Analysis of ocular clinical features in 13 children with methylmalonic acidemia / 中华眼底病杂志

Objective:To observe the clinical features of eyes in children with methylmalonic acidemia (MMA).Methods:A retrospective clinical case study. From June 2019 to June 2022, 13 children with MMA visited on the Department of Ophthalmology of Henan Children's Hospital were included in the study. The anterior segment and fundus were examined under surface or general anesthesia. Best corrected visual acuity (BCVA) and refraction were performed in 9 cases; fluorescein fundus angiography (FFA) was performed in 3 cases; flash electroretinogram (FERG) was performed in 6 cases; flash visual evoked potential (FVEP) was detected in 6 cases; optical coherence tomography (OCT) was performed in 3 cases.Results:Among the 13 pediatric patients with methylmalonic acidemia, 6 cases were male and 7 cases were female. The average age at first visit was 45 months. All cases suffered from hyperhomocysteinemia; 9 cases were with epilepsy; 2 cases were with infantile spasms; 11 cases were with stunting, 13 cases were with repeated pulmonary infection during growth period; 4 cases were with hydrocephalus; 1 cases was with hypertension and renal insufficiency. Genetic dectection results of 8 cases were recorded, MMACHC:c.609G>A:p.W203* mutation site was found in all cases. One case was accompanied by corneal ulcer. There were 10 cases with nystagmus, 4 cases with macular degeneration, 3 cases with hyperopic refractive error and esotropia. Nine cases underwent BCVA examination, BCVA was light perception-0.6. In OCT, 2 cases of 3 cases showed retinal thinning and photoreceptor cell layer atrophy in the macular area. In FFA, 2 cases of 3 cases showed circular transparent fluorescence in the macular area. Five cases of 6 cases who with FVEP had different degrees of P100 peak time delay and decreased amplitude, and 4 cases of 6 cases with FERG had decrease of a and b wave in light and dark adaptation. Conclusions:The clinical phenotypes of eyes in children with MMA are various and the severity was different; most of them are accompanied by nystagmus, and the fundus lesions are common in the characteristic bovine eye like macular region. Those with macular disease have severe visual impairment.

Scutellarin inhibits caspase-11 activation and pyroptosis in macrophages via regulating PKA signaling

@#Inflammatory caspase-11 senses and is activated by intracellular lipopolysaccharide (LPS) leading to pyroptosis that has critical role in defensing against bacterial infection, whereas its excess activation under pathogenic circumstances may cause various inflammatory diseases. However, there are few known drugs that can control caspase-11 activation. We report here that scutellarin, a flavonoid from Erigeron breviscapus, acted as an inhibitor for caspase-11 activation in macrophages. Scutellarin dose-dependently inhibited intracellular LPS-induced release of caspase-11p26 (indicative of caspase-11 activation) and generation of N-terminal fragment of gasdermin D (GSDMD-NT), leading to reduced pyroptosis. It also suppressed the activation of non-canonical nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome as evidenced by reduced apoptosis-associated speck-like protein containing a CARD (ASC) speck formation and decreased interleukin-1 beta (IL-1β) and caspase-1p10 secretion, whereas the NLRP3-specific inhibitor MCC950 only inhibited IL-1β and caspase-1p10 release and ASC speck formation but not pyroptosis. Scutellarin also suppressed LPS-induced caspase-11 activation and pyroptosis in RAW 264.7 cells lacking ASC expression. Moreover, scutellarin treatment increased Ser/Thr phosphorylation of caspase-11 at protein kinase A (PKA)-specific sites, and its inhibitory action on caspase-11 activation was largely abrogated by PKA inhibitor H89 or by adenylyl cyclase inhibitor MDL12330A. Collectively, our data indicate that scutellarin inhibited caspase-11 activation and pyroptosis in macrophages at least partly via regulating the PKA signaling pathway.

Follow up of observation in patients with retinoblastoma under special circumstances / 中华眼底病杂志

Objective:To observe the retinoblastoma (RB) reexamination of children with new and recurrence retinoblastoma under special circumstances.Methods:From January 2, 2020 to March 15, 2020, 30 children with RB who had fundus examination in Henan Children's Hospital were enrolled in this study. Among them, 14 were male, 16 were female; 18 were monocular and 12 were binocular. The average age was 37.07±18.15 months. The mean age of initial diagnosis was 20.23±13.77 months. Two patients had a family history (6.67%). In 42 eyes, stage B, C, D and E were 7, 8, 20 and 7 eyes, respectively. Twenty-one eyes had finished the treatment course and 21 eyes were during treatment. All the children underwent RetCam fundus examination, orbital MRI, ocular B-ultrasound and so on. Whether the children had new tumor or recurrence at different treatment stages was observed.Results:Among 7 eyes in stage B, there was no recurrence or new tumor at the end of treatment or in the process of treatment. Among 8 eyes in stage C, there were 1 eye with new tumor and 1 eye with activity tumor at the end of treatment. Among 20 eyes in stage D, there were 1 eye with recurrence tumor at the end of treatment, 3 eyes with new tumor and 7 eyes with activity tumor at the end of treatment. Among 7 eyes in stage E, 5 eyes had eyeball enucleation and 2 eyes were receiving treatment; there were 1 eye with activity tumor at the end of treatment, 1 eye with recurrence tumor, 1 eye with activity tumor. Among 18 monocular eyes, there were 11 eyes in the treatment process, 2 eyes with new tumor, 1 eye with recurrence tumor and 3 eyes with activity tumor. Of the 24 binocular eyes, 10 were receiving treatment and there were 3 eyes with new tumor, 6 eyes with activity tumor. Twenty-one eyes had finished the treatment course, the average time required for follow-up was 3.71±0.31 months, and the average time delayed for follow-up was 6.43±1.66 weeks. There was a recurrence of tumor in 1 patient who had finished the whole treatment, the incidence was 4.76%. In the course of treatment, 21 eyes were required to have a follow-up time of 3 weeks, and the average delayed follow-up time was 6.00 ± 1.89 weeks. There were 5 eyes with new tumors, with a incidence of 21.74%. Nine eyes still had activity and needed to be treated in time.Conclusions:The higher the risk of tumor staging, the more relapses and new tumors. The patients who are being treated, the time of delayed follow-up, the higher the recurrence or new tumor than the children who have finished the treatment course and delayed the follow-up. The children who have relapsed or new tumor in the treatment course of binocular are higher than the children who have monocular.

Computer navigation-assisted surgical techniques in the surgical resection of pelvic chondrosarcomas / 中华骨科杂志

Objective:To explore the clinical effects of computer navigation-assisted surgery in the precise resection of pelvic chondrosarcoma.Methods:A retrospective analysis of 54 patients who had computer-assisted surgery from Dec 2007 to Dec 2018, including 27 males and 27 females, was conducted. The average age was 34.00±1.41 years (range 23-72 years). There were 47 cases with primary tumors and 7 with recurrence cases. The tumors in 15 cases located in the ilium (region I), 35 in the acetabulum (region II), 1 in the pubic (region III), and 3 in the sacroiliac joint (region IV). A total of 45 cases (83.3%) underwent needle biopsy, and 4 cases (7.4%) had incision biopsy. Among 5 cases who did not have biopsy, two of them was diagnosed of malignant change of multiple osteochondromas, two cases were diagnosed of recurrent pelvic chondrosarcoma and one with pelvic malignant tumor by imaging examinations. Pathological grade was presented as following, 36 cases in grade I, 15 in grade II, and 3 in grade III. All operations were performed on the bases of preoperative design with computer navigation-assisted surgical technology. A total of 49 cases (90.7%) had limb salvage operations and 5 cases had amputations. The surgical margins were confirmed by gross appearance and the maximum diameter profile of the tumor. Univariate analysis was performed to compare recurrence rate of different preoperative tumor status, gender, tumor stage, biopsy method, tumor location, operation method and surgical margins.Results:There were 39 cases underwent extensive resection, 13 cases with marginal resection and 2 cases with intracapsular resection. In 52 cases (96.3%), the surgery was performed according to the preoperative plan of surgical resection margin. However, two cases (3.7%) was not performed based on the preoperative plan. All patients were followed-up for 84.00±93.34 months (range 12-150 months). During the follow-up, a total of 45 cases (83.3%) survived and 9 cases died from lung metastasis. Eight cases (14.8%, 8/54) had local recurrence of whom 7 (14.3%) were limb salvage cases and 1 (20.0%, 1/5) had amputation. There was significantly different in local recurrence rate (χ 2=17.022, P=0.001). The risk of recurrence of marginal resection was 8.222 times than that of extensive resection [95% CI (1.297, 52.140)]. According to the Musculoskeletal Tumor Society (MSTS) limb function evaluation system score, postoperative limb function recovery rate was 90.00%±4.71% (range 60.00%-100%). There were 13 cases (24.1%) had postoperative complications, including 7 cases (13.0%) of infection, 2 cases (3.7%) of operative area and deep vein thrombosis of lower extremity, and 4 cases (7.4%) of skin necrosis and delayed healing. Among 49 limb salvage patients, two of them had secondary amputation due to tumor recurrence, five had hemipelvectomy due to neurovascular tumor invasion. The final limb salvage rate was 77.8% (42/54). Conclusion:Computer navigation-assisted precise pelvic tumor resection is technically feasible. It could decrease recurrence rate and promote limb function recovery by improving the reliability of oncology evaluation and the accuracy of tumor resection with superior safety.

Diarrhea pathogens in rotavirus-negative infants in Hangzhou / 中华检验医学杂志

Objective To investigate the distribution of diarrhea pathogens in infants without rotavirus-detection in Hangzhou. Methods 605 stool samples of children with rotavirus-negative diarrhea were collected from Hangzhou First People's Hospital of Zhejiang University, Children's Hospital of Zhejiang University and Hangzhou Children's Hospital from March 2017 to June 2018. The routine test results were analyzed retrospectively and Bristol score was used for the characteristics of stool samples. DNA and/or RNA were extracted from fecal samples with DNA and RNA extraction kit. The extracted DNA and RNA-reversed cDNA were used as templates. 7 common pathogens DNA and/or RNA were amplified by polymerase chain reaction (PCR). The amplified products were detected by agarose gel electrophoresis. The positive rates of pathogens were analyzed by chi-square test. Results Among 605 children, 375 were male (28±11) months and 230 were female (29±10) months. Bristol score of stool samples was mainly in type 6 (496, 82％), followed by type 7 (85, 14％) and type 5 (24, 4％). Among 605 results 97 cases were occult blood positive (positive rate 16％) and 170 cases were white blood cell positive (positive rate 28％).452 of 605 stool samples were positive for pathogen target genes. The positive rate was 74.7％. 319 cases detected single pathogen gene fragments. 127 cases detected two pathogen genes and 6 cases detected three pathogen gene fragments. The positive rate of Clostridium difficile toxin B (48.9％, 296/605)was the highest than the others, followed by Salmonella (20.0％, 121/605) and Norovirus (10.9％, 66/605). The positive rate of Clostridium difficile toxin A was 1.0％ (6/605). The positive rates of pathogens in male and female children were 86.7％(325 / 375) and 86.5％ (199 / 230) respectively, with (χ2 =0.002, P=0.959). Conclusions Salmonella and Norovirus were the main pathogens in children with diarrhea who were negative for rotavirus detection in Hangzhou. The high positive rate of Clostridium difficile toxin B may be related to the colonization of Clostridium difficile in the gastrointestinal tract of infants rather than the pathogen of diarrhea because of the low positive rate of Clostridium difficile toxin A. There was no gender difference in the detection rate of diarrhea pathogens.

Effects of laryngeal mask ventilation combined with continuous nerve block analgesia on postoperative lung infection in elderly patients undergoing the lower extremity orthopedics surgery / 中华老年医学杂志

Objective To investigate effects of laryngeal mask ventilation combined with continuous nerve block analgesia versus the endotracheal intubation under general anesthesia on postoperative lung infection in elderly patients undergoing orthopedic surgery,so as to provide a theoretical basis for a reasonable anesthesia method.Methods A total of 180 elderly patients undergoing lower extremity orthopedic surgery at our hospital from January 2016 to December 2016 were enrolled and randomly divided into two groups.The control group (n =90) were treated with endotracheal intubation under general anesthesia and intravenous analgesia after the operation,and the observation group (n =90)received the ultrasound-guided continuous nerve block analgesia in spontaneous respiration with laryngeal mask ventilation.The anesthetic dosage,awakening time,visual analogue scale(VAS)scores,and incidence of lung infections at 7 days after surgery were compared between the two groups.Pathogenic strains causing lung infections were isolated and identified.Results The anesthetic dosage was lower in the observation group than in the control group(P ＜ 0.05).The awaking time was shorter in the observation group than in the control group[(22.4±4.4) min vs.(34.1±8.5)min,P ＜0.05].The VAS scores under postoperative exercise were lower in the observation group than in the control group(P ＜0.05).The adverse reaction rate after surgery was lower in the observation group than in control group(3.3％ or 3/90 cases vs.41.1％ or 37/90 cases,P ＜0.05).The incidence of lung infections at 7 days after surgery was lower in the observation group than in the control group(3.3％ or 3/90 cases vs.11.1％ or 10/90 cases,P＜0.05).Ten pathogenic strains were isolated from control group,of which 9 strains were gram-negative bacteria,accounting for 90.0％.Three pathogenic strains were isolated from the observation group,of which 2 strains were gram-negative bacteria,accounting for 66.7％.Conclusions Laryngeal mask ventilation combined with continuous nerve block analgesia can reduce the anesthetic dosage,shorten the awaking time,provide a better analgesic effect and decrease the incidence of lung infections in elderly patients undergoing lower extremity orthopedics.The main pathogenic bacteria are gram-negative bacteria.

The effect of optical magnification during retinal nerve fiber layer thickness measurement in different degrees of myopia by using frequency domain OCT / 中华实验眼科杂志

Objective To investigate the effect of optical magnification on retinal nerve fiber layer (RNFL)thickness in different degrees of myopia by using spectral domain optical coherence tonmography (SD-OCT).Methods A retrospective case control study was designed.Eighty-five healthy people who underwent myopia correction and physical examination in the ophthalmology department of the Second Affiliated Hospital of Zhengzhou University from September to December in 2017 were selected,one eye was randomly selected from each subject.All the subjects were divided into 20 cases of emmetropic group,21 cases of low myopia group,20 cases of moderate myopia group,and 24 cases of high myopia group according to diopter.The visual acuity,best corrected visual acuity,slit lamp microscopy,fundoscopy,intraocular pressure and axial measurement,SD-OCT and visual field examination were performed on all the subjects.The difference of the mean RNFL and the thickness of the peripapillary quadrants among different groups,and relationship between RNFL and length of eye axis or diopter were analyzed before and after the correction of optical magnification.This study followed the Declaration of Helsinki.Results Before optical magnification correction,the average thickness of average,upper,lower and nasal quadrants RNFL were negatively correlated with the length of ocular axis (r =-0.595,-0.493,-0.639,-0.500;all at P =0.000),positively correlated with the diopter (r =0.005,0.565,0.600,0.464;all at P =0.000);the thickness of temporal quadrant RNFL was positively correlated with the length of ocular axis (r--0.683,P =0.000),negatively correlated with the diopter (r =-0.730,P =0.000).After optical magnification correction,the thickness of average,upper,lower and nasal quadrants RNFL had no correlation with the length of ocular axis and diopter (all at P＞0.05);the thickness of temporal quadrant RNFL was positively correlated with the length of ocular axis (r =0.840,P =0.000),negatively correlated with the diopter (r=-0.855,P =0.000).Before optical magnification correction,the thickness of average,upper,lower and nasal quadrants RNFL in emmetropic group were significantly higher than those of the other three groups (all at P＜0.05).The thickness of temporal quadrant RNFL was significantly lower than those in the other three groups (all at P＜0.05).The thickness of average,upper,lower and nasal quadrants RNFL in high myopia group were significantly lower than those in low myopia group and moderate myopia group (all at P ＜ 0.05),while the thickness of temporal quadrant RNFL was significantly higher than those in low myopia group and moderate myopia group (all at P＜0.05).There was no significant difference in RNFL thickness between low myopia group and moderate myopia group (all at P＞0.05).After optical magnification correction,the thickness of average,upper,lower and nasal quadrants RNFL showed no significant differences among the 4 groups (all at P＞0.05).The thickness of temporal quadrant RNFL was significantly different among the 4 groups (F =58.313,P =0.000).Conclusions When measuring RNFL thickness in myopic eyes by SD-OCT,the longer the axial length,the more obvious the optical magnification effect.The thickness of temporal quadrant RNFL increases in high myopia patients,so glaucoma should be vigilant when the thickness of temporal quadrant RNFL decreases.

Metformin enhances ATP-stimulated inflammasomeactivation in LPS-primed peritoneal macrophages / 中国药理学通报

Aim To explore the influence of metformin(a first-line drug for type 2 diabetes) on ATP-induced inflammasome activation and the release of interleukin-1β(IL-1β) by LPS-activated peritoneal macrophages, a commonly-used inflammatory cell model.Methods Peritoneal macrophages were elicited by intraperitoneal injection of 30 g·L-1 thioglycollate into C57BL/6 mice.Inflammasome was activated and cell pyroptosis was induced by LPS plus ATP treatment, and the pyroptotic cells were calculated after propidium iodide(PI) staining.The protein levels of IL-1β and caspase-1 expressed in the cells and released from them into the supernatant were evaluated by Western blot.Immunofluorescent microscopy was recruited to detect the subcellular distribution and fluorescent intensity of the purinergic P2X7 receptor(P2X7R).Results Metformin per se did not induce pyroptosis in LPS-activated peritoneal macrophages, but it significantly and dose-dependently increased cell pyroptosis induced by ATP treatment.At protein levels, maturated IL-1β(17 ku) could not be released from the cells upon single LPS or LPS plus metformin stimulation;but after ATP was added, maturated IL-1β was released into the supernatants of the cells.Moreover, metformin dose-dependently increased the protein levels of both maturated IL-1β and active caspase-1 released by the LPS-activated peritoneal macrophages upon ATP stimulation.Conclusion Metformin intensifies the activation of inflammasome and increases the release of active caspase-1 and maturated IL-1β upon ATP stimulation in the LPS-activated peritoneal macrophages, which should promote inflammatory responses.

Systematic analysis on legal system of workplace violence prevention and control in Chinese medical institutions / 中国职业医学

OBJECTIVE: To systematically analyze the related laws and regulations on workplace violence in Chinese medical institutions and provide recommendations for making policy. METHODS: The legal databases of CNKI,Wanfang,Lawyee and Pkulaw were retrieved by keywords including workplace violence,medical violence,medical dispute,doctor-patient dispute or medical order. The laws and regulations,department measures,local laws and government regulations were collected and systematically analyzed. RESULTS: In China,there are eight laws,two administrative regulations,two department rules and 15 provincial local laws and regulations stipulating that the healthcare workers enjoy the right to be free from workplace violence and constitute the legal basis for the prevention of medical violence in the workplace. During 2009-2017,the National Health and Family Planning Commission joints with multi-sector authorities have issued 12 notices related to the maintenance of medical order,from three aspects of human defense,physical defense and technical defense to vigorously emphasize the punishment of violent crime against personal safety of healthcare workers. However,compared to the international community,there is still a gap between China's legislation on the prevention and control of violence in medical workplaces,and there is no coherence in the definition of violence against the workplace. There are no specific legal and technical standards,and the occupational hazards caused by mental violence have not yet been included in the scope of legal adjustment and the national occupational disease classification directory. CONCLUSION: Specific legislation on prevention and treatment of medical institutions workplace violence should be carried out by using effective measures. The establishment of standard system,the related research and international exchange should be strengthened.

Cucurbitacin E induces autophagy in HeLa cells by inhibiting mTORC1 activity / 中国药理学通报

Aim To study the mechanism of cucurb-itacin E ( CuE )-induced autophagy in HeLa cells. Methods Improved MTT assay was adopted to meas-ure the effect of CuE on cell proliferation. Western blot was used to determine the phosphorylation levels of downstream signaling proteins of mTORC1 and the ex-pression of autophagy associated proteins. ResultsCuE inhibited the proliferation of HeLa cells in a dose-dependent manner, and the 24-h IC50 of CuE was 4. 01μmol· L-1 . CuE significantly inhibited the phospho-rylation of p70 S6 K in a time-and dose-dependent man-ner as evidenced by decreased phosphorylation levels of the mTORC1 substrate. Meanwhile, the expression of LC3-II, a marker for autophagosome formation, was elevated by CuE treatment, and was further increased in the presence of chloroquine. Furthermore, CuE re-duced the levels of p62/SQSTM1 . These results indi-cated that CuE induced autophagy in HeLa cells. The decreased levels of phosphorylated ULK1 S757 were posi-tively correlated with autophagy induction in HeLa cells. Conclusion CuE is likely to induce autophagy through inhibiting mTORC1 activity.

The clinical research of the dynamic external fixation stents treatment for contour type radius distal comminuted fracture / 重庆医学

Objective To observe the clinical effect of the dynamic external fixation stents treatment for contour type radius dis-tal comminuted fracture .Methods A total of 74 cases of patients with the contour type radius distal comminuted fracture were se-lected as the research object and were randomly divided into the observation group and the control group .37 cases of patients in the observation group underwent the dynamic external fixation stents treatment after closed reduction ;and 37 cases of patients in the control group underwent plaster external fixation treatment after closed reduction .Then the reset and functional effect of the two groups were compared after treatment .Results The patients of two groups were followed up for 1 year .The excellent and good rate of reset score in the observation group was 91 .89% ,which was higher than that of the control group(45 .95% ) ,the compara-tive differences was statistical significance(P=0 .000 0) .The excellent and good rate of function evaluation in the observation group was 86 .49% ,which was higher than that of the control group(40 .54% ) ,the comparative differences was statistical significance (P=0 .000 1) .Conclusion The dynamic external fixation stents treatment for contour type radius distal comminuted fracture can achieve satisfactory reset and functional recovery effect .

Establishment and application of DPO-based quadruple real-time PCR for detection of mycobacteria / 中华检验医学杂志

Objective To establish a rapid,accurate and specific method to detect the common mycobacteria based on multiplex real-time PCR.Methods The dual priming oligonucleotide ( DPO)primers and TaqMan probes labeled with FAM,ROX,HEX or JOE fluoresceins at 5' end and eclipse at 3' end respectively were designed to detect the 16S rRNA of mycobacteria.Both specificity and sensitivity were estimated on multiplex real-time PCR detecting genome DNA from 4 mycobacterial model species.Sixty eight early morning sputum specimens collected from hospitalized patients in the Red Cross Hospital of Hangzhou were detected by multiplex real-time PCR,bacterial culture and smear microscopy simultaneously.The positive rates were analyzed by chi-square.Results Mycobacteria including Mycobacterium tuberculosis and three common non-tuberculosis mycobacteria spp.were identified by multiplex real-time PCR accurately and specifically,with the limited load at 101 cfu/ml.In 68 sputum specimens,31 were positive (positive rate 45.6％ ) by this method,which was significant higher than that by smear microscopy ( positive rate 14.7％,x2 =15.4,P ＜0.05 ).The positive cases were identified as 28 Mycobacterium tuberculosis,1 Mycobacterium avium and 2 Mycobacterium intracellulare in agreement with the culture results.One case,which is detected by culture,but not by PCR,was identified as Mycobacterium chelonae by sequencing.Conclusion The multiplex real-time PCR characterizing as sensitive,specific and time-saving for Mycobacterium tuberculosis and common non-tuberculosis mycobacteria could be chosen as the rapid laboratory test of mycobacterial infection.

Computer navigation aided precision excision for sacrum tumor / 中华骨科杂志

Objective To assess the clinical significance of the application of computer assisted navigation technology in excision for sacrum tumor.Methods From December 2007 to June 2009,13 patients with sacrum tumor were treated with computer navigation assisted aggressive curettage.There were 5 males and 8 females.aged 21 to 69 years,with the mean age of 44 years,10 cases were over the level of sacrum3 and 3 cases under the sacrum3.Pathologic diagnosis was chordoma in 7 cases.giant cell tumor in 4 cases and neurofibroma in 2 cases.Five cases were recurrence.The preoperative data of CT and MRI were input into the computer navigation workstation.CT images determined the scope of the invasion of tumor in bone tissue.MRI determined the scope of the invasion of tumor in soft tissue.The CT and MRI image fusion identified the precise boundaries of the tumor in CT images and made markers for navigation guidance in the operative in 10 cases.The Iso-c scan had been made for another 3 cases and fused the Iso-c images with preoperation CT images for improving the images quality.According to preoperative marker in CT images,the aggressive curettage were completed with the real time computer navigation for 7 cases,marginal resection in 4 cases and wide resection in 2.The precise surgical removal of the lesions boundary were verified by computer navigation according to the preoperative planning.Results Thirteen patients were followed up for 7 to 37 months,an average of 18 months,no recurrence cases with marginal resection and wide resection.Two cases with aggressive curettage had recurrence.Conclusion Computer navigation technology helps to precise excision the sacrum tumors,to reduce the recurrence rate.

Valproic acid exerts differential effects on cytokine synthesis in human peripheral lymphocytes / 中国病理生理杂志

AIM: Valproic acid (VPA) is a histone deacetylase inhibitor and is believed to have anti-tumor activity. The present study aims to investigate the effect of VPA on the, apoptosis and cytokine synthesis of human peripheral lymphocytes. METHODS: The activation and cytokine synthesis in lymphocytes in whole blood stimulated with phorbol dibutyrate (PDB) and ionomycin were evaluated with flow cytometry after fluorescent staining. The mitochondrial membrane potential was examined using 3, 3-dihexyloxacarbocyanine iodide [DiOC6(3)]staining. RESULTS: VPA at low doses (1 and 5 mmol/L) promoted CD69 expression in activated lymphocytes, whereas it turned to inhibit the expression of CD69 at a high dose (25 mmol/L). Meanwhile, VPA at low doses increased the mitochondrial membrane potential, while a high dose of VPA decreased it in activated lymphocytes. Furthermore, interleukin-2 (IL-2) synthesis was enhanced by low doses of VPA but inhibited by a high dose. However, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) synthesis were dose-dependently enhanced by VPA as compared with those of PDB plus ionomycin-treated cells. CONCLUSION: VPA exerts biphasic effect on the further activation and apoptosis of human peripheral lymphocytes stimulated with mitogens and exhibits differential activity on the synthesis of several important cytokines in human lymphocytes.

Fluorescence PCR for detecting ureA gene of Helicobacter pylori in feces / 中华临床感染病杂志

Objective To evaluate the application of fluorescence PCR in detecting ureA gene of Helicobacter pylori(HP)in feces.Methods Fluorescence PCR was used to detect ureA gene of HP in feces from 50 patients,including 23 confirmed by gastric biopsy urease test and histological staining.Bacterium culture and serum antibody detection were also performed, and chi-square test was used to compare the sensitivity,specificity,positive predictive value and negative predictive value among three methods.Results The sensitivity,specificity,positive predictive value and negative predictive value of fluorescence PCR were 1.00,0.96,96.O%and 100.0%,while those for HP culture were 0.78,1.00,100.0%and 84.0%,and thee for serum antibody detection Was 0.96,0.74,76.O%and 95.0%.There were significant differences in sensitivity and negative predictive value between PCR and bacterium culture (X2=5.60 and 4.44,P<0.05),and significant differences in specificity and positive predictive value between PCR and serum antibody detection(X2=5.28 and 4.08,P<0.05).Conclusion ureA gene detection in feces by fluorescence PCR is of value for the diagnosis of HP infection.

Role of Helicobacter pylori cheA gene in chemotaxis in vitro and colonizationin vivo / 中华微生物学和免疫学杂志

Objective To determine the effect of cheA gene of Helicobacter pylori in the bacterial chemotaxis in vitro and colonization in vivo. Methods The entire cheA and cheY genes were amplified and cloned from genomic DNA of H. pylori NCTC11637 strain. Subsequently, the prokaryotic expression systems of cheA and cheY genes were generated and the target recombinant proteins rCheA and rCheY were extracted by Ni-NTA affinity chromatography. Rabbits were immunized with either rCheA or rCheY for obtaining antisera, and rCheA-IgG and rCheY-IgG in the antisera were prepared using ammonium sulfate precipitation plus DEAE-52 column chromatography. A suicide plasmid of cheA gene was constructed and then a cheA gene knock-out mutant ( cheA - ) was generated based on homologous recombinant exchange using the suicide plasmid. The cheA- mutant was identified using PCR and sequencing. The phosphorylation levels of CheA and CheY molecules of cheA - and wild-type strain were determined by using rCheA-IgG and rCheY-IgG anchoring the target proteins and protein phosphorylation detection kit. The differences of chemotaxis in vitro and colonization in vivo between cheA- mutant and wild-type strain were compared using chemotactic model and BALB/c infection model of H. pylori. Results The cheA gene knock-out in genome of cheA- mutant was confirmed by the results of PCR and sequencing. After treated with 0. 001-0. 1 mol/L HCI for 10 min, the phosphorylation levels of CheA and CheY molecules of wild-type strain were rapidly descended from ( 59.6 ±11.5) μmol and (55.5 ± 10.2) μmol to ( 10.8 ± 2.6) and (5. 5 ± 1.2) μmol (P ＜ 0.05 ), while the phosphorylation of CheY molecule of cheA - mutant was no markedly changed with a persistent lower level ( P ＞0.05). The diameters [(10-20) ± (2-3) mm] of chemotactic aggregative rings of cheA- mutant were significantly less than those [(16-24) ± (2-3)mm] of wild-type strain (P ＜0.05). The positive isolation rate (90%) of H. pylori in gastric biopsy specimens of mice that infected with wild-type strain was remarkably higher than that (40%) of mice that infected with cheA- mutant (P ＜0.05). The result of fluorescence quantitative was also showed that the numbers (6.3 × 103 ±2.1 × 103 copies/mg) of H. pylori in gastric biopsy specimens of wild-type strain infected mice were significantly larger than those (8.3 × 101 ±3. 1 × 101 copies/mg) in gastric biopsy specimens ofcheA- mutant infected mice (P＜0.05). Conclusion The cheA gene of H. pylori has an important role in the bacterial chemotaxis in vitro and colonization in vivo.

Experssion of human FGFR2IIIc and its S252W mutant in MDA-MB-231 breast cancer cells with adenovirus vectors / 生物工程学报

To study the functions of human Fibroblast growth factor receptor 2IIIc (FGFR2IIIc) gene in cancer cells, breast cancer cells MDA-MB-231 were infected by recombinant adenoviruses containing FGFR2IIIc and its S252W mutant, respectively. FGFR2IIIc gene was amplified from an existing plasmid and its S252W mutant was obtained by overlapping extension PCR. These two genes were separately cloned into the adenoviral shuttle plasmid pAdTrack-CMV, confivmed by DNA sequencing linearized, and co-transformed into Escherichia coli BJ-5183 with the adenoviral vector pAdEasy-1. The resulting recombinant expression vectors Ad-FGFR2IIIc and Ad-FGFR2IIIcS252W were linearized and transfected into HEK293A cells to get adenoviral particles. GFP was used to verify the gene expression. The recombinant adenoviral particles were harvested, titrated, and then infected MDA-MB-231 cells. The expression of FGFR2IIIc and its S252W mutant were examined by RT-PCR and Western blotting, and the effect of these recombinant adenoviruses on MDA-MB-231 cell proliferation was analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry. The results showed the recombinant adenoviral particles could infect MDA-MB-231 cells and express the target proteins. MTT showed that both FGFR2IIIc and its S252W mutant inhibited MDA-MB-231 cell proliferation, but the mutant was more effective. Flow cytometry showed that both FGFR2IIIc and its S252W mutant arrested MDA-MB-231 cell cycle at G0/G1 phase, resulting in low cell proliferation.

Prokaryotic expression of Helicobacter pylori cheA and cheY genes and correlation among the expressed products and bacterial chemotactic behavior / 中华微生物学和免疫学杂志

Objective To clone the cheA and cheY genes of Helicobacter pylori for construction of their prokaryotic expression systems, and to establish chemotactic model in vitro of H. pylori for determing chemotaxis-inducing substances and to understand the effects of specific antibody and closantel on inhibiting chemotactic behavior of the microbe. Methods The segments of entire cheA and cheY genes were amplified by PGR and then sequenced after T-A cloning. Prokaryotic expression systems of the genes were subsequent-ly constructed. SDS-PAGE plus Bio-Rad Gel Image Analyzer were used to examine the expression of target recombinant proteins rCheA and rCheY, and Ni-NTA affinity chromatography was performed to extract rCheA and rCheY. Rabbits were immunized with rCheA and rCheY to obtain antisera and IgG in each of the anti-sera was extracted by saturated ammonium sulfate precipitation and DEAE-32 ion exchange chromatography. Immunodiffusion assay was performed to measure the titers of antisera and their IgGs. Chemotactic model in vitro of H. pylori based on hard-agar plus method was established to determine the chemotaxis-inducing effects of eleven candidate substances. Simultaneously, the effects of rCheA-lgG and closantel sodium on blocking the bacterial chemotactic behavior were also observed. Results The segments with expected sizes of cheA and cheY genes were obtained by PCR, and their nucleotide and putative amino acid sequences were 100% idenities to the reports. The constructed prokaryotic systems could efficiently express rCheA and rCheY. The two rabbit antisera and IgG aginst rCheA and rCheY had 1 : 4 and 1 : 2 immunodiffusion titers, respectively. Hydrochloric acid, sulfuric acid and acetic acid were able to induce chemotactic movement of H. pylori. Both rCheA-IgG and closantel sodium with certain concentrations could weaken the chemotactic ability of H. pylori(P<0.05). Conclusion The prokaryotic expression systems of H. pylori cheA and cheY genes were successfully generated in this study. Hydrogen ion (H~+) is the inducer for chemotaxis of H. py-lori. rCheA-IgG, as well as closantel sodium can inhibit H~+-induced chemotaxis of H. pylori.

Preparation of H-2Db tetramer and its application in enumerating the CD8+ T cells specific for lymphocytic choriomeningitis virus / 生物工程学报

Major histocompatibility complex (MHC) tetramer technology offers a powerful means to study specific T cell populations of interest. To investigate the immune response of H-2Db-restricted CD8+ T cells in immunotherapy, we prepared the H-2Db tetramer and verified its effectiveness in enumerating the CD8+ T cells specific for the lymphocytic choriomeningitis virus (LCMV). First, the cDNA encoding H-2Db heavy chain was cloned by RT-PCR from the spleen of a C57BL/6 mouse. The expression vector for H-2Db-BSP, i.e. the ectodomain of H-2Db fused to a BirA substrate peptide (BSP), was constructed and overexpressed in E. coli BL21(DE3). Then, the denatured H-2Db-BSP was refolded in the presence of human beta2-microglobulin as well as the GP33-41 peptide (KAVYNFATC, KAV) of LCMV. The biotinylated H-2Db/KAV molecules were purified, then bound to streptavidin-PE and tetramerized. Finally, the prepared H-2Db KAV tetramer reagent was verified by detecting the CD8+ T cells specific for HCMV in KAV peptide vaccinated C57BL/6 mouse, with a mouse receiving subcutaneous injection of only adjuvant as negative control. The results showed that the tetramer positive rates were 0.27%, 0.11%, and 0.24% within the CD8+ T cell populations in the peripheral blood, draining lymph nodes, and spleen of vaccinated mouse, respectively. There was only very low background staining (< or = 0.01%) of those samples from the control mouse. Beside, the best results were achieved in the staining of the peripheral blood sample. In conclusion, the established procedure of preparing H-2Db tetramer will facilitate the study of the immune responses of antigen-specific CD8+ T cells in the experimental immunotherapy on the mice with H-2Db allele background.

Application and evaluation of PBL teaching model in experiment teaching of medical university / 中华医学教育探索杂志

In order to meet the requirement of the innovation of higher education and fulfill the ideas of quality-oriented education, Biochemistry department of Hebei Medical University adopts PBL teaching approach.It is shown that PBL teaching approach can stimulate the students' interest and motivation in study,develop their self-study ability,activate their mind and improve the teaching quality.