With the in-depth study of related substances and the development of consistency evaluation of generic drugs, relative correction factors are gaining increasing attention. By analyzing the domestic and foreign literature on correction factors in recent years, this paper describes the correction factor component, the current measurement method and its application. The rules and key points of use of an impurity correction factor and its determination and application are described, and some problems in its determination and application are discussed, providing a reference and basis for the standardization of research on impurity correction factors in the future.

Research on polymer impurities has always been important in the quality control of cephalosporins. Research on polymers in cephalosporins that lack active amino groups on the C-7 side chain has not been reported. Therefore, our study used cefazolin sodium, which is widely used in the clinic, as an example. The polymer in cefazolin sodium and its product "cefazolin sodium pentahydrate for injection" was analyzed by column switching liquid chromatography-high resolution mass spectrometry. Two polymer impurity peaks were detected and the possible structures of these polymers were suggested. Through two-dimensional liquid chromatography, the chromatographic peaks following Sephadex gel chromatography and high-performance gel chromatography were compared to those obtained by reverse high-performance liquid chromatography (HPLC) for cefazolin sodium as reported in the Chinese Pharmacopoeia. The HPLC method proves more suitable for polymer detection than Sephadex gel chromatography and high-performance gel chromatography. The method of polymer detection for cefazolin sodium was established using the method of related substances HPLC as described in the Chinese Pharmacopoeia.

The quality of some earlier developed antibiotics is usually ensured by the combination of HPLC purity and microbiological potency measurement in the pharmacopoeias of various countries because the relationship between their purity and potency is not clearly quantified. Due to potency is assessed using certain units of measurement, it can not be directly traced to the international system of units (SI unit). This has become a hotspot in the study of the quantitative relationship between purity and potency of antibiotics. It would be quite an achievement to simultaneously determine both purity and potency using HPLC methods during quality control. This study evaluated a multicomponent antibiotic product, gentamycin, as a test sample. First, pure samples of the C components of gentamycin: C1a, C2, C2a and C1 were prepared, separately. Second, quantitative relationship (theoretical potency) between the purity and potency of each C component of gentamycin were determined using 1H NMR, HPLC-ELSD and microbiological assay method. One milligram of gentamycin C1a, C2, C2a and C1 was equal to 1 286.98, 1 095.74, 1 079.52 and 739.61 gentamycin units, respectively. Finally, a method for the determination of gentamycin potency was established based on the proportion and content of C components of gentamycin. The unification of purity and potency for gentamycin was achieved using only HPLC-ELSD. It is also demonstrated that C components of gentamycin and micronomicin produce the same responses under ELSD, which means that it is not necessary to prepare separate reference standards for each C component of gentamycin and that quantitative testing can be performed accurately using only one micronomicin reference standard. This study simplified the previous method for the determination of the content of C components of gentamycin using HPLC-ELSD. The developed method is suitable for regular use as a part of quality control and can simplify the rigmarole quality control procedures provided in current pharmacopeias.
Chromatography, High Pressure Liquid ; Gentamicins ; chemistry ; pharmacology ; Magnetic Resonance Spectroscopy ; Microbial Sensitivity Tests ; methods ; Molecular Structure ; Quality Control ; Reference Standards

OBJECTIVETo investigate the incidence of nosocomial infections of newborn infants in neonates and to explore the risk factors and strategies of infection control.

METHODSThere were 433 confirmed cases of nosocomial infection in the neonatal ward of the authors' hospital from January 2007 to December 2009. Their data of epidemiological and clinical characteristics, results of etiological examinations and antibiotic resistance were retrospectively analyzed.

RESULTSDuring the study, the number of hospitalizations were 6437. Nosocomial infection occurred in 433 patients 513 times. The overall nosocomial infection rate was 6.82%. The overall hospitalization days were 73 663 and nosocomial infection patient-day rates were 6.96‰. The VAP infection rate was 28.7‰. The CRBSI rate was 3.5‰. Gestational age (OR = 1.049), mechanical ventilation (OR = 1.810), umbilical vein catheter (OR = 1.106), hospitalization days (OR = 1.081), premature rupture of membrane (OR = 1.433) were the risk factors for the development of nosocomial infection. There were 197 (38.4%) cases of pneumonia, which was the most common nosocomial infection in Neonatal Ward. There were 129 cases of ventilator-associated pneumonia (VAP), which accounts for 65.5% of pneumonia and 24.4% of cases treated with ventilator. The next was sepsis, 124 cases (24.2%) and 64 cases of diarrheal disease (12.7%). One hundred and eighty two (54.4%) strains of isolates were Gram-negative bacteria, which accounted for the highest proportion. The predominant pathogens of Gram-negative bacteria were Klebsiella pneumoniae (19.6%), followed by Acinetobacter baumannii (8.1%), Pseudomonas aeruginosa (7.2%), Stenotrophomonas maltophilia (4.8%) and Escherichia coli (4.8%). The isolation rates of Klebsiella pneumoniae and Escherichia coli with positive extended-spectrum beta-lactamases (ESBLs) were 91.4% and 75%, respectively. Those two bacteria were universally resistant to cephalosporins. The rate of resistance to imipenem of Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa were 1.5%, 11.1% and 41.7%. The isolation rates of methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative Staphylococcus were 28.6% and 95.5%.

CONCLUSIONIt is important to identify the high risk factors for nosocomial infections in newborn infants. To shorten time for mechanical ventilation and hospitalization days, removal of the central venous catheter as early as possible would be conducive to reducing the morbidity of nosocomial infection. The main pathogens were Gram-negative bacteria. The multidrug resistance of Enterobacteriaceae and Non-fermenters is serious.

Cross Infection ; epidemiology ; microbiology ; Female ; Humans ; Incidence ; Infant, Newborn ; Intensive Care Units, Neonatal ; Retrospective Studies ; Risk Factors

0.05 for all).Conclusions The allelic frequencies of LPL gene at Pvu Ⅱ locus in Hei Yi Zhuang were different from those in Han,but the genotypie frequencies in Hei Yi Zhuang were not different from those in Han.There was no significant correlation between the polymorphism of LPL gene at Pvu Ⅱ site and the serum lipid levels in two ethnic groups.

AIM: To observe the efficacy of surgical excision combined with autologous limbus conjunctival flap transplantation in the treatment of pterygium accompanied with conjunctival cyst.METHODS: Totally 126 patients 188 eyes with pterygium were hospitalized in Department of Ophthalmology of Tongji Hospital of Huazhong University of Science and Technology during August 2013 and August 2015.The patients were divided into two groups: observation group (11 eyes of 11 patients) with pterygium accompanied with conjunctival cyst and control group (177 eyes of 115 patients) with primary pterygium.All patients underwent slit lamp microscope examination, anterior segment photography, and anterior segment optical coherence tomography(OCT).The size of pterygium was calculated by multiplying neck width and length of the covered corneal.All patients underwent excision combined with autologous conjunctival flap transplantation, and the resections were performed pathological section with hematoxylin and eosin staining.All patients were followed up postoperatively for 4-28mo.RESULTS: All cases in the observation group were confirmed by postoperative pathological examination.All cyst walls were complete, and containing single layer of epithelial cells.The mean size of pterygium of the observation group was 6.9±1.7mm2, and 6.3±1.8mm2 for the control group.There was no significant difference between the two groups (P>0.05).The mean postoperative healing time of observation group was 2.1±0.9d, and 1.9±0.8d for the control group.There was no significant difference between the two groups (P>0.05).Recurrence was seen in two eyes within the follow-up period in the control group, and no recurrence in the observation group.CONCLUSION: Surgical excision combined with autologous limbus conjunctival flap transplantation is a safe and effective treatment for pterygium accompanied with conjunctival cyst.

OBJECTIVETo investigate molecular epidemiologic features of rotavirus (RV) infection in infantile diarrhea in Hangzhou area.

METHODSStool specimens of 683 infants with suspected acute viral enteritis in the autumn and winter of 2001 - 2003 were collected. RV (group A) was detected by using latex agglutination test (LAT). VP7 serotype (G) positive specimens were detected by using enzyme linked immunosorbent assay (ELISA) and then the RNA of the virus was determined with reverse transcription polymerase chain reaction (RT-PCR). cDNA of VP7 gene fragment was sequenced by automatic gene analyzor (ABI3730) and compared with the RV VP7 gene sequences stored in Genebank.

RESULTSRV was detected in 297 of 683 (43.5%) specimens by LAT. The highest frequency of RV (group A) detected was 52.9% (228/431) in patients aged 7 - 18 months. The prevalent serotypes were G1 (36.7%, 109/297) and G3 (30.9%, 92/297), followed by mixed type (11.8%, 35/297), untyped (9.4%, 28/297), G4 (7.1%, 21/297) and G2 (4.0%, 12/297). The prevalent serotypes seen each year were different. G1 (54.9%, 45/82) was the major serotype in 2001 followed by G3 (14.6%, 12/82). In 2003, the major serotype was G3 (43.0%, 63/146) and followed by G1 (29.5%, 43/146). The reliability of ELISA was confirmed by RT-PCR, gene sequencing and homology analysis.

CONCLUSIONThe main prevalent serotypes of VP7 of rotavirus were G1 and G3. The dominant serotypes of rotavirus varied in Hangzhou area from 2001 to 2003.

Antigens, Viral ; classification ; genetics ; metabolism ; Capsid Proteins ; classification ; genetics ; metabolism ; Child, Preschool ; China ; epidemiology ; Diarrhea, Infantile ; epidemiology ; virology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Infant ; Latex Fixation Tests ; Male ; Molecular Sequence Data ; Prevalence ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; classification ; genetics ; Rotavirus Infections ; complications ; epidemiology ; virology ; Serotyping

This study is to investigate the effect of diet and gene on blood-fat trait of the individual mouse. One hundred and twenty mice were fed with high fat emulsion for 4 weeks. Then the genotypes of Glut-4 were analyzed by PCR-SSCP to investigate the effect on blood-fat traits and some organ performance of mice. After fed with high fat emulsion for 4 weeks, 98 mice suffered from hyperlipidemia, while 22 mice did not. And a T/A mutation was found in 188 targeted fragments we amplified at 8,521 site, and association analysis showed that the mice carried BB genotype had higher TC, TG and LW/BW level than those of the mice with AA genotype (P < 0.05). Different Glut-4 genotypes show different sensitivities to high fat emulsion on mice.
Animals ; Glucose Transporter Type 4 ; genetics ; Hyperglycemia ; blood ; genetics ; Lipids ; blood ; Mice ; Mice, Obese ; genetics ; Polymorphism, Single Nucleotide

Aim To assess the effects of Banqiao Codonopisis Pilosula(BCP)decoction on learning and memory dysfunction in AD model rats induced by high activity GSK-3β and its possible mechanism.Methods The SD rats(4 months old,♂)were divided into five groups,namely,sham-operated group(blank group),AD model group,BCP high-dose(2.16 g·kg-1·d-1)group,BCP medium-dose(1.08 g·kg-1·d-1)group,and BCP lower-dose(0.54 g·kg-1·d-1)group.Treatment group received BCP decoction by gavage once a day for 14 days,while other groups were offered drinking water by gavage once a day for 14 days.The autonomous behavior activities of all rats were observed and recorded after gavage.In the last seven days by gavage,Morris water maze test was used to test the spatial learning and memory ability of the five groups.After five days training,treatment groups and AD model group were injected wortmannin(WT,PI3K specific inhibitor)and GF-109203X(GFX,PKC specific inhibitor)(100 μmol·L-1 of each,total volume of 10 μL)into the right lateral ventricle of the rats.The blank group was only injected 2％DMSO.The spatial memory retention was detected by water maze 24 hours after lateral ventricle injection.Then,changes in the spatial learning memory of rats were observed.The level of Tau phosphorylation in SD rat hippocampus and the expression and activity changes of related protein kinase GSK-3β were detected by Western blot and immunohistochemistry.The changes of Nissl bodies in SD rat hippocampus were observed by Nissl′s staining.Results After intragastric administration of BCP,the rat autonomous behavior activities in each group all showed a declining trend,and the differences in low-dose and middle-dose groups had statistical significance compared with blank group.The Morris water maze tests showed that the latency navigation of model group was significantly longer than that of blank group(P<0.01),while that of the BCP three doses groups was shorter than that of model group(P<0.05).Compared with the same group,the latency navigation of the three groups after gavage BCP low,middle and high dose was significant shorter than that without gavage(P<0.05).Western blot results showed that the activity of GSK-3β in AD model group was up-regulated compared with the blank group.However,BCP inhibited activity of GSK-3β.Western blot and immunohistochemistry results showed the level of Tau phosphorylation in AD model group was increased compared with the blank group in the area of CA3(P<0.05).Compared with AD model group,the level of Tau phosphorylation was decreased in treatment group.Nissl′s staining results showed that dendritic spines in AD model group was significantly attenuated compared with the blank group(P<0.05).Far more dendritic spines were observed in treatment group than in AD model group.The number of Nissl′s bodies in neuron cells of hippocampus in hippocampal CA3 was obviously larger in treatment groups than in AD model group.These effect of BCP was dose-dependent.Conclusions BCP can prevent the learning and memory dysfunction in AD model rats induced by high activity of GSK-3β.The mechanism may be related to inhibiting GSK-3β activity and then reducing the level of phosphorylation of Tau and improving neural development.

The natural isoflavones biosynthetic pathway is only limited in legumes plant. To study the isoflavone in bacteria by metabolic engineering requires transformation of multi-gene of the whole pathway into the host strain to resembling the expression and metabolism of the genes. The multi-gene transformation and expression strategy become necessary because of this. This article talks about the multi-gene transformation strategy using one or many vectors, taking the five genes of isoflavonoid biosynthetic pathway to E. coli. The recombinant bacteria carry five genes with two vectors, the whole Isoflavonoid biosynthetic pathway was constructed into E. coli. Fermented with L-tyrosine as substrate and IPTG as an inducer, the recombinant bacteria can produce a new isoflavone related metabolite showed on HPLC analysis profile.
Amino Acid Sequence ; Base Sequence ; Biosynthetic Pathways ; genetics ; Enzymes ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Engineering ; methods ; Isoflavones ; biosynthesis ; metabolism ; Molecular Sequence Data ; Oxygenases ; biosynthesis ; genetics ; Soybeans ; chemistry ; Transformation, Bacterial