obvious changes (both P>0.05) in the surgery + nobiletin group (3.50±0.04, 3.51±0.04). There were significant differences in Bcl-2 protein expression and caspase 3/7 protein activity among the three groups ( F=5.21, 7.84, P<0.01). Compared with control group (1.00±0.02, 1 557.46±3.63), Bcl-2 of rats in the surgery group(0.40±0.05) were significantly lower and Caspase3/7 expression of surgery group (3 689.58±10.46) was significantly higher (both P<0.01), while the rats in the surgery + nobiletin group had no significant difference in both Bcl-2 level (1.03±0.06) and caspase 3/7 activities (1 805.28±6.17, both P>0.05). The difference of S100 β protein expression was significant among the three groups ( F=490.80, P<0.01). Compared with the control group ((0.18±0.01)μg/L), the concentration of S100β protein in the surgery group ((2.13±0.02)μg/L) decreased ( P<0.01), while there was no significant difference in the surgery + nobiletin group ((0.16±0.01) μg/L, P>0.05). The expression levels of IBA-1 protein ( F=10.83) and TNF-α, IL-1, IL-1β and IL-6 ( F=996.20, 221.40, 73.02, 174.13) were significantly different among the three groups (all P<0.01). The expression level of the neuroglial marker IBA-1 in the surgery group(1.36±0.02) was significantly higher than that in the control group (1.00±0.01, P<0.01), while the surgery + nobiletin group (1.03±0.01) had no significant different compared with control group ( P>0.05). The levels of inflammatory factors, including TNF-α, IL-1, IL-1β and IL-6, in the brain of rats treated with nobiletin ((49.06±3.63)pg/mg, (2.09±0.43)pg/mg, (16.27±0.80)pg/mg, (2.11±0.19)pg/mg) were significantly lower than those in the surgery group((145.10±6.46)pg/mg, (5.67±0.43)pg/mg, (27.88±3.43)pg/mg, (4.74±0.32)pg/mg, all P<0.01). Conclusion:Nobiletin can obviously alleviate POCD symptoms caused by sevoflurane inhalation anesthesia.

Objective To investigate the maturation status of monocytes and their responses to the stimulation of toll like receptor (TLR) ligands during primary HIV-1 infection, and to further understand the correlation between functional status of monocytes and disease progression during primary HIV -1 infection. Methods Peripheral blood mononuclear cells ( PBMCs) were collected from 35 subjects with primary HIV-1 infection and 13 HIV-negative healthy subjects to isolate monocytes .Monocytes were stimulated with LPS and Pam3CSK4, respectively, and cultured for 20 hours.The expression of activaion/inhibitory markers on monocytes were analyzed by flow cytometry before and after stimulation .The secretion of proinflammatory cy-tokines ( IL-1β, TNF-αand IL-6) by stimulated monocytes were detected by ELISA .Results The expres-sion of activation markers CD80, CD86, CD40 and inhibitory marker PD-L1 on monocytes were increased in subjects with primary HIV-1 infection (P<0.001 except for CD86 P=0.01).The level of CD40 was posi-tively correlated with viral load in plasma (P<0.001, R=0.553).Compared with control group, primary HIV-1 infection group showed a less increase in the expression of HLA-DR, CD80, CD86 and PD-L1 on monocytes after stimulation with LPS and Pam3CSK4 (P<0.001), but the secretion of proinflammatory cyto-kines TNF-α(LPS:P=0.004, Pam3CSK4:P=0.012) and IL-6 (LPS:P=0.006) were enhanced in mono-cytes from patients with primary HIV-1 infection.Conclusion Monocytes were activated during primary HIV-1 infection.They secreted higher level of proinflammatory cytokines after stimulation with TLR ligands , indicating monocytes might play a role in microbial translocation and immune activation during HIV -1 infection .