Objective:To establish the methods for the determination of Mg,Al,K,Ca,Cr,Cu,Zn,As,Se,Ag,Cd,Sn,Ba,Pb,Na and Hg in water for injection. Methods:The sixteen metallic elements in water for injection were determined by ICP-MS. The collision cell technology ( CCT) was used to minimize the multiple atomic interference, and the matrix effect and signal drift were compensated by using Li, Sc, Ge, In, Ir and Bi as the internal standards and the samples were directly acidified to detect the metallic elements. Results:The detection limit range of the 16 metallic elements were from 0. 009 to 0. 165 ng·ml-1. The calibration curves showed good linearity (r≥0. 999 0). The recoveries were within the range of 80%-120%(n=6). Conclusion:The method is simple, rapid and accurate. It can be used to determine the contents of metallic elements to reduce the potential exceeding limit risk of toxic metal el-ements in water for injection. The methods also can provide reference for the more strict quality evaluation of water for injection.

Objective To study the expression of TFIIB-related factor 2 (BRF2) in hepatocellular carcinoma(HCC) and to determine its clinical significance.Methods 80 HCC patients who received ‘curative' hepatectomy at the Qilu Hospital were studied.Immunohistochemistry analysis was performed to examine the expression of BRF2 and CD34 (microvessel density) in tumor tissues,matched paraneoplastic tissues,and normal liver tissues.Statistical methods were performed to analyse the relationships of expressions of BRF2 and CD34 with clinicopathologic factors.Results BRF2 protein was expressed at a high rate of 61.3％ in HCC tumor,which was significantly higher than that in normal liver tissues and matched paraneoplastic tissues (31.6％,28.8 ％,respectively,P ＜ 0.05).BRF2 was significantly associated with tumor differentiation,number of nodules,tumor encapsulation,TNM stage,and microvessel density.Prognosis of the high expression BRF2 group was poor.The recurrence and metastasis rates in the high expression group were significantly higher than the low expression group (P ＜ 0.05).The survival rate in the high expression group was significantly lower than the low expression group (P ＜ 0.05).Conclusions The expression of BRF2 increased in HCC tissues,and its expression was closely associated with pathological features and prognosis of patients,indicating that it can be used as a predictor in assessing prognosis of patients with HCC.

Root canal biofilm is frequently detected in the canal wall of infected root canal and the root canal with failed root canal therapy. Due to its special structure and diverse composition, root canal biofilm has the ability of the drug tolerance and antiimmunity, which lead to apical periodontitis. This review summarizes the features of the root canal biofilm and latest clinical methods to remove it.

Objective To investigate the expression of PHF8(PHD-finger protein 8) in human hepatocellular carcinoma (HCC) and its clinicopathologic significance.Methods The expression of PHF8 in 60 hepatocellular carcinoma samples as well as their natched paraneoplastic tissues,and 15 normal liver tissues were evaluated by immunohistochemistry.Statistical methods were used to analyse the relationship between the expression of PHF8 and the clinicopathological features of these patients.Results The PHF8-positive expression rate in the HCC samples was 55.0％ and it was significantly higher than that in the paraneoplastic tissues and the normal liver tissucs (16.7％,6.7％,respectively,P＜0.05).The expression of PHF8 was closely related to tumor size,tumor nodular numbers,pathological differentiation and TNM-staging (P＜0.05 for all).The 5-year disease-free survival and overall survival in the PHF8-positive group was significantly lower than that in the negative group (P＜0.05).Conclusions PHF8 was overexpressed in HCC samples,and its expression was closely associated with HCC clinicopathological features and prognosis of the patients.

Objective: : To explore the effect of PD-1 gene knockout by CRISPR/Cas9 system on the proliferation and IFN-γ secretion in human T cells. Methods: : The sequence of sgRNA targeting PD-1 was designed. The PD-1-sgRNA and Cas9 mRNA were synthesized by T7 RNApolymerase in vitro, and then the mixture of PD-1-sgRNAand Cas9 mRNAwas delivered into activated T cells by nucleofection. The efficiency of gene knockout was confirmed by sequencing. The phenotypes of T lymphocytes and the expression of PD-1 after gene knockout were analyzed by Flow cytometry. The proliferation of T lymphocytes was calculated by trypan blue counting. The level of IFN-γ secreted by T lymphocytes was detected by ELISA. Results： ：PD-1-sgRNA and Cas9 mRNA were successfully synthesized in vitro and delivered into T cells by nucleofection. Sequencing technology confirmed that the PD-1 gene sequence was edited and the editing efficiency was 58.3%. The expression of PD-1 on T lymphocyte surface was down-regulated successfully by CRISPR/Cas9 system [(9.6±1.85)% vs (16.2±2.05)%, P<0.05]. The knockout of PD-1 gene did not affect the proliferation and phenotype of T lymphocytes(P<0.05); However, compared with the control group, the level of IFN-γ secreted by T lymphocytes in the PD-1sgRNA group was significantly increased (P<0.01). Conclusion: : CRISPR/Cas9 system can successfully ablate PD-1 gene in human T lymphocytes, which could block the negative regulation of PD-1／PD-L1 and further promote the IFN-γ secretion in T cells.

Objective To compare the clinical efficacy of minimally invasive removal of intracranial hematomas and medical con-servative treatment for hypertensive cerebral hemorrhage .Methods A total of 75 patients with hypertensive cerebral hemorrhage were collected and randomly divided into two groups according to the treatments :38 patients treated with conservative medicine and 37 patients treated with minimally invasive removal of intracranial hematomas based on standard drug therapy .Neurological impair-ment and recovery of self-care ability of patients in both groups before and after three weeks and six weeks of treatment were scored by National Institute of Health Stroke Scale (NIHSS) .Changes of diffusion tensor imaging before treatment and after two weeks of treatment were evaluated by fractional anisotropy value (FA) .Results There was no significant difference between neurological im-pairments of the patients in two groups before treatment .But the recovery degree of functional impairment and muscle nerve in min-imally traumatic treatment group was more significant than those in the conservative medical treatment group after treatments . There was significant difference between two groups (P< 0 .05) .Conclusion Minimally traumatic of hematomas is an effective method of treating hypertensive intracerebral hemorrhage .

BACKGROUND: Based on a mouse model of tibial implantation, some scholars have found that the CaP-coated implant with recombinant human parathyroid hormone (1-34) (PTH(1-34)) shows strong osteogenesis effect at early stage, but this coating has not been applied in the oral environment.OBJECTIVE: To explore the role of local application of PTH(1-34) on immediate implant osseointegration . METHODS: Nine New Zealand white rabbits were randomly divided into two groups (six in experimental group and three in control group). All of the tooth sockets were filled with heterogeneous freeze-dried bone firstly after four incisors of each rabbit were extracted. In the experimental group, a titanium screw with PTH(1-34) loaded CaP coating was implanted into each tooth socket, while in the control group, a titanium screw with only CaP coating was implanted. The animals were executed respectively at 4, 8, 12 weeks after operation, and the intact maxillary and mandibular specimens were harvested and tested by gross observation, bone density analysis, torque test, histologic al observation, X-ray observation.RESULTS AND CONCLUSION: The gray value and maximum torque value of regenerated osseous tissue at different time points in the experimental group were significantly higher than those in the control group (P < 0.05). Within 4-12 weeks after implantation, regenerated and mature bone tissue appeared earlier in the experimental group than the control group. A large amount of new blood vessels were seen in the experimental group at 8 weeks after implantation, while in the control group, there were only few new blood vessels. To conclude, the local application of PTH(1-34) can promote bone formation, improve the implant-bone bonding strength, and enhance the stability of the implant.

BACKGROUND:Nanosilver has significant antibacterial properties, and nanohydroxyapatite has good biological activity and mechanical strength, while their mixture cannot only promote bone formation but also have antibacterial properties. OBJECTIVE:To observe the effect of nanosilver and nanohydroxyapatite mixed filing on the osseointegration of immediate implants. METHODS: Nine New Zealand white rabbits were randomly divided into experimental group (n=6) and control group (n=3) after four incisors from the upper and lower jaw were extracted. The mixture of nanosilver and nanohydroxyapatite was filed into the tooth socket in the experimental group, while nanohydroxyapatite was filed into the tooth socket in the control group. Titanium screw was immediately implanted into both groups. The intact maxila and mandibular specimens were harvested at the 4th, 8th, and 12th weeks after operation. Gross observation, X-ray bone density analysis, torque test and histological observation were conducted. RESULTS AND CONCLUSION:The gray value and maximum torque value of regenerated osseous tissue at different time points in experimental group were significantly higher than those in the control group (P < 0.05). Within 12 weeks of implantation, the bone formation rate and maturity of new bone tissue were higher in the experimental group compared with the control group, and no inflammatory cel infiltration occurred. At the 4th week after implantation, there was a large amount of inflammatory cel infiltration, and few inflammatory existed at the 8th week after implantation. These results demonstrate that compared with nanohydroxyapatite alone, the mixture of nanosilver and nanohydroxyapatite shows better antibacterial effect, biocompatibility and osteoinductive ability, which may accelerate osseointegration and promote osteogenesis.

Objective To study the effect of Chinese medicinal compound recipe ( modified Fuzheng Yiliu Decoction, MFYD) on the proliferation of colon cancer HT-29 cells and on the expression of cellular apoptosis gene, Bcl-2, and tumor-inhibiting gene, P53. Methods Colon cancer HT-29 cells were divided into serum containing MFYD groups ( treated with different concentrations of serum containing MFYD) and blank serum group. Methyl thiazolyl tetrazolium ( MTT) , real time cellular analysis ( RTCA) and reverse transcription poly merase chain reaction ( RT-PCR) were used to detect the cell proliferation and mRNA expression of Bcl-2 and P53 separately. Results The proliferation of colon cancer HT-29 cell line was inhibited by MFYD ( P<0.05) in concentration-and time-dependent manner. The Bcl-2 mRNA expression was decreased and p53 mRNA was increased markedly in HT-29 cells after co-culturing with 12% or 15% volume fraction of serum containing MFYD for 48 hours, the differences being significant compared with the blank serum group ( P<0.05). Conclusion MFYD has obvious inhibitory effect on the proliferation of colon cancer HT-29 cells, and its mechanism may be related to down-regulating the expression of Bcl-2 gene and up-regulating the expression of P53 gene.

Objective To compare the clinical effect of neoadjuvant chemoradiotherapy (nCRT) and neoadjuvant chemotherapy (nCT) in the treatment of locally advanced esophageal squamous cell carcinoma.Methods The retrospective cohort study was conducted.The clinicopathological data of 156 patients with local advanced esophageal squamous cell carcinoma who were admitted to the Zhongshan Hospital of Fudan University from January 1,2010 to December 31,2015 were collected.Among 156 patients,59 undergoing nCRT were allocated into the nCRT group and 97 undergoing nCT were allocated into the nCT group.Patients in the nCRT group and nCT group respectively received 2 cycles chemotherapy by the TP regimen+40 Gy radiotherapy (2 Gy/d) and 2 cycles chemotherapy by the TP regimen.Patients were evaluated by imaging examinations after 6 weeks neoadjuvant therapy completion,and then underwent abdominal and right chest-left cervico three-incision thoracoscopic surgery.Observation indicators:(1) treatment situations;(2) postoperative pathological examination;(3) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed once every 3 months within 2 years and once every 6 months after 3 years up to January 2017.Follow-up included levels of tumor markers [carcinoembryonic antigen (CEA) and SCC-Ag],thoracic or abdominal computed tomography (CT),neck and abdominal ultrasonography and gastroscopy or PET/CT examination if necessary.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range) and comparison between groups was analyzed using the nonparametric test.Count data were analyzed using the chi-square test or Fisher exact probability.Comparison of ordinal data was done by the nonparametric test.The survival rate was calculated using the life table method and survival was analyzed by the Log-rank test.Results (1) Treatment situations:all the patients in the 2 groups were able to burden neoadjuvant therapy and thoracic esophagectomy.Six patients in the nCRT group and 15 in the nCT group had conversion to open surgery.Operation time,volume of intraoperative blood loss,cases with postoperative readmission of ICU,cases with complications,cases with perioperative death and duration of hospital stay were (201 ± 25) minutes,(137± 66)mL,5,24 (10 with pulmonary complications,8 with anastomotic leakage,3 with hoarseness,2 with cardiovascular complications and 1 with chylopleura),0,12 days (range,9-93 days) in the nCRT group and (195±20) minutes,(133±58) mL,8,30 (11 with anastomotic leakage,10 with pulmonmy complications,4 with hoarseness,2 with cardiovascular complications,1 with postoperative hemorrhage,1 with delayed gastric emptying and 1 with chylopleura),1,11 days (range,9-78 days) in the nCT group,respectively,with no statistically significant difference between the 2 groups (x2 =0.883,t =0.102,0.692,x2 =0.048,1.541,Z =0.225,P＞ 0.05).(2) Postoperative pathological examination:R0 resection rate was 96.6％ in the nCRT group and 93.8％ in the nCT group,with no statistically significant difference between the 2 groups (x2 =0.589,P＞0.05).Results of postoperative pathological examination showed that G0,G1,G2 and G3 of tumor regression grade were respectively detected in 18,16,7,18 patients in the nCRT group and 4,5,4,84 patients in the nCT group,with a statistically significant difference between the 2 groups (Z=-7.151,P＜0.05).Stage 0,Ⅰ,Ⅱ,ⅢA,Ⅲ B and ⅣA of postoperative ypTNM stage were respectively detected in 16,9,23,4,6,1 patients in the nCRT group and 4,9,37,6,34,7 in the nCT group,with a statistically significant difference between the 2 groups (Z=-4.890,P＜0.05).The down-staging was detected in 48 patients of the nCRT group and 50 patients of the nCT group,with a statistically significant difference between the 2 groups (x2=13.957,P＜0.05).(3) Follow-up and survival situations:of 156 patients,153 were followed up for 12-82 months,with a median time of 36 months.The 1-,3-,5-year overall survival rates were 88.1％,61.4％,34.9％ in the nCRT group and 81.4％,43.8％,23.1％ in the nCT group,with a statistically significant difference between the 2 groups (x2=4.336,P＜0.05).Conclusion The nCRT in the treatment of locally advanced esophageal squamous cell carcinoma can enhance postoperative pathological response rate,down-staging rate and overall survival rate compared with nCT,without increasing incidence of perioperative complications.