Objective:The aims of this study are to detect the apoptosis of cancer cells after a single dose of radiotherapy with 99mTc-HYNIC-annexin V and to investigate the correlation among early apoptosis, radio-therapeutic dose, and radio-sensitivity. Methods:Ten days after respective inoculations of EL4 lymphoma and S180 sarcoma in their right upper limbs, the mice were randomly divided into imaging group (Group One) and observation group (Group Two). In Group One, 99mTc-HYNIC-annexin V was injected via the caudal vein after different doses of irradiation. Approximately 2 h later, clinical imaging was conducted by single-photon emission-computed tomography. The mice were sacrificed to evaluate the bio-distribution of 99mTc in each specimen. Cell count during apoptosis was conducted through the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method. Observation was conducted in Group Two for 2 weeks after the irradiation. Results:Tumor uptake of 99mTc-HYNIC-annexin V significantly correlated with the number of TUNEL-positive cells, which concordantly increased with the increase of dosage (r=0.892, P=0.000). With the same dose (0 or 8 Gy), the values of%ID/g, T/B, T/M, and TUNEL-positive cell number of EL4 lymphoma were significantly higher compared with those of S180 sarcoma. EL4 lymphoma was entirely minimized after irradiation at 8 Gy. Conclusion: 99mTc-HYNIC-annexin V can detect early apoptosis in vivo in tumors receiving radiation. The irradiation-induced apoptosis in vivo determined with 99mTc-HYNIC-annexin V positively correlates with the curative effect of tumors. The detection of tumor cell apoptosis via 99mTc-HYNIC-annexin V helps estimate radio-sensitivity, and can become a predictive index for radiotherapy.

Objective To explore the inhibitory effects of endogenous hydrogen sulfide, a novel and important gas-eous transmitter generated in mammalian tissues mainly by cystathionine β-synthase (CBS) or cystathionineγ-lyase (CSE) on the apoptosis of the rat hepatic BRL cell line in physiological condition. Methods BRL cells were cultured, and divid-ed randomly into several groups in different phases of the experiment, including negative-siRNA (control) group, CBS siRNA (CBS 1~3) group and CSE siRNA (CSE 1~3) group, which were used to select the most efficient sequences of siRNAs at 48 or 24-hour transfection. Solution group and (CBS+CSE) siRNA group were added to detect the variation of apoptosis. The BRL cell line was observed and evaluated at 0, 4, 8, 12, and 24 hrs after siRNA transfection. When the mechanisms of the apoptosis were detected, CBS/CSE siRNAs were transfected individually or jointly into BRL cells, and compared with nega-tive-siRNA group to examine the variation. The genic and protein expression of CBS/CSE were detected by RT-PCR and Western blot assay. After transfection of CBS/CSE siRNA, the apoptosis of BRL cells was detected by Hoechst stain and flow cytometry (FCM). The mitochondrial membrane potential (MMP) changes were observed by fluorescent staining. Western blot assay was used to examine the protein expression of intracytoplasm cytochrome C (Cyt C) and cleaved-caspase 3. Re-sults CBS and CSE were observed in BRL cells. After transfection of CBS/CSE siRNA, endogenous H2S generation de-creased and the apoptosis of BRL cells increased. Accordingly, the expression of intracytoplasm-Cyt C and cleaved-caspase 3 increased. Conclusion The inhibition of endogenous H2S synthesis induced the apoptosis of BRL cells under physiologi-cal condition, which may be involved in mitochondrial pathway of apoptosis.

AIM To investigate whether dipfluzine (Dip) possesses antiarrhythmic effect on experimental arrhythmias and effect on cytosolic calcium in ventricular myocytes of guinea-pig. METHODS Experimental arrhythmias were induced by strophanthin G infusion through jugular vein in guinea-pigs and by myocardial ischemia-reperfusion (I-R) in rats respectively. Cytosolic calcium concentration ([Ca2+]i) of isolated guinea-pig ventricular myocytes was examined with laser confocal scanning microscope. RESULTSIn guinea-pigs pretreatment with Dip 20 mg·kg-1 increased the dosages of strophanthin G required to induce ventricular premature contraction (VP), ventricular tachycardia (VT), ventricular fibrillation (VF) and cardiac arrest (CA), pretreatment with Dip 10 mg·kg-1 increased the dosages of strophanthin G required to induce VP. In the I-R-induced arrhythmic model of rats, Dip 20 mg·kg-1 decreased the number of rats exhibiting VT, VF and CA, and the number of rats exhibiting VF and CA was decreased by Dip 10 mg·kg-1. Both Dip and verapamil (Ver) decreased [Ca2+]i of the ventricular myocytes in normal Tyrode′s solution. The Ca2+ overload evoked by high extracellular Ca2+ levels was inhibited by Dip and Ver, and the prophylactic effect of Dip was less than that of Ver, while the curative effect of Dip was more obvious than that of Ver. CONCLUSION Dip has antiarrhythmic effect, which is likely related to the modulation on the intracellular calcium homeostasis.

OBJECTIVE: Oral zhongfeng an liquid is a new dose form of traditional Chinese medicine for treating cerebrovascular diseases. Its main components are astragalus and hirudo, the former is of obvious effects of replenishing qi and activating blood, and the latter is of stronger effects of antiplatelet, antithrombosis and arteriospasm-reducing, as well as improving tissue anoxia.OBJECTIVE: To investigate the effects of oral zhongfeng an liquid on arterial thrombosis in rats, and blood coagulation and tolerance in mice.DESIGN: A completely randomized and controlled trial.SETTING: Pharmacological Division of Basic Medical College, Hebei Medical University.MATERIALS: The experiment was completed from September 2001 to April 2002 at the Pharmacological Division of Basic Medical College,Hebei Medical University. The experiment of effect of the drug on thrombosis in rats: In the first study, totally 40 Wistar rats were at random divided into five groups: zhongfeng an liquid 3.0, 6.0, 12.0 g/kg, aspirin 0.3 g/kg and control group, with 8 in each group. In the second study, totally 50 Wistar rats were also at random divided into five groups: zhongfeng an liquid 3.0 and 6.0 g/kg, naoxue kang 3.0 and 6.0 g/kg and control group,with 10 in each group. Clotting time study in mice: Totally 50 mice were randomly divided as zhongfeng an liquid 6.0, 12.0, 24.0 g/kg, aspirin 0.3 g/kg and control group, with 10 in each group. Measurement of swimming exhaustion time of mice: Totally 90 mice were randomly divided as zhongfeng an liquid 6.0 and 24.0 g/kg, naoxue kang 6.0 and 24.0 g/kg, benzedrine 0.2 g/kg and control group, with 15 in each group.METHODS: In the experiment of effect of the drug on thrombosis: For the first study, 24 hours and 1 hour before operation the rats in all groups were respectively by gavage given oral zhongfeng an liquid (3, 6, 12 g/kg), aspirin (0.3 g/kg), and water. For the second study, 24 hours and 1 hour before operation the rats were respectively by gavage given oral zhongfeng an liquid (3, 6 g/kg), naoxue kang (3, 6 g/kg) and water. After administration,ketamine 50 mg/kg was peritoneally given for anesthesia, silk ligature thrombosis was used, then the wet thrombus was weighed for comparison of difference in thrombosis among the groups. Measurement of clotting time of mice: The mice were respectively by gavage given oral zhongfeng an liquid (24.0, 12.0, 6.0 g/kg), aspirin (0.3 g/kg) and water, one hour after administration the clotting time of mice was detected with the slide method. Measurement of swimming exhaustion time of mice: The mice were respectively by gavage given oral zhongfeng an liquid (6.0, 24.0 g/kg), naoxue kang (6.0, 24.0 g/kg), Benzedrine (0.2 g/kg) and water, once a day for 5 days.On the fifth day 1 hour after administration, the swimming exhaustion time of mice was measured, the mean value of swimming exhaustion time of mice in each group was calculated.haustion time of mice in each group.RESULTS: All 90 rats and 140 mice involved entered into the result thrombus in rats of the oral zhongfeng an liquid 3.0, 6.0 and 12.0 g/kg groups were obviously lower than those in the naoxue kang groups of the same dose [(24±4), (21±4), (16±6), (39±7) mg, (t=5.88-7.90, P ＜ 0.01)]; in the second study, the wet quality of rats in the oral zhongfeng an liquid 6.0 g/kg group was obviously lower than that in the same dose naoxue kang group [(23.6±2.6), (30.0±4.1), (t=4.18, P ＜ 0.01)], the wet quality of mice in the oral zhongfeng an liquid 3.0 g/kg group was obviously lower than that in the same dose naoxue kang group [(30.6±2.1), (33.1±1.6) mg, (t=2.96,zhongfeng an liquid 24.0 and 12.0 g/kg groups were obviously higher than that in the control group [(222±66), (190±52), (116±26) s, (t=4.02, 4.72, P the oral zhongfeng an liquid 24.0 and 6.0 g/kg groups were obviously higher than that in the control group [(2 512±1 244), (899±403), (502±100) s,(t=3.70-6.24, P ＜ 0.01)].CONCLUSION: Oral zhongfeng an liquid was of obvious inhibition to arterial thrombosis of rats and venous thrombosis of mice, and could enhance the tolerance of mice with a role of antifatigue.

Objective:To explore the investigation level of individual monitoring for medical radiation workers.Methods:Monitoring and analysis of individual doses to the medical radiation workers in Guangdong province were performed, from 2016 to 2019, by the Individual Dose Monitoring Department of Guangdong Provincial Hospital for Occupational Disease Prevention and Control.Results:The numbers of monitored workers were diagnostic radiology 53 674, dental radiology 2 563, nuclear medicine 5 001, radiotherapy 16 687, interventional radiology 22 272 and others 2 087 from 2016 to 2019, of which the number of individuals with doses in excess of investigation level 1.25 mSv, were 76, 6, 18, 28, 133 and 2 respectively. The non-real doses made up 67.1 %, 100 %, 55.6 %, 82.1 %, 76.7 % and 100 % of their respective totals. Their 99th percentile doses P99 were 0.37, 0.39, 0.67, 0.35, 0.54 and 0.30 mSv, and the average periodic dose equivalent were 0.07, 0.06, 0.11, 0.06, 0.07 and 0.05 mSv respectively, indicating a statistically significant difference existing in the average annual effective dose between groups ( Z=-26.139--2.681, P<0.001). Conclusions:Due to non-reality of doses in excess of investigation levels and high labor cost, it is suggested to currently use 0.40 mSv per 3 months as investigation level for diagnostic radiology, dental radiology, radiotherapy and others, and 0.70 mSv per 3 months as for nuclear medicine and interventional radiology.

We transformed the fip-fve gene into Pichia pastoris GS115 for inducible and constitutive expression to obtain feasible bioactvie recombinant Fip-fve. The fip-fve gene was cloned from Flammulina velutipes fruting body by PCR and ligated to pPIC9 to construct inducible expression vector pPIC9-FIP-fve, and promotor pgap was used to replace the paox1 to construct constitutive expression vector pPIC9-PGAP-FIP-fve. These two vectors were used to transform P. pastoris by PEG method. The fip-fve was expressed after histamine-absence screening and yeast colony PCR. The inducible expression level reached 158.2 mg/L at the fourth day and the constitutive expression level was 46.3 mg/L and 29.5 mg/L using glucose and glycerol, respectively. The SDS-PAGE and Western blotting both proved the correctness of rFip-fve, and the hemagglutination test indicats the rFip-fve's bioactivity.
Electrophoresis, Polyacrylamide Gel ; Flammulina ; chemistry ; Fungal Proteins ; biosynthesis ; Genetic Vectors ; Pichia ; metabolism ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Recombinant Proteins ; biosynthesis

OBJECTIVE: To study the expression of CD44 and nm23-H1 gene proteins and their clinical significance in laryngeal carcinoma. METHOD: The expression of CD44 and nm23-H1 proteins were detected by immunohistochemistry method in 40 cases with laryngeal carcinoma, 20 adjacent carcinoma tissues and 12 cases normal laryngeal mucosa tissues. RESULT: The expression of CD44 and nm23-H1 proteins in laryngeal carcinoma were much higher than that in normal laryngeal mucosa. The expression of CD44 protein in laryngeal carcinoma with metastatic lymph node was higher than that in laryngeal carcinoma without metastatic lymph node, but nm23-H1 protein lower. The expression of CD44 protein was positively correlated with the metastasis, clinical staging and pathological classification but not correlated with T classification of laryngeal carcinoma. The expression of nm23-H1 protein was negative correlation with the metastasis and clinical staging of laryngeal carcinoma. CONCLUSION CD44 and nm23-H1 gene proteins play an important coordinated regulation role in the carcinogenesis, development and metastasis of laryngeal carcinoma and will probably become the key biological marks in the judging and evaluating prognosis of laryngeal carcinoma.
Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Hyaluronan Receptors ; metabolism ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; metabolism ; Neoplasm Staging

Objective The purpose of this study was to explore common complications and their clinicopathological features in pediatric liver transplantation.Methods Clinical and pathological data of 240 liver biopsies from 168 children that conducted liver puncture from January 2015 to May 2018 in Tianjin First Central Hospital was retrospectively analyzed.We comprehensively analyzed incidence rate and pathological features of various complications,and correlations between acute rejection and C4d staining result or Banff score.Results A total of 86.67％ (208/240) liver biopsies could be definitely diagnosed with incidence rate of main complications in descending order as follows:T cell mediated rejection (TCMR) 60.57％ (126/208),drug-induced liver injury (DILI) 17.31％ (36/208),biliary complication 8.17％ (17/208),vascular complication 3.37％ (7/208),ischemia/reperfusion injury (IRI) 2.88％ (6/208),antibody mediated acute rejection (AMR) 1.92％ (4/208),HBV infection 1.92％ (4/208),non-alcoholic fatty liver disease (NAFLD) 1.44％ (3/208),chronic rejection (CR) 0.96 ％ (2/208) and HCV infection 0.48 ％ (1/208).TCMR and AMR in acute rejection (AR) accounted for 96.92％ (126/130) and 3.08％ (4/160),and into(portal-based,PB)type TCMR accounted for 96.03％(121/126) with the detectable rate of BP type subtype TCMR of 26.45％(32/121)within 30 d.There were 65.87％ (83/126)、25.40％ (32/126) 和4.76％ (6/126) of BP TCMR samples with "Banff ACR RAI" score within 3-5,6-7 and 8-9,and RAI score was negatively correlated with postoperative time (r =0.127,P =0.084).The incidence rate of central perivenulitis (CP) and portal eosinophils infiltration (PEI) in BP TCMR was 63.63％ (77/121) 和43.80％ (53/ 121),respectively,additionally,the PEI level was positively correlate with RAI score (P＜0.05).CP TCMR and AMR occurred within 30d-365 d and 8 d-180 d,respectively postoperative,while,the two CR occurred at 1095 d and 1335 d postoperative,and significant correlation was strikingly observed between rejection subtype and postoperative time (Z =9.231,P =0.026).C4d positive rate was 10％ (24/240),which was associated with Banff score and postoperative time,besides,C4d score was also correlated with rejection subtype and RAI score.The occurrence of DILI was mainly at time of ＜90 d or ＞180 d postoperative,and the detectable rate of biliary complication within 180 d postoperative was 82.35％ (14/17),IRI Appear in ＜30d.Hepatic artery complication account for nearly 57.14％ (4/7),occurrence time is ≤90 d.Occurrence of HBV infection,CMV infection and NAFLD were mainly at ＞365 d,＜90 d and ＜365 d,respectively.Conclusion There were lots of differences in clinical and pathological features among multi pediatric liver transplantation complications.Liver puncture plays an important role in rejection subtype classification and grading,as well as in non-rejection complications identification.

Objective:To evaluate the difference of dosimetry between three-dimensional and two-dimensional plans based on CT images of occult perforation in brachytherapy of cervical cancer, aiming to provide clinical reference.Methods:A total of 817 patients with cervical cancer received simple intrauterine (intrauterine tandem plus vaginal colpostats) three-dimensional brachytherapy in Chongqing University Cancer Hospital from January 2019 to December 2020 were retrospectively reviewed. Among them, 16 patients had occul uterine perforation. Based on Oncentra Brachy Therapy plan system, the single prescription dose was 6Gy. Three-dimensional (3D group) and two-dimensional (2D group) plans were designed on the perforated CT images The target volume, conformal index (CI), conformal index coformity index (COIN) and organs-at-risk (OAR) D 2cm 3 parameters were used to assess the plans between two groups. Results:The incidence of pccult uterine perforation was 1.96%(16/817) during brachytherapy for cervical cancer. The volume of prescription dose curve in the 3D group was (40.74±14.98) cm 3, significantly smaller compared with (91.46±19.71) cm 3 in the 2D group ( P<0.05), whereas the volume of the high-risk clinical target area wrapped by prescription dose curve did not significantly differ between two groups ( P>0.05). The CI and COIN in the 3D group were 0.79±0.10 and 0.72±0.96, significantly higher compared with 0.38±0.09 and 0.37±0.18 in the 2D group (both P<0.05). The D 2cm 3 of bladder, rectum, sigmoid colon, small intestine in the 3D group were (306.06±77.57) cGy, (252.27±72.60) cGy, (127.25±62.84) cGy and (228.79±94.90) cGy, significantly lower than (548.03±164.21) cGy, (411.16±118.74) cGy, (227.45±94.48) cGy and (450.95±157.96) cGy in the 2D group (all P<0.05). Conclusions:Application of image guidance in brachytherapy of cervical cancer is helpful to detect occult uterine perforation. When occult uterine perforation occurs, the use of three-dimensional plan can basically meet the clinical needs, which is significantly better than the two-dimensional plan.

Background: Cervical cancer is still present a major public health problem, especially in developing countries. In International Federation of Gynaecology and Obstetrics 2018, allowing assessment of retroperitoneal lymph nodes by imaging and/or pathological findings and, if deemed metastatic, the case is designated as stage IIIC (with r and p notations). Patients with lymph node metastases have lower overall survival (OS), progression free survival (PFS), and survival after recurrence, especially those who have unresectable macroscopical positive lymph nodes. Retrospective analysis suggests that there may be a benefit to debulking macroscopic nodes that would be otherwise difficult to sterilize with standard doses of radiation therapy. However, there are no prospective study reporting that resecting macroscopic nodes before concurrent chemoradiation therapy (CCRT) would improve PFS or OS of cervical cancer and no guidelines for surgical resection of bulky lymph nodes. The CQGOG0103 study is a prospective, multicenter and randomized controlled trial (RCT) evaluating lymph node dissection on stage IIICr of cervical cancer. Methods Eligible patients are histologically confirmed cervical squamous cell carcinoma, adenocarcinoma, adeno-squamous cell carcinoma. Stage IIICr (confirmed by computed tomography [CT]/magnetic resonance imaging/positron emission tomography/CT) and the short diameter of image-positive lymph node ≥15 mm. 452 patients will be equally randomized to receive either CCRT (pelvic external-beam radiotherapy [EBRT]/extended-field EBRT + cisplatin [40 mg/m2] or carboplatin [the area under curve=2] every week for 5 cycles + brachytherapy) or open/minimally invasive pelvic and para-aortic lymph node dissection followed by CCRT. Randomization is stratified by status of para-aortic lymph node. The primary endpoint is PFS. Secondary endpoints are OS and surgical complications. A total of 452 patients will be enrolled from multiple hospitals in China within 4 years and followed up for 5 years.