Hepatocellular carcinoma （HCC） with biliary duct tumor thrombus （BDTT） is currently not common in clinical practice and is easily misdiagnosed， and previously， it was often considered an advanced stage of the disease with a poor prognosis， making its treatment challenging. However， in-depth studies in recent years have gradually deepened our understanding of this disease， leading to significant changes in diagnostic and treatment concepts. Currently， comprehensive treatment， mainly surgery， is used for treatment， but there is still controversy over the selection of clinical treatment strategies. This article provides a detailed discussion on surgical methods and prognosis， in order to provide a reference for clinical treatment options.

  Objective  To promote the development of high-quality health standards in China and summarize the characteristics of global health standards management systems and development processes.  Methods  We conducted a systematic search of relevant databases and the websites of standardization institutions. A descriptive method was used to analyze the basic characteristics of the standard management systems and development processes.  Results  The majority of the investigated countries and organizations primarily implement voluntary standards, with the exception of Chinese and the European Committee for Standardization that enforce compulsory standards. All countries and organizations have health standards at the national, group, and enterprise levels. In China and Japan, standards authorities are subordinate to the government, while in other countries or organizations, they operate as independent civil societies. The development process for health standards generally involves five stages: "Project/Proposal → Draft → Enquiry → Review → Publication". However, the specific steps in the development of health standards vary across different countries and organizations. Furthermore, China ranks high in the number of both standards and health standards issued, but the proportion of health standards is only 2.06%.  Conclusions  There are significant differences between China and other countries in terms of the management system and development process of health standards. To promote the management and development of high-quality health standards in China, we recommend promoting the coordination mechanism for standard development, increasing efforts to publicize standards and evaluate the implementation effect, strengthening the construction of personnel for the development of health standards, optimizing the efficiency of standard development and improving the internationalization level of health standards.

The paper-based microfluidic chip,also known as a paper chip,uses paper as the substrate on which the sample processing,biochemical reactions and assay processes are performed.Com-pared to other microfluidic chips,the paper chip has the advantages of widely availability of raw materials,lower cost,and easier disposal after use and ease of operation,making it more suitable for rapid on-site testing.This study systematically summarizes the production technology of paper chip and its research progress in animal pathogen detection,analyzes the existing problems of pa-per chip technology,and looks forward to the future research direction,so as to provide reference for the improvement of paper chip technology and its popularization and application.

Objective: To explore the influence of neonatal intensive care unit (NICU) environmental factors on physiological function of premature infants, and propose improvement plan. Methods: A total of 40 cases of low body mass premature infants from June to December in 2017 in Chongqing Center for Women and Children were selected as the research objects, and 8:00-9:00 of working day was set as the daily period, 10:00-11:00 as the quiet period. Light, sound, touch and other environmental factors in NICU were detected quantitatively, basic vital signs, activity, stress hormones and other changes were recorded, and the impact of environmental factors on the children was analyzed. Results: Touch times in daily and quiet periods were (5.02±0.54) times/h and (4.56±0.55) times/h, with no significant difference (P>0.05). The same time as the next two days, the activity of premature infants in quiet period was (8.26±3.10), which was lower than that in daily period (17.52±5.78), and the difference was statistically significant (t value was 8.967, P<0.01). The sleep time of premature infants during quiet period (23.24±8.38) was significantly higher than that of daily period (4.80 ±5.39), the difference was statistically significant (t value was-11.679, P<0.01). The levels of cortisol, epinephrine and norepinephrine in premature infants were (61.53±13.47), (15.91±3.94), (49.22±15.15) μg/L in quiet period, which were significantly lower than those in daily period (74.52±20.98), (23.40±11.66), (80.32±32.43) μg/L (t value was 3.295, 3.848, 5.502, P<0.01). The heart rate and systolic pressure of premature infants in quiet period (130.44±8.06) beats/min, (64.05±10.40) mmHg (1 mmHg=0.133 kPa) were significantly lower than those in daily period (145.21±10.52) beats/min, (72.85±13.36) mmHg, and the difference was statistically significant (t value was 3.340, 2.166, P<0.01 or 0.05). Different time periods on the same day, the activity of premature infants in quiet period was (9.87±3.31), which was lower than that in daily period (19.82±5.57), and the difference was statistically significant (t value was 9.761, P<0.01). The sleep time of premature infants during quiet period (25.75±9.07) was significantly higher than that of daily period (4.70±3.89), the difference was statistically significant (t value was-13.457, P<0.01). The levels of cortisol, epinephrine and norepinephrine in premature infants were (59.42±11.95), (15.78±1.45), (50.90±14.73) μg/L in quiet period, which were significantly lower than those in daily period (71.50±20.56), (25.62±11.95), (79.87±29.91) μg/L (t value was 3.212, 5.169, 3.585, P<0.01). The heart rate and systolic pressure of premature infants in quiet period (136.02±11.22) beats/min, (65.35±9.56) mmHg were significantly lower than those in daily period (144.10±9.18) beats/min, (73.47±12.92) mmHg, and the difference was statistically significant (t value was 3.533, 2.008, P<0.01 or 0.05). Conclusions Children with NICU are generally in high noise and strong light environment, these factors will cause adverse physiological reactions to a certain extent.

Objective To investigate the application value of choledochoscopy and imaging examination in the diagnosis and treatment of residual bile duct stones.Methods The retrospective descriptive study was conducted.The clinical data of 618 patients who underwent choledochoscopy and imaging examination after operation of hepatolithiasis in the First Affiliated Hospital of Zhengzhou University between April 2014 and September 2018 were collected,including 300 males and 318 females,aged from 19 to 89 years,with an average age of (58 ± 12)years.Observation indicators:(1) situations of residual bile duct stones diagnosed by choledochoscopy and imaging examination;(2) stone extraction situations of patients with hepatolithiasis.Measurement data with normal distribution were represented as Mean±SD,and measurement data with skewed distribution were described as M (range).Count data were represented as absolute number or percentage,and analyzed using the chi-square test or Fisher exact propability.Results (1) Situations of residual bile duct stones diagnosed by choledochoscopy and imaging examination:all the 618 patients underwent choledochoscopy,and 505 of them underwent the imaging examination before choledochoscopy.① Of patients undergoing single imaging examination,72 received ultrasonography,with false-negative rate of 29.17％ (21/72);37 received CT examination,with false-negative rate of 10.81％(4/37);33 received T-tube cholangiography,with false-negative rate of 39.39％ (13/33).② Of patients undergoing combined two imaging examinations,61 received ultrasonography + CT,with false-negative rate of 8.20％ (5/61);129 received ultrasonography + T-tube cholangiography,with false-negative rate of 12.40％ (16/129);52 received CT + T-tube cholangiography,with false-negative rate of 5.77％(3/52).③ There were 121 receiving ultrasound+CT+T-tube cholangiography,with false-negative rate of 7.44％ (9/121).There were statistically significant differences in the false-negative rates of combined two or three examinations of ultrasound + CT+ T-tube cholangiography and single imaging examination (x2=40.83,P＜0.05).The further analysis showed a statistically significant difference among the single imaging examination (x2=7.70,P＜0.05).There was no statistically significant difference among the combined two of imaging examinations (x2=2.10,P＞0.05).There were statistically significant differences in the combined three examinations of ultrasound +CT+T-tube cholangiography and ultrasound and T-tube cholangiography examination respectively (x2=16.23,21.62,P＜0.05).There was no statistically significant difference in the combined three of imaging examinations and CT examination and combination of CT+T-tube cholangiography respectively (P＞ 0.05).There was no statistically significant difference in the combined three of imaging examinations and combination of ultrasound+CT examinations and combination of ultrasound+T-tube cholangiography (x2=0.33,1.71,P＞0.05).Seventy-one patients without residual bile duct stone by preoperative imaging examination were detected residual bile duct stones by intraoperative choledochoscopy,and residual bile duct stones of 36,31 and 4 patients are respectively distributed around the distal common bile duct,small intrahepatic bile duct,left and right hepatic ducts,common hepatic duct and remaining common bile duct.(2) Stone extraction situations of patients with hepatolithiasis:of 618 patients,cases with 1,2,3,4,5,6,7,8,9 and 10 times of residual bile duct stones clearance were respectively 392,116,48,39,9,6,3,2,2 and 1.Residual bile duct stones clearance frequency of patients was an average of 1.73 times.There were 63.43％(392/618) and 96.28％(595/618) of patients had stone clearance with once and ≤ 4 times of stone extraction,respectively.Conclusion The negative results of preoperative imaging examinations cannot be as standards of bile duct stone clearance before choledochoscopy,and the best choice is to detect whether there are residual bile duct stones and remove the stones combined with choledochoscopy.

Objective: To explore the clinical efficacy of S-1 single agent adjuvant chemotherapy for the patients undergoing radical resection of extrahepatic biliary carcinoma. Methods: The clinical data of 108 patients with extrahepatic biliary carcinoma receiving radical resection who were admitted from January 2014 to June 2017 were retrospectively analyzed. There were 62 males(57.4%)and 46 females(42.6%),with a median age of 59 years (range:26 to 79 years),10 cases(9.3%) in stage Ⅱ,85 cases(78.7%) in stage Ⅲ, and 13 cases (12.0%) in stage Ⅳ, 40 cases(37.0%) of hilar cholangiocarcinoma, 8 cases(7.4%) of middle cholangiocarcinoma, 25 cases (23.2%) of distal cholangiocarcinoma, 35 cases(32.4%) of gallbladder carcinoma.After radical resection of extrahepatic biliary carcinoma, 49 patients receiving S-1 single agent chemotherapy and 59 patients receiving non-special treatment were divided into the chemotherapy group and the operation group,respectively. All the dates of the patients were followed up and collected with the overall survival time,tumor-free survival time,1,2 and 3-year survival rate after operation,and the rate of major toxic reaction during chemotherapy of the chemotherapy group. Survival curve was drawn by the Kaplan-Meier method, and survival analysis was done using the Log-rank test. Results: There were no significant differences in the general date of two groups(sex, age, tumor size, tumor site, TNM stages, degree of differentiation). The median overall survival time and the median tumor-free survival time in the chemotherapy group were 27 months and 21 months,respectively,and in the operation group were 21 months and 17 months,respectively. There were differences between the two groups in the overall survival rates(χ²=3.967,P<0.05) and the 2 and 3-year survival rate(63.3%,36.6%;41.6%,20.4%;χ²=4.510,P<0.05;χ²=6.143,P<0.05),but the 1-year overall survival rate (83.4%,79.7%)was not statistically significant(χ²=0.286,P>0.05). There were no significant differences in the tumor-free survival time,1,2 and 3-year tumor-free survival rate(77.6%,41.4%,33.1%;62.7%,30.9%,21.2%)between the two groups(χ²=0.876,P>0.05;χ²=0.252,P>0.05;χ²=1.571,P>0.05;χ²=3.323,P>0.05,respectively). The main toxic reaction during chemotherapy were dyspepsia(28.6%, 14/49), anemia(26.5%, 13/49), and leukopenia(22.5%, 11/49), all of which were mild. Conclusion S-1 single agent chemotherapy after radical reseetion of extrahepatic biliary carcinoma could effectly improve the survival of patients and all of the main toxic reaction during chemotherapy were mild.

Objective To explore the expression of calreticulin (CRT) in gallbladder cancer tissue and its effect on the biological behavior in gallbladder cancer GBC-SD cells.Methods Immunohistochemistry and RT-qPCR were applied to detect the expression of CRT.Small interfering RNA was transfected into gallbladder cancer GBC-SD cells and Western blotting were used to detect the expression of CRT.The proliferation was determined by using cell counting kit-8 (CCK-8) and clone assays.Flow cytometry were applied to detect the apoptosis and cell cycle.Migration was detected by wound healing and transwell assays,respectively.The expression of p-Akt and MMP-9 were detected by using Western blotting.Results Expression of CRT in gallbladder cancer tissues is higher than adjacent cancer tissues and chronic cholecystitis tissues(t =5.571,P ＜ 0.05).The relative growth rate in the siCRT-1,siCRT-2 experimental group for 24 hours,48 hourrs were 71.5％ ±6.3％,79.5％ ±2.7％;62.6％ ± 8.8％,55.6％ ±2.6％,respectively.The apoptosis rate in the blank group,the negative control group,siCRT-1 and siCRT-2 group were 3.0％ ± 1.8％,4.7％ ± 1.3％,13.6％ ± 1.0％,20.0％ ± 4.0％,respectively.Wound healing assays showed that the wound closure ratio in the blank group,negative control group,siCRT-1 and siCRT-2 group were(0.67 ±0.02),(0.58 ±0.02),(0.22 ±0.01),(0.37 ±0.04),respectively.Transwell experiments showed that the numbers of migration of GBC-SD cells in the blank group,negative control group,siCRT-1 and siCRT-2 group were (302 ± 11),(297 ± 15),(178 ± 10),(165 ± 12),respectively,compared with the blank group and the negative control group,the relative growth rate for 24 hours and 48 hours was significantly lower,the apoptosis rate was higher,the numbers of migration was lower (F =29.310,118.618,69.651,144.515,190.145,P ＜ 0.05).Compared with the blank group and the negative control group,the expression of p-Akt and MMP-9 decreased after down-regulating the expression of CRT.Conclusions The expression of CRT in gallbladder cancer tissue was higher.CRT downregulation mediated changes of biological behaviors in gallbladder cancer may be associated with p-Akt/MMP-9 signal pathway.

Objective: To investigate the effects of Kindlin-2 on malignant phenotypes of human gallbladder cancer cells and discuss the mechanisms. Methods: The expression level of Kindlin-2 in 30 cases of gallbladder cancer tissues and adjacent non-tumoral tissues collected from the First Affiliated Hospital of Zhengzhou University between September 2012 and May 2013 was assessed by real-time PCR and immunohistochemistry.Lentivirus-mediated Kindlin-2 overexpression was used in gallbladder cancer cell lines GBC-SD and SGC-996.Transwell assay and adhesion assay were investigated to explore the functional role of Kindlin-2 on gallbladder cancer cells.Western Blot was used to test the protein change of epithelial-mesenchymal transition(EMT) characteristics. The t-test was used to analyzed results. Results: The RNA and protein levels of Kindlin-2 in gallbladder cancer tissues were higher than in the non-tumoral tissues (t=4.372, P=0.001; t=7.477, P=0.000). The expression level of Kindlin-2 in gallbladder cancer tissues was correlated with Nevin stage(χ²=5.932, P=0.035). Compared with control groups, the cell-matrix adhesion ability of GBC-SD and SGC-996 with Kindlin-2 overexpression was obviously promoted(1.66±0.03 vs. 1.07±0.22, t=2.710, P=0.041; 2.66±0.24 vs. 1.03±0.02, t=6.610, P=0.020). The number of GBC-SD and SGC-996 cells with Kindlin-2 overexpression passing through the Transwell chamber matrix increased significantly compared with the control groups(116.1±13.9 vs. 54.7±8.4, t=3.781, P=0.019; 136.3±7.5 vs. 64.3±6.4, t=7.302, P=0.002). The wound healing rate of GBC-SD with Kindlin-2 overexpression at 12-hour and 24-hour was higher than that of the group ((42.9±2.2)% vs. (29.7±1.7)%, t=4.690, P=0.009; (65.0±2.4)% vs.(40.4±2.0)%, t=7.945, P=0.001). The wound healing rate of SGC-996 with Kindlin-2 overexpression at 12-hour and 24-hour was also higher than that of the group ((32.9±1.3)% vs. (24.1±1.5)%, t=4.518, P=0.011; (51.3±1.1)% vs. (39.2±1.1)%, t=8.001, P=0.001). The characteristics of EMT were induced in gallbladder cancer cells with Kindlin-2 overexpression, including the up-regulation of N-cadherin, Vemintin and the down-regulation of E-cadherin. Conclusion The expression of Kindlin-2 is up-regulated in gallbladder cancer tissues and Kindlin-2 promoted the malignant phenotypes of gallbladder cancer cells partially by epithelial-mesenchymal transition.

Objective To investigate the efficacy and safety of capecitabine in the treatment of colorectal cancer. Methods Totally 160 elderly patients with stageⅣcolorectal cancer were enrolled in this study. After first-line combined chemotherapy,80 patients were treated with capecitabine monotherapy(maintenance group)and another 80 cases were not(control group). The survival rate was analyzed by Kaplan-Meier curve and the efficiency and incidence of adverse events were compared. Results (1) The Kaplan-Meier curve suggested that the difference between two groups was statistically significant(P<0.05).(2)The response rate of maintenance group was significantly higher than that of control group (P < 0.05). (3)The incidence of adverse events during capecitabine monotherapy was lower than that during combined chemotherapy(P < 0.05).(4)The incidence of adverse reactions during capecitabine monotherapy was similar to that of control group(P > 0.05). Conclusion Capecitabine monotherapy in patients with stage Ⅳ colorectal cancer after combined chemotherapy has a longer median PFS than those without maintenance but similar adverse reactions ,which was worthy of clinical promotion.

Objective To study the expression and the clinical significance of hypoxia-induced factor-1α (HIF-1α) in gallbladder cancer tissues,and the role and mechanism of HIF-1α in metformin-suppressed metastasis in gallbladder carcinoma GBC-SD cells.Methods 24 specimens of gallbladder cancer tissues and 5 specimens of chronic cholecystitis were collected from the First Affiliated Hospital of Zhengzhou University between June 2016 and February 2017.Immunohistochemistry and qPCR were used to detect the expression of HIF-1α in gallbladder cancer tissues,in adjacent non-cancer tissues and in chronic cholecystitis,and the clinical significance was analyzed.The model of metastasis was induced by hypoxia;the wound healing assay and the Transwell assay were used to detect the ability of cell metastasis;the expressions of HIF-1α and VEGF in gallbladder carcinoma GBC-SD cells were detected by western blotting assay and immunofiuorescence.Results The expression of HIF-1α in gallbladder cancer tissues was higher than the adjacent non-cancer tissues and in chronic cholecystitis.The expression of HIF-1α was correlated with lymph node metastasis and TNM staging in gallbladder cancer tissues (P ＜ 0.05).The wound healing rate after 48 h in the negative control group and in the treatment with hypoxia group (1％ O2) in GBC-SD cells were (46.5 ± 4.8) ％ and (67.3 ± 4.0) ％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group and in the treatment with hypoxia group GBC-SD cells were (147.4 ± 11.7) and (234.4 ± 17.7),respectively.When compared with the negative control group,treatment with hypoxia significantly increased the ability of metastasis and up-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).The wound healing rate after 48 h in the negative control group,the metformin group,the hypoxia group and the metformin and hypoxia group in GBC-SD cells were (40.6 ± 7.1) ％,(16.4 ± 9.4) ％,(69.5 ± 4.0) ％ and (22.4 ± 7.4) ％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group,the metformin group,the hypoxia group and the metformin and hypoxia group in GBC-SD cells were (148.4 ± 6.9),(90.0 ± 8.4),(185.8 ± 10.2) and (113.4± 8.6),respectively.When comparcd with the hypoxia group,treatment with metformin and hypoxia significantly decreased the ability of metastasis and down-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).The wound healing rate after 48 h in the negative control group,the 2MeoE2 group,the hypoxia group,the 2MeoE2 and hypoxia group in GBC-SD cells were (43.4 ±4.4)％,(25.9 ±9.0)％,(63.3 ±2.2)％,(46.2 ±4.5)％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group,the 2MeoE2 group,the hypoxia group,the 2MeoE2 and hypoxia group in GBC-SD cells were (144.2 ± 12.6),(80.2 ±7.7),(203.8 ±7.0),(124.0 ± 5.2),respectively.When compared with the hypoxia group,treatment with HIF-1α inhibitor 2MeoE2 and hypoxia significantly decreased the ability of metastasis and down-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).Conclusions The expression of HIF-1 α was correlated with lymph node metastasis and TNM staging in gallbladder cancer tissues.Treatment with hypoxia significantly increased the expression of HIF-1α and VEGF and promoted metastasis of GBC-SD cells,while treatment with metformin decreased the ability of metastasis induced by hypoxia via inhibiting the HIF-1o/VEGF pathway in GBC-SD cells.