Objective Blocking the expression of Survivin with siRNA, and the effects of suppling the proliferation of PC-2 cell and inducing its apoptosis were investigated. Methods Constructed the siRNA against Survivin plasmid expression vector and transfected it into PC-2 cell with lipofectamineTM 2000, the changes of Survivin expression were detected by semi-quantitive RT-PCR and immunohistochemical method, The effect of suppressing the proliferation of PC-2 cell was detected by the method of MTT; the effect of inducing PC-2 cell apoptosis was detected by flow cytometry. Results The sequence specific siRNA can effectively block the expression of Survivin both at themRNA and protein levels, the expression inhibition ratio was 81.25% at mRNA level and 74. 24% at protein level;blocking the expression of Survivin can suppress the proliferation of PC-2 cell significantly, 24, 48 hours after the cell was reseeded, the proliferation inhibition ratio was 28. 00% and 33. 38% respectively; 24, 48 hours after the transfection, 8.46 % and 7.53 % cells were induced to apoptosis respectively. Conclusion The siRNA against Survivin plasmid expression vector constructed in the study can blocking the expression of Survivin in PC-2 cell effectively and specifically; blocking the expression of Survivin can signiilcantly suppress the proliferation of PC-2 cell and induce a certain degree cells apoptosis; RNAi against Survivin is of a certain value in the gene therapy of pancreatic cancer.

Objective:To observe the effects of continuous light exposure on skeletal muscle fiber type transformation and lipid metabolism, and to explore its internal relationship.Methods:Mice were randomly divided into normal light group and 24-hour continuous light group by random number table. The serum and skeletal muscle lipid content and urine 6-sulfatoxymelatonin(6-SML)level were detected by ELISA. The expression of circadian clock and lipid metabolism related genes mRNA were observed by realtime PCR. The muscle fiber type and lipid deposition were evaluated by tissue immunofluorescence as well as oil red O staining.Results:Compared with the normal light group, the level of 6-SML in urine at night decreased( P<0.05), and the expression level and rhythm of brain and muscle ARNT-like protein 1(Bmal1), circadian locomotor output cycles protein kaput(Clock), and period 2(Per2)mRNA in the skeletal muscle changed in continuous light group. In addition, the body weight, blood lipid, free fatty acid, and triglyceride contents of skeletal muscle in continuous light group increased significantly( P<0.05 or P<0.01), the expression of carnitine palmitoyltransferase 1b (Cpt1b)mRNA, the key enzyme of fatty acid oxidation, decreased significantly( P<0.05), while the expression of stearoyl-CoA desaturase(Scd1)mRNA, a lipid synthesis related gene, increased significantly( P<0.01). Further immunofluorescence analysis showed that the proportion of slow muscle fibers decreased and that of fast muscle fibers increased in continuous light group(both P<0.05). Conclusion:The process of ectopic deposition of lipid in skeletal muscle in mice induced by continuous light exposure may be related to the remodeling of skeletal muscle fibers.

Objective To investigate the epidemiological characteristics of multiple-injuries of bone and joint in the belief that a better knowledge of such injuries may help their prevention and treatment. Methods A retrospective study was done on the data of 346 patients with multiple-injuries of bone and joint who had been ad- mitted to our department from January 2001 to December 2004. On the basis of CAI's classification, the following data were statistically analyzed: gender, age, cause of injury, injured part, number of injured parts, associated injuries and mortality. Results Of the 346 injured patients, there were 278 males and 68 females, with an av- erage age of 32.8 years (9months to 89 years). Two hundred and twenty-six cases resulted from traffic accidents, 65 from crush by a heavy object, and 52 from falling. There were 159 fractures of shaft of tibia and fibula, 96 fractures of femoral shaft, 87 fractures of shaft of ulna and radius, 58 fractures of ankle and foot, 57 chest injuries, 50 knee injuries, 50 hip injuries, 49 injuries at the pelvis region, 46 wrist and hand injuries, 36 injuries of shoulder, 36 skull fractures, 33 fractures of humeral shaft, 23 spinal fractures, and 17 elbow injuries. Two hundred and forty-two patients had two parts injured, 83 had three parts, 20 had four parts, and one had six parts. The average number of injured parts was 2.3. Two hundred and five patients suffered from close injuries, and 141 from open ones. The associated injuries included skull and brain injury in 51 cases, chest injury in 23, abdomen injury in five, urine system injury in three, nerve and vessel injury in 21, shock in 78, and fat embolism in six. Five patients died. Conclusions Male young people tend to be the majority of victims of multiple-injuries of bone and joint. Traffic accidents result in most of such injuries. Since multiple-injuries mostly involve lower limbs, they are easy to diagnose while the associated close injuries involving brain, chest, abdomen and pelvic are likely to be overlooked or misdiagnosed. Strengthening safety education and technical training of first aid is important to im- provement of treatment and to decrease of disability rate and mortality.

Objective To explore the value of elastography score and strain rate ratio in the diagnosis of small breast malignant focus. Methods We retrospectively analyzed 22 patients with breast small malignant foci less than 10 mm. Ultrasound characteristics were summed up in breast small cancer. Results On elastogram, 2 patients scored 3, 14 scored 4 and 6 scored 5.The average strain rate ratio of all foci was 4.76, and there was correlation between it and elastography scores. Conclusion Ultrasonic elastography has important value in the diagnosis of breast small cancer.

Objective To use Excel worksheet function to achieve statistical analysis of the fourfold table qualitative data,and to improve knowledge of the.majority of researchers on the statistical capabilities of Excel.Methods The result of actual research published in Chinese Journal of Endemiology was the data source,and statistical analysis results of fourfold table data from Excel and from SAS software were contrasted.Results The worksheet function of Excel could realize the analysis fourfold table data chi-square test,obtained an accurate P values,but not just P ＞ 0.05 or P ＜ 0.05.The results of chi-square test of Excel were the same as the results from SAS software,but Excel could not obtain probability value of fisher exact probability.Conclusion As a dependent statistical analysis software,Excel is an easy to learn,easy to use,efficient and a good helper.In the absence of professional statistical software,Excel can be used to achieve data statistical analysis.

Objective:To investigate the application value of laparoscopic duodenum-preserving pancreatic head resection (LDPPHR).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 25 patients undergoing LDPPHR in the Second Hospital of Hebei Medical University from November 2016 to November 2020 were collected. There were 7 males and 18 females, aged from 14 to 66 years, with a median age of 29 years. All the 25 patients underwent LDPPHR. Observation indicators: (1) surgical situations; (2) postoperative histopathological examination; (3) follow-up. Follow-up was conducted using outpatient examination or telephone interview to detect the recovery of patients up to March 2021. Measurement data with skewed distribution were represented as M (range) and count data were descripted as absolute numbers. Results:(1) Surgical situations: all the 25 patients underwent LDPPHR successfully, including 23 cases undergoing total pancreatic head resection and 2 cases undergoing subtotal pancreatic head resection. The operation time and volume of intraoperative blood loss of 25 patients were 310 minutes (range, 207 to 540 minutes) and 200 mL (range, 50 to 800 mL), respectively. Of the 25 patients, 1 case was infused with 4 U of red blood cells and 400 mL of plasma, 1 case was infused with 500 mL of plasma, 1 case was infused with 600 mL of plasma and the remaining 22 cases were not infused with red blood cells or plasma. Of the 25 patients, 3 cases with pancreatic fistula of class B were discharged after drainage, 4 cases had biliary fistula including 2 cases undergoing symptoms disappeared after implantation of common bile duct stent by endoscopic retrograde cholangiopancreatography, 1 case recovering well with drainage, 1 case with postoperative perihepatic effusion undergoing symptoms disappeared after the treatment of drainage and common bile duct stent implantation, and the remaining 18 cases had no complications. The duration of postoperative hospital stay was 17 days (range, 9 to 27 days) of the 25 patients. (2) Postoperative histopathological examination: the tumor volume of the 25 patients was 6.0 cm×5.0 cm×2.0 cm (range, 1.0 cm×2.0 cm×1.5 cm to 10.0 cm×9.0 cm×8.0 cm). Results of the postoperative histopathological examination showed that there were 12 cases with pancreatic solid pseudopaillary neoplasm, 4 cases with intraductal papillary mucinous neoplasm, 3 cases with serous cystadenoma, 2 cases with mucinous cystadenoma, 1 case with neuroendocrine neoplasm, 1 case with pancreatic true cyst, 1 case with cholesterol crystals combined with calcification in the center of pancreatic nodules and 1 case with cavernous hemangioma of pancreas. (3) Follow-up: all the 25 patients were followed up for 4 months to 48 months, with a median follow-up time of 27 months. During the follow-up, 1 case of the 25 patients with postoperative diabetes controlled blood glucose in the normal range after regular injection of insulin, 1 case with fatty diarrhea had symptoms improved after oral supplement of pancreatic enzyme preparation, 1 case with preoperative intermittent dizziness, weakness of both lower limbs and hypoglycemia had the level of blood glucose returned to normal without any special treatment after operation, and the remaining 23 cases had no metabolic complications. None of the 25 patients had tumor malignant transformation, recurrence or death. No long-term complications such as delayed gastric emptying, bile duct stones or stricture occurred to the 25 patients.Conclusion:LDPPHR is safe and feasible for the treatment of benign or low-grade malignant tumors of the pancreatic head, with the advantage of preserving the integrity of digestive tract.

OBJECTIVETo investigate the effect of a sequence-specific small interfering RNA (siRNA) in suppressing survivin expression and cell proliferation and inducing apoptosis of PC-2 cells.

METHODSThe plasmid expression vector of siRNA targeted against survivin was constructed and transfected into PC-2 cells with Lipofectamine 2000. The changes of survivin expression were detected by semi-quantitative RT-PCR and immunohistochemical SP methods. The effect of siRNA in suppressing the proliferation of PC-2 cells was detected by MTT assay, and its role in inducing PC-2 cell apoptosis evaluated by flow cytometry.

RESULTSThe sequence-specific siRNA effectively suppressed survivin expression at both mRNA and protein levels with inhibition rate of 81.25% at mRNA level and 74.24% at protein level. Survivin expression suppression significantly inhibited the proliferation of PC-2 cells, and at 24 and 48 h after cell seeding, the proliferation inhibition rate was 28.00% and 33.38% respectively; 24, 48 h after the transfection, apoptosis occurred in 8.46% and 7.53% of the cells, respectively.

CONCLUSIONSThe plasmid expression vector for the siRNA against survivin constructed in the study can effectively and specifically suppress survivin expression in PC-2 cells, and blocking survivin expression suppresses PC-2 cell proliferation and induces cell apoptosis. siRNA targeted against survivin has a potential value in gene therapy for pancreatic cancer.

Apoptosis ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Pancreatic Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVEBlocking the expression of survivin with RNA interference techniques, the effects of suppressing proliferation and inducing apoptosis of breast cancer MCF-7 cells were investigated.

METHODSA siRNA eukaryotic expression vector against survivin was constructed and transfected into breast cancer MCF-7 cells with lipofectamine 2000. The changes of survivin expression were detected by semi-quantitive RT-PCR and immunohistochemistry. The effect of suppressing proliferation of MCF-7 cell was detected by MTT assay. The effect of inducing MCF-7 cell apoptosis was detected by TUNEL assay.

RESULTSThe sequence-specific siRNA can efficiently block the expression of survivin both at mRNA and protein levels. The expression inhibition rate was 64.9% at mRNA level detected by semi-quantitive RT-PCR and 79.7% at protein level detected by immunohistochemistry. Blocking the expression of survivin can suppress proliferation of MCF-7 cells significantly. At 24 and 48 h after the cells were reseeded, the proliferation inhibition rates were 31.6% and 33.0%, respectively. At 24 h after transfection, apoptosis was induced in 12.9% of the cells as detected by TUNEL assay.

CONCLUSIONBlocking the expression of survivin with RNA interference technology can significantly suppress proliferation of MCF-7 cells and induce apoptosis to a certain degree. RNAi targeted to survivin has a potential value in gene therapy of breast cancer.

Apoptosis ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Genetic Therapy ; Genetic Vectors ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; physiology ; Neoplasm Proteins ; biosynthesis ; genetics ; physiology ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; metabolism ; pharmacology ; Transfection

OBJECTIVE: To explore the molecular basis for a Chinese family affected with neurofibromatosis type I. METHODS: Peripheral blood samples were collected from the proband and his parents. Potential mutations of NF1 gene were screened by PCR and Sanger sequencing. Pathogenicity of candidate mutations was analyzed using Polyphen-2 and Provean software. RESULTS: Two mutations of the NF1 gene, including c.702G>A (synonymous mutation) and c.1733T>G (missense mutation), were discovered in the proband. Neither mutation was found in his parents and 50 healthy controls. Bioinformatics analysis indicated that the c.1733T>G mutation (p.Leu578Arg) was probably damaging. The affected codon L578 is highly conserved across various species. CONCLUSION The c.1733T>C mutation of the NF1 gene probably underlies the neurofibromatosis type I in this family.
Asian Continental Ancestry Group ; Genes, Neurofibromatosis 1 ; Humans ; Mutation ; Neurofibromatosis 1 ; genetics ; Neurofibromin 1 ; genetics ; Pedigree

Objective To understand the influence of HIV infection on hepatitis C progress in patients co-infected with HIV and hepatitis C virus (HCV) and related immune mechanism. Methods Twenty eight patients co-infected with HIV/HCV and 12 patients with simplex HCV infection were enrolled. The liver function and hepatic fibrosis progress were evaluated by detecting peripheral blood and with Fibro-Scan. The viral load of HCV was detected by using real time quantitative PCR. And the percentage of Treg/CD4 +T lymphocyte cell was tested by using flow cytometry. Results The levels of ALT and ALP in HIV/HCV co-infection group were (76.16 ± 81.248)U/L,(24.507 1 ± 8.194)g/L respectively,higher than those of simplex HCV infection group [(27.475 0±13.985)U/L,(16.966 7±7.189)g/L],the differences were statistical significant. P value was 0.012 and 0.009 respectively. The liver fibrosis index in HIV/HCV co-infection group was 5.950 0-5.825 0 Kpa,higher than that in simplex HIV infection group(5.150 0-1.050 0 Kpa),and the difference was nearly statistical significant(P=0.077). The HCV viral load in HIV/HCV co-infection group was(6.476 8-5.343 4)lg copy/ml,higher than that in simplex HCV infection group[(1.699 0-2.681 5)lg copy/ml],and the rate of HCV clearance in HIV/HCV co-infection group was 32.14%, lower than that in simplex HCV infection group(75.00%). P value was 0.012 and 0.032 respectively. The percentage of Treg/CD4+T lymphocyte cell in HIV/HCV co-infection group was (7.460 0%-2.287 5%),higher than that in simplex HCV infection group (5.965 0%-2.105 0%),and the difference was significant (P=0.032). The percentage of Treg/CD4 + T lymphocyte cell was significantly related with HCV viral load(ρ=0.350,P=0.027),and HCV viral load was significantly related with the liver fibrosis index(ρ=0.487,P=0.001). Conclusion HIV infection could accelerate the progress of hepatitis C,and Treg cells were involved in this progress.