Objective To establish a method for determining the content of ceram ide in skin epidermis. Methods Epidermal lipids of mice were extracted by routin e method,separated by thin-layer chromatography (TLC),and shot by BIO-RAD Ge lDoc system. Epidermal ceramide content was determined by Software of Quantity O ne. Results A good linearity was obtained in the range of 0.416～2.08 ?g (r=0.9 95 3). The precision of the same plate was good,RSD=1.09 %;and the precision of different plates was also good,RSD= 2.10 %. The stability of control soluti on and sample solution was good within 2 h. The reproducibility was good,RSD= 3 .03 %,and the mean recovery rate was 99.58 %and RSD=1.46 %. The RSD of simpl e determination was 3.10 %. Conclusion This method can be used to determine the content of ceramide in skin epidermis.

Objective To investigate the relationship between the level of 8-iso-prostaglandin (8-iso-PG)F2αand the neural functional deficit in patients with acute cerebral infarction (ACI). Methods Sixty-seven ACI patients were se-lected in Neurological Department of Ganyu People’s Hospital. According to the age, these subjects were divided into two groups:the old group (≥60 years, n=37) and middle-young group (<60 years, n=30). Thirty healthy subjects were selected as controls (≥60 years). The age, gender and anamnesis were matched in two groups of elderly people. The ELISA was used to detect the plasma levels of 8-iso-PGF2αin two groups of patients. And NIHSS score was used to evaluate the severity of clinical neurological deficit. Results The plasma levels of 8-iso-PGF2α were significantly higher in old ACI group (506.38±138.63) ng/L than those of middle-young ACI group (420.18±132.72) ng/L and old control group (369.98±99.81) ng/L. There was no significant difference in plasma level of 8-iso-PGF2αbetween middle-young ACI group and old control group (F=9.272,P<0.05). The NIHSS score was significantly higher in old group (19.78±3.66) than that of middle-young group (17.73 ± 2.70, t=2.539,P<0.05). There was a positive correlation between plasma 8-iso-PGF2α level and NIHSS score in old group (r=0.504,P=0.001). Conclusion The oxidative stress plays an important role in the occurrence and de-velopment process of ACI in elderly patients. The earlier and reasonable antioxidant therapy plays a positive role to alleviate the clinical symptoms and promote the recovery of illness.

ObjectiveThe purpose of the present study was to discuss the method to avoid spillage by means of adding chemotherapeutic drugs into sealed transfusion bottles.Methods90 penicillin sealed bottles of the same batch number were randomly divided into the test group and the control group,each with 45 bottles.Standard method according to Basic Nursing was used to add chemotherapeutic drugs into transfusion bottles in the control group.Drugs were added into transfusion bottles under negative pressure in the test group.The operating time was measured,the spillage volume of puncture site was calculated and microbial detection of syringe was observed in the two groups.ResultsThere was a significant difference in the spillage volume of puncture site between the two groups (P<0.01),but no difference was seen in the operating time and microbial detection of syringe (P>0.05),ConclusionsThe spillage volume of puncture site was reduced significantly by means of adding chemotherapeutic drugs into transfusion bottles under negative pressure.This can decrease chemotherapeutic professional exposing risks and drug wastage.

OBJECTIVE:To provide reference for improving the service quality of outpatient pharmacy in hospital. METH-ODS:A questionnaire survey was conducted to investigate and analyze the patients’satisfaction and related influential factors to outpatient pharmacy in a third grade class-A hospital in Chengdu. RESULTS:Totally 165 questionnaires were sent out,and 150 were effectively received with effective recovery of 90.91%. The total score for patients’satisfaction was (44.67 ± 7.81) scores, and the rate of satisfaction was(81.22±14.19)%. The top three entries were“the will you choose to come to our hospital again if necessary”,“the notices about time and place for taking the medicine”and“the overall evaluation of the professional ethics of med-ical staff”,scored 4.38,4.25 and 4.25,respectively;the last three entries were“waiting time for taking medicine”,“the notices about how long it takes to take the medicine”and“service facilities and environmental facilities for drug taking”,scored 3.55, 3.63 and 3.95,respectively. The top three suggestions were“long waiting time for taking medicine and inconvenient”,“noisy envi-ronment,bad order”and“expensive drugs charges but less reimburse”. The univariate analysis and multivariate analysis showed that gender,age,marital status,education,occupation,family income per month,resident,drug taking times and payment etc. factors showed no significant effects on patients’satisfaction scores(P>0.05). CONCLUSIONS:The degree of patients’satisfac-tion in outpatient pharmacy have no obvious specificity and preference,the key to improve the degree of satisfaction lies on strengthening the service of the hospital and the perception of the patients. While the next research will focus on how to find the breakthrough points and key points to improve the experience of waiting,standardize process management and logistics manage-ment,and make patients aware of the service development.

To observe the proliferation of T lymphocytes stimulated by CML and AML cells which were induced by rhGM-CSF and rhIL-4, and the secretion of IFN-gamma from proliferated T lymphocytes, the expression of CD80, CD86 and HLA-DR on CML and AML cells induced by GM-CSF and IL-4 was assayed by flow cytometry in vitro. Then one-way mixed lymphocyte reaction was carried out, with induced leukemia cells as stimulating cells and auto-T lymphocytes as reactive cells. The secretion of IFN-gamma from T lymphocytes was determined by double antibody sandwich ELISA. The results showed that GM-CSF and IL-4 significantly upregulated the expression of CD80, CD86 and HLA-DR on CML cells and CD80 and CD86 on AML cells, which could stimulate the T lymphocyte proliferation and high secretion of IFN-gamma (in CML group) of autologous T lymphocytes. It is concluded that the CML and AML cells induced by GM-CSF and IL-4 have the ability to present tumor specific antigen to auto-T lymphocyte.
Adult ; Aged ; Aged, 80 and over ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interferon-gamma ; biosynthesis ; Interleukin-4 ; pharmacology ; Leukemia, Myeloid ; immunology ; Lymphocyte Activation ; drug effects ; Male ; Middle Aged ; Recombinant Proteins ; pharmacology ; T-Lymphocytes ; drug effects ; immunology

Objective To analyze the research status and hotspot of occupational methanol poisoning at home and abroad. Methods ， The China National Knowledge Resource Database Wanfang Data Knowledge Service Platform and Web of Science were used as the data sources. The relevant literatures on occupational methanol poisoning published in domestic and foreign ， Results journals up to June 30 2021 were searched. The bibliometrics was used to analyzed the literatures. A total of 255 literatures were included in analysis. There were 187 Chinese articles and 68 English articles. Most of Chinese articles were ， ， published from 2001 to 2005 with an average of 26.7 literatures per five years until June 2021. Among them 72 literatures （ ）， ， were published in core journals 38.5% and 176 authors from 27 provinces autonomous regions and municipality directly ， under the central government published relevant literatures. The research contents mainly focused on the classification ， ， poisoning mode clinical manifestations visual impairment and poisoning prevention and treatment of occupational methanol - ， poisoning. Most of the English literatures were published in 2016 2020 with an average of 4.9 articles per five years until June ， （ ）， 2021. Among them 36 were published in SCI journals 52.9% and 57 authors from 11 countries published relevant ， ， ， literatures. The research contents mainly focused on the clinical diagnosis drug treatment intoxication mechanism visual Conclusion sequelae and brain injury of occupational methanol poisoning. The research on occupational methanol poisoning ， ， ， mainly focuses on clinical diagnosis clinical manifestations treatment and prognosis and pathogenesis. The focus of relevant research at home and abroad is different.

A sensitive determination method for sulfur mustard ( HD ) metabolites thiodiglycol ( TDG ) in rabbit urine was established and validated using isotope dilution negative ion chemical ionization ( NICI) gas chromatography-mass spectrometry ( GC-MS ) , in which deuterated thiodiglycol ( TDG-d8 ) was used as internal standard. Two solid-phase extraction ( SPE) steps were established and optimized in order to reduce the interfering backgrounds, one was used to extract thiodiglycol ( TDG ) from urine with self-assemblied Florisil SPE cartridges, another cleaning treatment of the by-products after pentafluorobenzoyl chloride (PFBZ) derivatization. The results showed that the limits of detection quantitation of this method were 0. 1 and 0. 3 μg/L, respectively. The exposure time-response relationship and exposure dose-response relationship of TDG in rabbit urine were studied after rabbit skin exposure to sulfur mustard (HD, 0. 02-0. 15 LD50). The TDG levels in the rabbit urine increased rapidly during the first day after application and then decreased over time for all dosage groups. A secondary release was also noted for the high-dose group, and the duration of high TDG excretion levels was correlated positively with the HD dosage levels. We thus concluded that abnormally high levels of TDG in urine could be used as a clear diagnostic indicator of HD exposure.

BACKGROUND:Neuroblastoma is a common solid tumor in children. Pediatric tumors are little affected by environmental factors, but closely related to child development. The suspension method is an effective and reliable method to harvest neoplastic stem cel s.
OBJECTIVE:To culture the cel clones of human neuroblastoma cel line SK-N-SH and to assess the biological properties of the cel clones.
METHODS:Using the suspension method with no serum media, tumor cel clones were obtained. Immunofluorescence method was used to identify whether tumor cel clones exhibit stem cel properties. SK-N-SH neuroblastoma was labeled by luciferase, and tumor cel clones and tumor cel s were seeded onto the back of nude mice to monitor cel proliferative properties in nude mice using in vivo imaging.
RESULTS AND CONCLUSION:Using the suspension culture method, SK-N-SH neuroblastoma cel s could successful y develop into cloning bal s. Under serum-free culture, cloning bal s were immunofluorescently used to detect molecular markers that showed strong positive expression. Cloning bal s subcutaneously implanted into
nude mice showed the strong ability of self-renewal and differentiation as stem cel s. The cel clones cultured by the suspension method strongly expressed Nestin, but weakly expressed glial fibril ary acidic protein, neuron-specific tubulin, possessing stem cel characteristics and strong proliferation and metastasis in nude mice.

Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.

Objective To investigate the clinical effect and the prospect of Infliximab in treatment of intravenous immunoglobulin (IVIG)-resistant Kawasaki disease (KD) patients.Methods Clinical features,inflammatory markers and coronary changes were observed in 2 cases of IVIG-resistant KD patients hospitalized in Peking University First Hospital,who were treated effectively by Infliximab.Relevant researches on the mechanism and progress of the Infliximab treatment for IVIG-resistant KD in the last 10 years were reviewed at the same time.Results Two KD patients hospitalized in Peking University First Hospital had been treated with 2 g/kg IVIG for 2 times and followed by methylprednisolone treatment.However,fever and other clinical manifestations occurred again after 2 days and 6 days when temperature returned normal.They both defervesced and all the symptoms were improved after 1 dose of Infliximab (5 mg/kg) by laboratory examinations.Four published literatures of the basic research and 9 retrospective or prospective clinical researches of Infliximab treatment of KD showed that Infliximab alleviated the inflammatory level in the KD patients significantly.Complete remission was up to 72.73％-92.11％.Those KD patients defervesced within 12 h,with dramatic improvement of symptoms and signs.Arthralgia also disappeared in the patients with arthritis.Only 1 case was complicated with hepatitis in the acute phase and cholecystitis in recovery time.A phase 3 randomised,double-blinded,placebo-controlled trial had been done to assess the addition of Infliximab to the standard therapy.Conclusions Infliximab is a feasible choice for IVIG-resistant KD patients.Efficacy and safety of Infliximab for KD treatment have been proved in the literature.However,Infliximab for KD treatment has not been indicated in the drug instruction,so the informed consent from the guardians and Ethics Committee is needed.