Objective:To investigate the ways of pharmaceutical care performed by clinical pharmacists for the patients with severe infection. Methods:Through deciding the anti-infection therapeutic regimen, providing drug counselling and pharmacy education and focusing on adverse drug reactions, pharmacists offered suggestions for one patient with severe legionella pneumonia complicated with AECOPD. Results:The pharmaceutical care performed by clinical pharmacists could solve the problems and improve the compliance, safety, effectiveness and rationality in the drug treatment of the patient. Conclusion:According to the individual condition of patients, clinical pharmacists can realize their own values through looking for the breakthrough points of pharmaceutical care and participating in clinical practice.

Objective To construct the eukaryotic expression vectors of fibroblast growth factor receptor 3(FGFR3) MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN, and to detect their expressions in human chronic myeloid leukemia(CML)K562 cell line.Methods The full-length FGFR3 (fgfr3-WT)and dominant negative FGFR3 (fgfr3-DN)were amplified by polymerase chain reaction (PCR). The two genes were respectively digested with EcoRⅠand BamHⅠ,and then ligated into MSCV/puro to construct the recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN which were tranduced into K562 cells by LipofectaminTM 2000 after PCR,double digestion and DNA sequencing.The expressions of FGFR3 protein in K562 cells were detected by Western blotting and flow cytometry. Results The recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN were amplified by PCR method, and the results showed fgfr3-WT of 2 400 bp and fgfr3-DN of 1 200 bp had been successfully cloned into MSCV-puro vector. The 2 400 bp fragment was oblained after double digestion of recombinant plasmid.The sequencing results showed that the size of fgfr3-WT was 2 400 bp which was the same as the sequence from GeneBank.Fgfr3-DN of 1 200 bp was also in conformity with the expected sequence.Compared with control (K562 MSCV)group,the expression level of FGFR3-WT in MSCV/puro-fgfr3-WT transfection (K562-WT)group was increased to above 10 times.There was high expression of FGFR3-DN in MSCV/puro-FGFR3-DN transfection (K562-DN)group,but there was no expressions in control(K562 MSCV)group and K562-WT group.The flow cytometry results showed that the high expressions of FGFR3-WT were in 57.5% cells in K562-DN group and the high expressions of FGFR3-DN were in 41.5% cells in K562-DN group. Conclusion The K562 cell lines highly expressing FGFR3-WT and FGFR3-DN are constructed successfully.

Objective To evaluate the efficacy and safety of CT-guided 125I radioactive seed implantation combined with gemcitabine and Gio (gemcitabine and S-1, GS scheme) chemotherapy in treating advanced pancreatic carcinoma. Methods Sixty-eight patients with inoperable advanced pancreatic carcinoma were randomly divided into two groups. Patients in group A(n=38) were treated with CT-guided 125I radioactive seed implantation combined with GS chemotherapy scheme, while patients in group B (n=30) received GS chemotherapy scheme only. The short-term effect, the median progression-free survival time, the median survival time and adverse reactions of the two groups were determined , and the results were compared between the two groups. Results The objective response rate (ORR), disease control rate (DCR) and clinical benefit rate (CBR) of the group A were 57.9%, 73.7%and 84.2%respectively, while those of group B were 26.7%, 46.7% and 60.0% respectively. The differences between the two groups were statistically significant (P<0.05). In group A the median progression-free survival time and the median survival time were 8.00 months and 11.84 months respectively, which were strikingly higher than those in group B (5.63 months and 10.40 months respectively), the differences between the two groups were statistically significantly (P<0.05). No significant differences in gastrointestinal reactions, blood toxicity, liver toxicity and other adverse reactions existed between the two groups (P>0.05). Conclusion For advanced pancreatic carcinoma, CT-guided 125I radioactive seed implantation combined with GS program is a safe and effective treatment.

Objective To investigate the value of CT-guided 125I particles implantation combined with gemcitabine plus S-1 (GS) regimen in the treatment of locally advanced pancreatic cancer.Methods 42 patients with unresectable local advanced pancreatic cancer were given with CT-guided 125I seed implantation.3-4 cycles of GS regimen was given based on the tolerance of patient s body within 3-7 d after implantation of particles.Review of blood,CA199,chest X-ray,CT scan + enhanced or MRI were performed at 2nd,4th,6th,12th month after surgery.Results 2nd,4th,6th month after surgery,tumor lesions were significantly reduced,ORRs were 59.5 ％ (25/42),66.7 ％ (28/42) and 73.8 ％ (31/42),respectively,DCRs were 83.3 ％ (35/42),78.6 ％ (33/42) and 76.2 ％ (32/42),respectively.No serious adverse reactions were observed,patient could tolerate these reactions.The 6th,12th,24th month survival rates were 100 ％ (42/42),47.6 ％ (20/42) and 11.9 ％ (5/42),respectively,mPFS was 8.27 months and mOS was 12.00 months.Conclusion CT-guided 125I particles implantation combined with GS regimen is convenient,safe,high efficacy in the treatment of locally advanced pancreatic cancer.

Objective: To establish the quality assessment of Cordyceps and its cultured Cmycelia by TLC fingerprint, and to compare the content of the nucleotides between them. Methods: Adenosine, Uridine and Guannosine were detemined by TLC-Scanning samples were perfomed by fluorescence quenching analyis. Results: The contents of adenosiue, uridine, and guanosine from cordyceps mycelia were more than that of cordyceps from wildlife. Conclusion: This method is convenient and rapid, and it can effectively evaluate nucleotides in nature Cordyceps and culture Cordyceps.

Objective To improve differential diagnosis of tumefactive demyelinating lesions (TDL) and primary central nervous system lymphoma (PCNSL) by analyzing the clinicopathological features of the diseases.Methods The clinical features,neuroimaging findings and pathological characteristics of 4 patients with pathologically proven TDL and 9 patients with pathologically proven PCNSL were retrospectively analyzed.Computer tomography and magnetic resonance imaging were used for neuroimaging studies.The hematoxylin and eosin staining,Luxol Fast Blue staining and immunohistochemistry were used for pathological studies.Results (1) The features of lesions on brain imaging scan:CT in TDL patients showed low density.Enhanced MRI demonstrations were different in different courses:3 cases with ring enhancement,1 case with spotty strengthen;5 PCNSL cases showed hyperdensity in CT,1 case showed isodensity,and 3 cases low-density.MRI showed enhancement of uniform enhancement in PCNSL patients.(2) The features of lesions on pathology:the plaques of lesions in TDL patients were characterized by massive demyelination with relatively axonal preservation associated with prominent astrocytosis and profound infiltrates composed.Typical pathological features in PCNSL cases were that tumor cells around blood vessels showed the cuff-like arrangement.Due to use of hormones and other causes,pathological demonstrations of a part of PCNSL cases were atypical,which were easily confused with TDL.There were 4 cases with more than one biopsy for diagnosis.Conclusions (1) PCNSL with low or equal density in CT needs to be differentiated with TDL.(2) The pathological features of some cases of PCNSL after hormone therapy were similar to TDL.It is better not to use hormone before definite diagnosis.(3) The pathology of PCNSL may be related to the progression of the disease.Some of patients need to be re-biopsied.It is important to combine clinical imaging and pathology for diagnosis of the disease,and attention should be paid to followup.

BACKGROUND:Nano-hydroxyapatite as a surface modification material that is bonded to the surface of the zirconia ceramics upon sintering at high temperature can improve bone-inducing activity and bone bonding strength of the zirconia ceramics. Moreover, the sintering temperature is crucial for performance and bonding of the composite. OBJECTIVE:To detect the shear strength of nano-hydroxyapatite ceramics coating bonded to zirconia ceramics at different sintering temperatures. METHODS:Nano-hydroxyapatite slurry was prepared using sol/gel technology. Thereafter, 20 zirconium green bodies were coated with nano-hydroxyapatite slurry and randomly divided into four groups. Then, the specimens were put into non-pressure sintering furnace and sintered at 1 300, 1 400, 1 500, and 1 550℃, respectively. At last, we measured the shear strength of al the specimens after sintering by universal testing machine, and analyze the type of fractures. RESULTS AND CONCLUSION: With the rising of sintering temperature, the shear strength of the specimens was gradualy increased, and there were significant differences between the four groups [(4.04±1.19), (6.60±0.95), (16.51±1.93), (80.47±19.31) MPa,P < 0.05]. Within the scope of 1 550℃, the sintering temperature was positively relative to the shear strength of specimens. These findings indicate that in the certain temperature range, the higher the sintering temperature, the greater the shear strength of the bonding interface between zirconia and nano-hydroxyapatite. When the sintering temperature is 1 550℃, the shear strength of the bonding interface is the highest.

To evaluate the virulence genes and antibiotics resistance of Staphylococci species in mastitis cows isolated from parts of Guangdong,and then provide scientific basis of the prevention of bovine mastitis.Forty strains Staphylococci isolated from milk samples of 110 mastitis cows were collected,and virulence genes,resistance genes and disinfectant resistant genes were detected by PCR,and antibiotics resistance with K-B paper method.The results showed that the highest virulence gene was fnbp (17.5％),followed by seb (15％) and tsst (15％),and virulence genotype was complex.The most prevalent antibiotic resistance drug wvas streptomycin,followed by erythromycin and penicillin G,and CNS were susceptible to oxacillin,ceftraxone and cefazolin.The most prevalent antibiotic resistance genes was quinolones gene qnrA/B/C/D(20％),the resistance to streptomycin was mediated by aac6-aph2.The most prevalent disinfectant resistant gene was qacG (225 ％).The genotype of antibiotics resistance gene and disinfectant resistant gene was complex.

Objective:To establish the method for detecting lower respiratory infections (LRIs) bacterialpathogens using nanopore sequencing, and evaluate the feasibility of this method.Methods:Bronchoalveolar lavage fluid (BALF) samples from 33 patients with LRIs who visited the Department of Respiratory and Critical Care Medicine of Beijing Hospital from July 2019 to September 2020 were collected.Nanopore 16S amplicon sequencing were performed on these samples. In order to evaluate the clinical value of the nanopore sequencing, χ 2 test was used to analyze the pathogen differences between the detection rate and pathogen types results found with using the nanopore 16S sequencing and the results found with bacterial culture. Results:The process and method of nanopore sequencing used in the detection of the LRIs pathogens were established. The pathogen detection rate of the 16S sequencing was higher than that of the traditional bacterial culture (75.8% [25/33], 45.5% [15/33], χ2=5.140, P<0.05). From the 25 positive samples found with nanopore 16S sequencing, 16 pathogens were detected, including Haemophilus parainfluenzae, Haemophilus influenzae, Streptococcus pneumoniae, Streptomonas maltophilia, Acinetobacter baumannii, and Acinetobacter junii, Staphylococcus aureus, Klebsiella pneumoniae, Enterococcus faecalis, Enterococcus gallinarum, Corynebacterium striatum, Mycobacterium paraintracellulare, Serratia marcescens, Achromobacter insuavis, Citrobacter murliniae and Mycoplasma pneumoniae. More than 6 pathogens were tested in clinical culture, including Haemophilus parainfluenzae, Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and Streptomonas maltophilia (χ2=7.949, P<0.05). 16S sequencing aligned to species level sequences accounted for 80.0 (60.0, 86.0)% of the genus level. The results obtained by using16S sequencing and bacterial culture were consistent in 11 (33.3%) samples. Conclusions:Nanopore 16S amplicon sequencing can quickly identify pathogenic bacteria from BALF in LRIs patients. Nanopore 16S amplicon sequencing has a high detection rate, it can detect more pathogens than traditional bacterial culture, and it can also identify most bacteria to the species level. This technology is a very promising platform with broad application prospects.

The methylation status of GNAS1 gene in pseudohypoparathyroidism type Ib patients was detected by methylation-specific PGR technique. There was an abnormal methylation of 1A region in all seven PHPIb patients. Loss of exon 1A methylation (imprinting defect) seems to be the cause of PHPIb.