To establish a UPLC-MS/MS method for the simultaneous determination of geniposide, genipin 1-O-beta-D-gentiobioside and geniposidic acid in rat brains and study the brain pharmacokinetics of the three iridoid glycosides in stroke rat after the oral administration of Xingnaojing. In this experiment, brain samples were precipitated with protein for twice. Acquity BEH C18 column was adopted, with acetonitrile-0.1% formic acid-water as the mobile phase for gradient elution. ESI source was adopted for mass spectra; multiple reaction monitoring (MRM) was conducted to detect negative ions. The time for sample analysis was 3.5 min. the results showed good linear relations among the three iridoid glycosides, with the extraction recovery between 99.6% and 114.3%, good intra- and inter-day precisions and accuracies and stability in line with the requirements. The t1/2 and MRT in the three components were similar in brains of stroke rats. Geniposide and genipin 1-O-beta-D-gentiobioside showed double peaks; where as geniposidic acid showed a single peak. In conclusion, the method is so specific, sensitive, accurate and reliable that it can be used to study the brain pharmacokinetics of Xingnaojing oral preparation.
Animals ; Brain ; metabolism ; Brain Chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Iridoids ; chemistry ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Objective To study the mini-invasive surgical treatment for cholelithiasis disease in the primary hospital. Methods Laparoseope,duodenoscope and choledochoscope were combined to treat 293 cases with cholelith-iasis which included 2 cases cholecystolithisasis, 131 cases were cholecystolithiasis with secondary choledocholith,28 cases with primary choledocholith,39 cases with calculus of intrahepatic duct,93 cases with calculus of intrahepatic or extrahepatic duct. Results The program had minor trauma, mild suffering, and less operative complications,compared with traditional open operations. Conclusion The combined treatment of laparoscopy, duodenoscopy, choledochoscope and surgery for the cholelithiasis in the primary hospital is feasible, safe and effective.

Objective To study the effects of Yiqi Huayu Jiedu Prescription on the migration ability of MHCC97-H and the expressions of CXCL12, CXCR4 and CXCR7; To discuss its relevant mechanism of action. Methods Setting Sorafenib as a positive control, CCK-8 method was used for determining the effects of Yiqi Huayu Jiedu Prescription on the cell proliferation of MHCC97-H and the optimum concentration. Scratch assay was used to observe the migration ability of MHCC97-H. The protein expressions of CXCL12, CXCR4 and CXCR7 were detected by Western blot after 24 h of medicine intervention. Results Yiqi Huayu Jiedu Prescription and Sorafenib can inhibit the cell proliferation of MHCC97-H , and the inhibitory concentration was 0.095 g/mL and 10 μmol/mL at 24 hours. Yiqi Huayu Jiedu Prescription can inhibit migration ability of MHCC97-H. The protein expressions of CXCL12, CXCR4 and CXCR7 in hepatocellular carcinoma cells decreased after the action of Yiqi Huayu Jiedu Prescription. Conclusion Yiqi Huayu Jiedu Prescription can inhibit MHCC97-H cell proliferation and migration, which may be realized by down-regulating chemokine axis of CXCL12/CXCR4/CXCR7.

OBJECTIVETo study the existence of Ehrluichiosis, lyme disease and tick-borne spotted fever coinfection in some areas in China.

METHODSUsing polymerase chain reaction (PCR), B. burgdorferi sensu lato, spotted fever group (SFG) Rickettsiae and human granulocytic ehrlichia (HGE), Ehrlichia chaffeensis (EC) were detected in ticks and mouse samples collected from Inner Mogolia autonomous region, Heilongjiang province, Beijing and Fujian province.

RESULTS408 Ixodes persulcatus collected from Inner Mogolia autonomous region, HGE and B. burgdorferi sensu lato and SFG Rickettsiae were detected positive, with rates as 6.8%, 7.8%, 45.6%, respectively. 5 (5/408) were coinfection with HGE and B. burgdorferi sensu lato while 1 (1/408) was coinfection with HGE and SFG Rickettsiae. 46 Ixodes persulcatus collected from Helongjiang province were determined positive, with rates as 6.5%, 10.8% and 34.8%, respectively including 1 (1/46) coinfected with HGE and B. burgdorferi sensu lato. 2 of 922 ticks collected from Beijing were detected positive with B. burgdorferi sensu lato. Among 283 groups of Haemaphysalis yeni ticks (3/group) and from 38 rodent samples collected from Ninghua county of Fujian province HCE and B. burgdorferi sensu lato and SFG Rickettsiae were detected. Out of them, 25 groups were positive with EC and the minimal positive rate was 3.8% while 21 rodent samples were positive with EC with a positive rate of 56.4%. 2 ticks and 1 rodent sample were detected positive with EC and spotted fever group.

CONCLUSIONCoinfection of HGE and B. burgdorferi sensu lato or spotted fever group Richi did exist in Ixodes persulcatus collected from Inner Mogolia autonomous region and Heilongjiang province. Coinfection of EC and spotted fever group Richi was found in the ticks and rodents collected from Fujian province.

Animals ; Arachnid Vectors ; Borrelia burgdorferi Group ; isolation & purification ; China ; epidemiology ; DNA, Bacterial ; analysis ; Disease Vectors ; Ehrlichia ; isolation & purification ; Ehrlichiosis ; epidemiology ; Humans ; Ixodes ; microbiology ; Lyme Disease ; epidemiology ; Polymerase Chain Reaction ; Rats ; Rickettsia ; isolation & purification ; Rickettsia Infections ; epidemiology ; Rodentia ; microbiology ; Tick-Borne Diseases ; epidemiology ; Ticks ; microbiology

OBJECTIVETo detect and study the types of Borrelia burgdorferi sensu lato in ticks and rodents from Da Xing-An Mountains Forest areas of China.

METHODSNested PCR was performed to amplify 5S-23S rRNA intergenic spacer of B. burgdorferi. Positive products were analysed by restriction fragment length polymorphism (RFLP) and single strand conformation polymorphism (SSCP), specimens showing unique RFLP profile were sequenced and analysed.

RESULTS1336 Ixodes persulcatus, 144 Dermacento silvarum, 144 Haemaphysalis concinna and 145 rodents of 9 species were collected from 16 sections of Da Xing-An Mountains Forest areas of China. Specific fragments were amplified from 293 I. persulcatus and 6 D. silvarum and 5 rodents of 4 species. B. burgdorferi was not detected in H. concinna. Among the positively tested I. persulcatus, 209 contained B. garinii genospecies and 45 contained B.afzelii genospecies based on RFLP. Moreover, B.garinii genospecies consisted of B. garinii 20047 and B. garinii NT29. 17 adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. garinii NT29. Nine adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. afzelii. Four adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. garinii NT29 and B. afzelii. Two D. silvarum were infected with B. garinii 20047, 1 D. silvarum with B. garinii 20047, 2 D. silvarum with B. afzelii. 3 rodents were infected with B. garinii 20047 while 2 rodents were infected with B. garinii NT29. Mixed infection was not found in D. silvarum and rodents. In addition, nine I. persulcatus and one D. silvarum specimens showed unique RFLP pattern. Data from sequential analysis showed that they all belonged to B. garinii. PCR-SSCP profiles of 5S-23S rRNA intergenic spacer of B. burgdorferi in the positive specimens exceeded 36 types; B. garinii 20047 showed 16 types while B. garinii NT29 showing 11 types, B. afzelii showing 9 types. SSCP profiles of the specimens coinfected with multiple B. burgdorferi was relatively complex.

CONCLUSIONThe infection of B. burgdorferi was found in the ticks and rodents in Da Xing-An Mountains Forests areas. The infection rate of I. persulcatus was high. B. garinii was predominant genospecies, and the population of B. burgdorferi was heterogeneous in the area. Mixed infections of different B. burgdorferi genospecies in ticks were found. I. persulcatus and Clethrionomys rufocanus were possibly served as major vector and major host for B. burgdorferi, respectively, suggesting that further study is needed to confirm the coinfection in humans and animals in this region.

Animals ; Borrelia burgdorferi Group ; genetics ; isolation & purification ; China ; epidemiology ; Humans ; Lyme Disease ; epidemiology ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational ; RNA, Bacterial ; analysis ; Rodentia ; microbiology ; Ticks ; microbiology ; Trees

OBJECTIVETo investigate rodents' natural infection of Orientia tsutsugamushi (Ot) in some areas of Inner Mongolia and Xinjiang, China.

METHODSDNAs were extracted from spleens of the captured mice and nested-polymerase chain reaction (nPCR) technique was used to detect the Ot-Sta56 gene. Six positive samples were sequenced and analyzed by Clustal X (5.0) and DNA Club software.

RESULTSA total of 90 rodents were captured in Inner Mongolia, and the overall prevalence of Ot was 6.67%. There was no significant difference in infection rates among the positive rodents species. 20 rodents were captured in Xinjiang, and the prevalence of Ot was 5.00%. The geographical difference in infection rates was not statistically significant between Inner Mongolia and Xinjiang. 9 rodents were captured in farmlands of Inner Mongolia and Xinjiang but there was no positive samples found. 101 rodents were captured in grasslands, and the prevalence of Ot was 6.93%. The Sta56 gene nucleotide sequence homology to Karp strain of N59 (from Microtus maximowiczii), N69 (from Cricetulus barabensis) and X33(from Cricetus cricetus) was 99%. The sequence homology to Taitung-2 strain and TW461 strain of N65 (from C. barabensis) was 94%, and the sequence homology to Taitung-2 strain and TW461 strain of N88(from Apodemus agrarius) was also 94%. The sequence homology to Oishi strain of N90 (from A. agrarius) was 96.00%.

CONCLUSIONOur findings indicated that infections of Ot did exist in rodents captured from Inner Mongolia and Xinjiang. The genotypes of Ot in Inner Mongolia and Xinjiang were quite complex, with some of them belonged to Karp type, and the others belonged to Taitung-2, TW461 and Oishi types which providing evidence for further investigation on the scrub typhus fuci in the two areas.

Animals ; China ; Geography ; Orientia tsutsugamushi ; genetics ; isolation & purification ; Polymerase Chain Reaction ; Rodentia ; microbiology ; Scrub Typhus

OBJECTIVETo investigate the efficacy of 2-month course of sleeping position correction in the treatment of positional plagiocephaly in infants aged <8 months.

METHODSA total of 73 infants with positional plagiocephaly between January 2015 and June 2016 were divided into treatment group (n=46) and control group (n=27) according to parents' wishes. The treatment group received sleeping position correction, while the control group received sleep curve mattress. The oblique diameters A and B in the two groups were measured and the cranial vault asymmetry (CVA) was calculated before and after treatment. The severity of positional plagiocephaly based on CVA was compared between the two groups before and after treatment. The Gesell Developmental Scale was used to determine the developmental quotients (DQs) in the motor, adaptive, language, and social domains in the two groups before and after treatment.

RESULTSBefore treatment, there were no significant differences in oblique diameters A and B, CVA, and DQs in the four specific domains between the two groups (P>0.05). After 2 months of treatment, the treatment group had a significantly greater oblique diameter B and a significantly smaller CVA than the control group (P<0.05); there were no significant differences in DQs in the four specific domains between the two groups (P>0.05). After treatment, both groups had significant improvements in oblique diameters A and B, CVA, and DQs in the motor and adaptive domains (P<0.01); moreover, the treatment group showed a significant improvement in the DQs in the social domain (P<0.01). There was no significant difference in the severity of positional plagiocephaly between the two groups before and after treatment (P>0.05).

CONCLUSIONSFor infants with positional plagiocephaly, sleeping position correction has better efficacy and is more convenient and economical than the sleep curve mattress, so it holds promise for clinical application.

Female ; Humans ; Infant ; Male ; Plagiocephaly, Nonsynostotic ; etiology ; therapy ; Posture ; Severity of Illness Index ; Sleep

OBJECTIVETo investigate the association between p53 Arg72Pro polymorphism and cervical carcinomas HPV-associated cervical carcinoma in Uigur and Han women.

METHODSThe distribution and frequencies of p53 Arg72Pro genotypes were determined by PCR-RFLP in 152 cases of cervical carcinoma in ethnic Uigur women with 110 cases of normal control and 120 cases of cervical carcinoma in Han women with 122 cases of normal control.

RESULTSThe omni-constituent ratio of p53 genotype was statistically different between cervical carcinoma and normal control groups in the Uigur (chi(2) = 7.196, P < 0.05) group. The proportion of Arg/Arg was higher in cervical carcinomas than that in control. The omni-constituent ratio of p53 genotype was statistically different between cervical carcinoma and normal control groups in Han (chi(2) = 8.231, P < 0.025). The proportion of Pro/Pro was higher in cervical carcinoma than that in normal control. The omni-constituent ratio was statistically different between HPV 16 positive and negative groups of cervical carcinoma in the Uigur group (chi(2) = 7.177, P < 0.05). The proportion of Arg/Arg was higher in HPV 16 positive group than that in HPV 16 negative group.

CONCLUSIONSp53 Arg72Pro polymorphism may be associated with the development of cervical carcinoma in Uigur and Han women in Xinjiang. p53 Arg/Arg genotype may be a genetically susceptible factor to HPV-associated cervical carcinoma in Uigur. p53 Pro/Pro genotype may be a genetically susceptible factor to cervical carcinoma in Han. There may be different susceptibilities to cervical cancer between Uigur and Han women in Xinjiang.

Adult ; Aged ; Alleles ; Base Sequence ; China ; ethnology ; Codon ; DNA, Neoplasm ; genetics ; Exons ; Female ; Gene Frequency ; Genes, p53 ; Genetic Predisposition to Disease ; ethnology ; Human papillomavirus 16 ; isolation & purification ; Humans ; Middle Aged ; Molecular Sequence Data ; Papillomavirus Infections ; ethnology ; Polymorphism, Genetic ; Tumor Suppressor Protein p53 ; genetics ; Uterine Cervical Neoplasms ; ethnology ; genetics ; virology ; Young Adult

OBJECTIVESTo study the correlation between positive rates of RNA in clinical confirmed severe acute respiratory syndrome (SARS) patients and its appearance in relation to the development of the disease in order to provide scientific basis for early diagnosis, effective prevention and treatment of the disease.

METHODSOne-step reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the SARS RNA in the clinical specimens from different courses of the disease. The representative amplicons were then sequenced. Chi-square for trend test was performed to study the correlation between positive rates of RT-PCR and at different periods after the onset of the disease.

RESULTSThe fragments amplified from the sputum specimens of SARS patients were shown to share 100% homology with the published SARS-associated coronavirus. Of the different clinical specimens, positive rate in the stools appeared to be the highest (21.55%). Chi-square for trend test revealed that the positive rates of stools and sputa of SARS patients decreased with the development of the disease (chi(2) for trend = 12.55 and 16.408, P = 0.0004 and P = 0.000 05 respectively).

CONCLUSIONOne-step RT-PCR proved to be an effective method for the detection of SARS-associated coronavirus from clinical specimens. Data as indicated that the positive rates of SARS coronavirus were decreasing in SARS patients along with the disease progression.

Adolescent ; Adult ; Aged ; Chi-Square Distribution ; China ; Disease Progression ; Feces ; virology ; Female ; Humans ; Male ; Middle Aged ; Mucus ; virology ; RNA, Viral ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; Severe Acute Respiratory Syndrome ; pathology ; virology