Objective To investigate expression of 14-3-3 sigma gene in patients with breast cancer and its clinical significance.Methods Expression of 14-3-3 sigma gene was semi-quantitated by using RT-PCR and western-blot in 40 specimens of breast cancer and 18 specimens benign breast disease tissue.Results Of 40 cases of breast cancer 35(87.5%)were negative for 14-3-3 sigma gene in RT-PCR,32(80%)were negative in Western-blot,and 31(77.5%)were negative in both RT-PCR and western-blot.Besides,the expression in 2 cases was down-regulation in both the 2 method.In 18 specimens with benign breast disease tissue the expression of 14-3-3 sigma gene was detectable,which was demonstrated by RT-PCR or western-blot.Conclusion Inactivation and down-regulation of 14-3-3 sigma gene is a frequent event in breast cancer,and it may contribute to diagnosis of breast cancer.

Objective To clone express and purify Schistosoma japonicum fructose?1 6?bisphosphate aldolase SjFBPA in E. coli and observe its expression in different developmental stages of S. japonicum. Methods FBPA gene was amplified from S. japonicum adult worm cDNA by using PCR. The amplified product was recombined into pET28a plasmid and inducibly expressed with IPTG in E. coli BL21. SDS?PAGE and Western blotting were employed to analyze and identify the recombinant protein SjFBPA rSjFBPA . Then rSjFBPA was purified by chromatographic purification and its purity was analyzed by SDS?PAGE. The protein concentration of rSjFBPA purified was measured by the BCA method. Furthermore SjFBPA mRNA was ana?lyzed in different developmental stages of S. japonicum by RT?PCR. Results SjFBPA was successfully amplified by using PCR and identified by restriction enzyme digestion and sequencing. The Western blotting analysis confirmed that the recombinant pro?tein could specifically reactive to the anti?His?tag monoclonal antibody. The concentration of the purified recombinant protein was about 4 mg/ml. The result of RT?PCR showed that SjFBPA mRNA was expressed in cercaria schistosomulum adult worm and egg of S. japonicum. Conclusion SjFBPA is successfully recombined and expressed in a prokaryotic system and SjFBPA mRNA is expressed in cercaria schistosomulum adult worm and egg of S. japonicum.

OBJECTIVE To study the genetic supports of OXA-23 in Acinetobacter baumannii isolates and investigate the relationship between imipenem resistance acquiring and use of antibiotics.METHODS Consecutive selection of the 24 highly susceptive A.baumannii clinical isolates by imipenem was carried out.Genes of carbapenemases were detected by PCR and the colonial relationship of these isolates was evaluated by ERIC-PCR.Plasmids conjugation experiments and blaOXA-23 hybridization were performed to explore the gene location of blaOXA-23.RESULTS From all 24 susceptible A.baumannii isolates 10 were selected,including 6 multi-colonial blaOXA23 harboring strains.Plasmid conjugation experiments and Southern blotting experiments demonstrated that blaOXA-23 was not associated with integrons.CONCLUSIONS BlaOXA-23 may exist in a certain subset of apparently carbapenem sensitive Acinetobacter strains.When under consecutive selective pressure,bacteria harboring blaOXA-23 become the predominant group and subsequently the antibiotics resistance properties appear.It highlights the reasonable use of this category of antibiotics.

OBJECTIVE: To investigate the effect and the possible influential mechanisms of hyperbaric oxygen (HBO) and 5-fluorouracil (5-FU) on the proliferation and metastasis in human nasopharyngeal carcinoma (NPC) CNE2Z cell line. METHOD: NPC CNE2Z cell line were divided into 4 groups randomly: group A: control group; group B: 5-FU group; group C: HBO group; group D: 5-FU and HBO group. The inhibition effect of proliferation in CNE2Z cells of all groups through 24 h, 48 h and 72 h disposal was detected by MTT reduction assay; Transwell chamber assay was performed to determine the effect of HBO and (or) 5-FU on the metastasis of the CNE2Z cells; The MMP-9, VEGF expression in CNE2Z cells of all groups were detected by SP immunocytochemical stain and observe the expressed image by micro. RESULT: There were statistical difference on the inhibition effect of proliferation in C group and A group after 48 h and 72 h disposal (P<0.01) and between A, B, C group and D group only after 48 h disposal (P<0.05); There were not statistical difference on the effect of metastasis in C group and A group (P>0.05) and between B, C group and D group (P>0.05); Average optical of the MMP-9, VEGF expression were not statistically significant difference in C group and A group (P>0.05) and between C group and D group (P>0.05). CONCLUSION Simple HBO disposal after 48 h and 72 h could inhibit the proliferation potential of NPC CNE2Z cell line, but the combination of HBO and 5-FU only after 48 h disposal could all the more inhibit the proliferation potential. Simple HBO disposal couldn't decrease the MMP-9 and VEGF high-expression and inhibit the metastasis potential in human NPC CNE2Z cell line, the combination of HBO and 5-FU disposal also couldn't further decreased the MMP-9 and VEGF high-expression and inhibited the metastasis potential.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Fluorouracil ; therapeutic use ; Humans ; Hyperbaric Oxygenation ; Matrix Metalloproteinase 9 ; metabolism ; Nasopharyngeal Neoplasms ; pathology ; therapy ; Vascular Endothelial Growth Factor A ; metabolism

Objective To evaluate the mid-to-long term therapeutic effect of endoscopic pneumatic dilation for achalasia of cardia (AC).Methods Endoscopic pneumatic dilation was used in 45 AC patients, with follow-up of 2-12 years. Eckardt score and Stooler grading were used before and after the operation for evaluation of curative effect of dilation. Results The operation success rate was 97. 8%( 44/45) and the effective remission rate was 93. 2%( 41/44 ). No massive hemorrhage, perforation or other serious complications occurred.The longest follow-up time was up to 144 months.Ten cases were followed up for over 60 months. Patients′symptoms relieved significantly (P＜0. 01). Eckardt scores in 24 months and 60 months after operation significantly decreased compared with those before the operation ( P＜0. 01). But Eckardt score in 60 months was higher than that in 24 months ( P＜0. 01). The length of the disease history was positively related to post-operative scores, and negatively related to efficacy ( P＜0. 01). Conclusion Endoscopic balloon dilation is a satisfactory therapy to AC with good efficacy and safety.

Objective:To intestigate the clinical efficacy between modified Overlap anastomosis and traditional auxiliary incision anastomosis in laparoscopic total gastrectomy.Methods:The retrospective cohort study was conducted. The clinicopathological data of 115 patients with gastric cancer who were admitted to the Second Affiliated Hospital of Fujian Medical University from January 2016 to December 2018 were collected. There were 62 males and 53 females, aged from 27 to 83 years, with a median age of 62 years. Of 115 patients, 51 patients undergoing totally laparoscopic total gastrectomy with modified Overlap anastomosis using linear stapler were divided into modified Overlap group and 64 patients undergoing laparoscopic assisted total gastrectomy with traditional auxiliary incision anastomosis using circular stapler were divided into traditional assisted group. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) anastomotic complications; (4) follow-up. Follow-up using outpatient examination or telephone interview was conducted to detected tumor recurrence and survival of patients up to December 2019. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Count data were represented as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Comparison of ranked data was analyzed using the rank sum test. Results:(1) Surgical situations: the operation time, time of esophagojejunostomy, volume of intraoperative blood loss, the number of lymph node dissected, length of proximal incisional margin and length of auxiliary incision of the modified Overlap group were (234.0±11.0)minutes, (29.4±2.1)minutes, (53±14)mL, 42±13, (2.0±0.3)cm and (5.1±0.4)cm, respectively. The above indicators of the traditional assisted group were (231.0±11.0)minutes, (29.2±2.2)minutes, (50±13)mL, 40±10, (2.2±0.4)cm and (8.2±0.4)cm, respectively. There was significant difference in the length of auxiliary incision between the two groups ( t=-43.098, P<0.05), and there was no significant difference in the operation time, time of esophagojejunostomy, volume of intraoperative blood loss, the number of lymph node dissected, length of proximal incisional margin between the two groups ( t=1.168, 0.460, 0.990, 1.127, -1.926, P>0.05). (2) Postoperative situations: cases with mild, moderate, severe pain (postoperative pain degree), time to first flatus, time to initial fluid diet intake, duration of postoperative hospital stay of the modified Overlap group were 40, 9, 2, (2.9±1.0)days, (4.8±2.2)days, (11.7±2.8)days, respectively. The above indicators of the traditional assisted group were 31, 27, 6, (3.9±1.4)days, (6.5±2.5)days, (13.0±3.1)days, respectively. There were significant differences in the above indicators between the two groups ( Z=-3.217, t= -4.344, -3.888, -2.261, P<0.05). (3) Anastomotic complications: cases with anastomotic leakage, cases with anastomotic bleeding, cases with anastomotic stenosis of the modified Overlap group were 1, 1, 0, respectively. The above indicators of the traditional assisted group were all 1. There was no significant difference in the above indicators between the two groups ( P>0.05). Cases with anastomotic leakage were cured after the treatment of enteral nutritional support through nasogastric catheterization, which were confirmed by gastroenterography. Cases with anastomotic bleeding were improved by active hemostatic therapy. Cases with anastomotic stenosis were improved after the symptomatic treatment of anti-inflammatory and anti-swelling. (4) Follow-up: 109 of the 115 patients were followed up. Forty-eight of 51 patients in the modified Overlap group were followed up for 15.0-45.0 months, with a median follow-up time of 33.5 months. Sixty-one of 64 patients in the traditional assisted group were followed up for 16.0-46.0 months, with a median follow-up time of 27.0 months. There was no tumor recurrence in the modified Overlap group. One patient in the traditional assisted group had tumor recurrence with liver metastasis and survived with tumor. There was no significant difference in tumor recurrence rate between the two groups ( P>0.05). There was no patient died during the follow-up. Conclusion:Compared with traditional auxiliary incision anastomosis, patients undergoing total laparoscopic total gastrectomy with modified Overlap anastomosis have small incision, good postoperative recovery.

Objective To determine the seasonal fluctuation and population distribution of Aedes albopictus in Jiading District， and provide scientific evidence for the prevention and control of dengue fever and other Aedes-borne diseases. Methods In 2020， the mosq-ovitrap method and mosq-ovitrap index （MOI） were used to monitor and evaluate the density of Aedes albopictus in Jiading District. Spatial and temporal distribution of Aedes albopictus was determined. Chi-square test was used for statistical analysis. Results In 2020， the annual average MOI was determined to be 4.10， which was under safety threshold. The seasonal fluctuations showed a unimodal distribution， which peaked in July. The fluctuation trend in urban area was similar to the overall trend， while that in the non-urban area showed a bimodal distribution with peaks in June and August. The density of Aedes albopictus at different monitoring sites varied widely，with the highest MOI （6.64） at Anting town and the lowest MOI （2.09） at Huating town. The distribution of Aedes albopictus in different habitats also varied widely； the highest density was observed in environments as waste collection stations and construction sites， with the highest MOI 33.33 in waste collection stations in peak season. The MOI value of Aedes albopictus in residential areas was significantly higher than that in non-residential areas （χ² = 6.082， P = 0.014）. Conclusion Aedes albopictus is quite common in Jiading District. In certain areas， Aedes density may exceed the safety threshold from May to September. More targeted mosquito control measures should be implemented in waste collection stations， construction sites and residential areas.

ObjectiveTo investigate the impact of icariin （ICA） on autophagy in glucocorticoid-induced bone microvascular endothelial cells （BMECs） mediated by the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway. MethodBMECs were isolated and cultured from femoral heads obtained during total hip arthroplasty and identified using immunofluorescence staining. The experimental cells were divided into four groups： A control group， a glucocorticoid group （100 mg·L^-1 hydrocortisone）， an ICA group （100 mg·L^-1 hydrocortisone+6.7×10^-3 mg·L^-1 ICA）， and a Rapamycin group （100 mg·L^-1 hydrocortisone+6.7×10^-3 mg·L^-1 ICA+1 mg·L^-1 rapamycin）. Autophagy in BMECs was induced using 100 mg·L^-1 hydrocortisone. LC3 fluorescence staining was used to observe the peak of autophagy at different time points. Western blot analysis was employed to analyze the expression of autophagy-related proteins and PI3K/Akt/mTOR pathway proteins in each group. Electron microscopy was used to observe autophagosomes and autolysosomes in the cells. ResultHydrocortisone at 100 mg·L^-1 induced autophagy in BMECs， reaching a peak at around 5 hours， which then declined with further intervention. Compared to the control group， the glucocorticoid group showed cell membrane damage， disordered organelle arrangement， and a large number of autophagosomes and autolysosomes. Compared to the glucocorticoid group， the ICA group had more intact cell membranes， sparser organelle arrangement， and fewer autophagosomes and autolysosomes. Compared to the ICA group， the Rapamycin group showed cell membrane damage， disordered organelle arrangement， and more autophagosomes and autolysosomes. Compared to the control group， the glucocorticoid group had significantly increased expression of light chain 3B （LC3B）， Atg4B， and p62 （P<0.01）. Compared to the glucocorticoid group， the ICA group showed significantly decreased expression of LC3B， Atg4B， p62， and Beclin-1 （P<0.01）. Compared to the ICA group， the Rapamycin group had significantly increased expression of Atg4B and p62 （P<0.01）. Compared to the control group， the glucocorticoid group had significantly decreased expression of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR （P<0.01）. Compared to the glucocorticoid group， the ICA group showed significantly increased expression of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR （P<0.01）. Compared to the ICA group， the Rapamycin group had significantly decreased expression of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR （P<0.01）. Ubiquitination levels were significantly decreased in the glucocorticoid group compared to the control group （P<0.01）. Compared to the glucocorticoid group， ubiquitination levels were significantly increased in the ICA group （P<0.01）， and significantly decreased in the Rapamycin group compared to the ICA group （P<0.01）. ConclusionThe glucocorticoid-induced autophagy in BMECs is time-dependent. ICA inhibits glucocorticoid-induced autophagy in BMECs， and this effect may be related to the regulation of the PI3K/Akt/mTOR pathway.