Objective To master of the characteristics of development and variation in plague natural foci of Marmota sibirica, and to analyze the plague monitoring results from 2004 to 2008. Methods From May to September every year, we monitored plague in Manzhouli, Chenbaerhuqi, Xinbaerhuzuoqi, Xinbaerhuyouqi,Ewenkeqi and Yakeshi city. The monitored area was 20 000 - 40 000 hm2 in every county. The density of Marmota,ground squirrel and moonlighting rats was surveyed by path method, one-day bow-clip method and a clamp was placed every five meters, respectively. The classification and identification of ectoparasite fleas were done by using low power lens. The fleas in the ground squirrel hole were collected by flannel rubber stick, then classified and identified. According to National Standard of Plague Diagnostic Criteria(GB 15991-1995), the collected rats and fleas were detected by isolation and cultured Yersinia pestis, the serums of collected rats were tested by indirect hemagglutination test. Results In five years, the density of Marmota and ground squirrel was 0.010/hm2 and 0.602/hm2, respectively. The capture rate of moonlighting rats was 2.69％ (258/9600). The flea infection rate of Marmota was 17.54％(10/57) and the fleas index was 2.54. The fleas infection rate of ground squirrel was 28.40％(213/750) and the fleas index was 1.01. Flea infection rat of ground squirrel hole was 5.60％(46/822) and the fleas index was 0.17. The total number of various hosts of Yersinia pestis detected was 1351, the groups number of the variety of cultured fleas was 127, and the pathogen test results were negative. The number of serum tested was 1064, positive number was 43, and the detection rate was 4.04％(43/1064). The highest positive titer was 1 : 1280. Other than 2006, the remaining four years were found positive for blood clotting material; positive serum was found in a total of three regions, they were Manzhouli, Xinbaerhuzuoqi, and Xinbaerhuyouqi; and 30, 12 and 1 copies of hemagglutination-positive sera were detected. Conclusions The epidemic of plague natural foci of Marmota sibirica is in a active state, and gradually expands the scope. We must continue to strengthen the inspection of the bacteria, bearing in mind the replacement of the region's main host, make every effort to prevent and control of human plague.

Objective To evaluate the clinical effect of the simple syringomyelia patients with syringopleural shunt (SPS) and syringosubarachnoid shunt (SSS).Methods Twenty-eight patients with syringomyelia were selected as our subjects.Of which,18 patients were operated with SPS and 10 cases were with SSS.The therapy effect was compared between groups.Results All patients were checked with MRI 3 months after the operations and showed that syrinx cavity was significantly narrow of all patients.At early stage (48 h) after surgery 9 cases in SPS and 4 cases in SSS were showed the decrease syrinx cavity.Among patients with SPS,15 cases (83.3％) had the symptoms and signs improved,1 case (5.5％) withno changes,1 case (5.5％) with worse effect,and 1 cases(5.5％) occurred the tethered spinal cord.Meanwhile,among,patients with SSS,8 cases(80.0％) had the symptoms and signs improved,1 case(10.0％) with no changes,and 1 case (10.0％) with worse effect.There were 4 patients with pneumothorax in SPS group after operations,and the lung compression ratios were less than 5％.These cases were not taken any special treatment for the pneumothorax.All patients in two groups had not been infected and pleural effusions.No cases had taken the tube plugging.Conclusion The simple syringomyelia patients with the spine injury should take the cavity shunt.The SPS has some advantages because it can provide the partly negative pressure.But it should be certified by more cases and a long-time follow up.

Abstract] Objective To establish an assay for the detection of porcine parvovirus ( PPV) and to verify its application for monitoring cells used for production.Methods A pair of primers and one probe were designed according to the conserved sequence encoding non-structural protein 1 (NS1).Based on the designed primers, a real-time fluorescent quantitative PCR assay for the detection of PPV was developed. Several parameters including the linearity, precision, minimum detection limit and anti-interference of the established assay were evaluated.A stock of PPV strains was prepared by infecting swine testicle ( ST) cells with PPV strains.An assay for the detection of PPV infection was developed by using ST cells as sensitive cells.A combined ST cell infection-PCR test was developed by combining the ST cell infection assay with the real-time fluorescent quantitative PCR assay.The sensitivity of ST cell infection-PCR test was analyzed.The cell samples used for production of biological products were detected by using the established assay.Results The real-time fluorescent quantitative PCR assay was specific for the detection of PPV without cross-reaction to other species of parvovirus virus, SV40 virus and other porcine viruses.The linear range of the assay was 1×109-1×104 copies/μl with a R2 value more than 0.98.The sensitivity of the real-time quantitative PCR assay was 1×104 copies/μl.Both of the intra-and inter-coefficient of variation (CV) were less than 5%in Ct values.The intra-and inter-CV in copies of detection were 5%-15% and 30%-40% respectively.The minimum detection limit of the real-time fluorescent quantitative PCR assay was 1CCID50/ml.The PPV strains were detected in cell samples with no interference.The sensitivity of ST cells infection-PCR test was 0.01CCID50/ml.All of the 22 cell samples were negative for PPV by using the real-time fluorescent quanti-tative PCR assay.Conclusion The real-time fluorescent quantitative PCR and the ST cell infection-PCR test for the detection of PPV in cells were established successfully.The application of the two assays was conducive to further enhance the safety of using cells for production and therapy.

Objective:To provide the basic data for the further revision of mycoplasma test method described in Chinese Pharma-copoeia and some references for the operation standardization of drug mycoplasma test. Methods:Two incubation conditions,namely aerobic conditions and microaerophilic conditions,were compared with respect to the growth status of mycoplasma in liquid and solid media. Results:The growth of mycoplasma was obvious difference between the two incubation conditions,and the microaerophilic con-ditions were better than the aerobic conditions. Conclusion:The microaerophilic conditions can be used in the incubation of mycoplas-ma test,which should be defined and standardized in the future Chinese Pharmacopoeia.

Objective:To investigate the growth-promoting properties and applicability of the mycoplasma test culture media pre-scribed in Chinese pharmacopoeia, and provide reference for the standardization of the drug mycoplasma test method. Methods: Re-spectively using four quantitative detection methods including sensitivity, degree of color change, colony counts and colony diameter, and mycoplasma media widely used in the world as the reference media, the growth-promoting properties of 4 batches of mycoplasma broth and 4 batches of arginine mycoplasma broth from four domestic manufacturers were investigated. Results:The results of sensitivity assay and absorbance detection showed that all the media inoculated with below 100 CFU test microorganisms exhibited visible color change. Furthermore, the results of color change degree and colony diameter showed that there were significant differences among the media products from different manufacturers(P<0. 01). Conclusion:Mycoplasma broth and arginine mycoplasma broth both can sup-port the growth of below 100 CFU test microorganisms. Due to the difference in the growth-promoting properties among the media prod-ucts from different manufacturers, the drug mycoplasma test workers should use more sensitive methods to examine the applicability of the media.

Objective To observe the changes of esophageal motility in patients with achalasia (AC) before and after peroral endoscopic myotomy (POEM ) and to evaluate the effects of POEM on esophageal motility in AC .Methods A total of 35 patients with AC received POEM .The esophageal motility in response to different food swallows (5 mL liquid and 2 cm × 2 cm × 2 cm solid food) was evaluated by high-resolution manometry (HRM) system before operation and one month after operation .The changes of parameters of esophageal body and lower esophageal sphincter (LES) were analyzed and compared before and after operation .The t-test ,non-parametric test or single factor analysis of variance was performed for statistical analysis .Results Before POEM operation ,lower esophageal sphincter pressure (LESP) of 35 patients in response to liquid swallows and solid swallows was (28 .94 ± 18 .70) mmHg (1 mmHg=0 .133 kPa) and (26 .41 ± 11 .57) mmHg ,respectively ;after operation it was (16 .02 ± 5 .46) mmHg and (15 .82 ± 5 .04) mmHg ,respectively ;and the differences were statistically significant (t= 4 .338 and 4 .726 ,both P<0 .01) .Before operation ,4 s integrated relax pressure (4 s IRP) during liquid swallows and solid swallows was (27 .18 ± 14 .63) mmHg and (28 .46 ± 11 .15) mmHg ,respectively ;after operation it was (12 .22 ± 6 .75) mmHg and (14 .54 ± 7 .83) mmHg ,respectively ;and the differences were statistically significant (t= 5 .902 and 5 .436 ,both P< 0 .01) .And after operation intra boluspressure (IBP) of liquid swallows and solid swallows also decreased compared to that before operation (t=5 .075 and 2 .944 ,both P< 0 .01) .Lower esophageal sphincter relaxation rate (LESRR) during liquid swallows and solid swallows increased compared to that before operation (t= -2 .990 , P< 0 .01 ;t=-0 .340 ,P>0 .05) .There was no difference in the distal contractile integral (DCI) and distal esophageal peristaltic amplitude in subtype Ⅰ and Ⅲ patients before and after operation (all P>0 .05 ) .All those parameters decreased in subtype Ⅱ patients (Z= -2 .704 and -2 .489 ,P< 0 .05 ;Z= -1 .929 and-0 .747 , P> 0 .05 ) . Proximal esophageal peristalsis was observed in two patients after operation , however there was no integrated esophageal body peristalsis .Clinical symptoms quickly relieved in all patients after POEM operation and clinical score significantly decreased compared to that before operation (0 .86 ± 1 .19 vs 8 .16 ± 1 .84 ,t=20 .605 , P<0 .05) .Conclusions POEM can effectively relieve LES relaxation disorder in AC patients and improve esophageal body peristalsis to a certain degree .The efficacy is regardless of AC types ,and further studies are need to shed light on the long-term efficacy .The long-term efficacy still need further follow-up study .

Objective To establish an assay for the detection of Torque teno sus virus ( TTSuV) strains and to analyze its preliminary application to biologics.Methods Primers and probe were designed according to the conserved sequences.A fluorescent PCR assay for the detection of TTSuV strains was estab-lished.Several parameters including the specificity, linearity, accuracy, sensitivity and anti-interference of the established assay were verified.The fluorescent PCR assay was performed to detect the samples of por-cine blood, cell substrate and rotavirus vaccine.The porcine blood samples positive for TTSuV strains were further genotyped.Results The established fluorescent PCR assay was confirmed to have high specificity as no cross-reactions with parvovirus virus of various species, SV40 and porcine circovirus strains were detec-ted.The linear range of the assay was 1×109-1×103 copies/μl for TTSuV1 genotype and 1×109-1×102 cop-ies/μl for TTSuV2 genotype with a R2 value more than 0.993.The sensitivity of the fluorescent PCR assay was 1×103 copies/μl for TTSuV1 genotype and 1×102 copies/μl for TTSuV2 genotype.The intra-and inter-CVs were both less than 7%in Ct values and less than 25% and 45% respectively in copies.No interfer-ence was found in the detection of TTSuV nucleic acids from cell samples.8 out of 20 porcine blood samples were positive for TTSuV strains, among which one sample was positive for TTSuV1 genotype, four samples were positive for TTSuV2 genotype and the rest were positive for both TTSuV1 and TTSuV2 genotypes.Com-pared with the reference strain, strains genotyped as TTSuV1 and TTSuV2 were respectively shared 98%-99%and 98%homologies in sequences.All of the cell substrate and rotavirus vaccine samples detected by the fluorescent PCR assay were negative for TTSuV strains.Conclusion The fluorescent PCR assay for the detection of TTSuV was established successfully, the application of which would further improve the safety of biologics.

AIM:To detect the expression of basic fibroblast growth factor (bFGF) and matrix metalloprotei-nase 9 (MMP9) in nasopharyngeal carcinoma (NPC) and its correlation with clinicopathological features and prognosis of the patients.METHODS:The expression of bFGF and MMP9 was detected by the method of SP immunohistochemical staining in biopsy tissues of NPC patients.The relationship between the expression and the clinical significance was analyzed as well.RESULTS:In 289 cases of NPC patients, the positive rates of bFGF and MMP9 were 71.3% and 61.6%, respectively.Correlation analysis demonstrated that the expression rates of bFGF and MMP9 were both positively associated with N stage and clinical stage in NPC patients.The high expression rates of both bFGF and MMP9 were associated with poor overall survival and progression-free survival of NPC patients.Furthermore, the positive rate of bFGF was positively correlated with that of MMP9, and over-expression of both bFGF and MMP9 was correlated with the poorest survival outcomes in NPC patients.CONCLUSION:bFGF and MMP9 are over-expressed in NPC tissues and significantly associated with NPC recurrence and poor outcome.The combined interpretation of bFGF and MMP9 expression levels leads to refinement of the risks for the NPC patients and could be chosen as the prognostic biomarkers.

In order to study genetic variation diversity of porcine circovirus type 2 (PCV2) strains in Shanxi,the genomic sequences of nine PCV2 strains including SXQX,SXCZ,SXTY2,SXJC,SXJX,SXLL,SXPY,SXPG and SXXY recently isolated from some areas of Shanxi from 2013 to 2016,was cloned,sequenced and received by GenBank.The amplified PCV2 genomic sequences,ORF2 sequences and Cap protein amino acid of these nine strains were analysed and compared with those of published 28 PCV2 strains by DNAStar,drawing phylogenetic tree.The results showed that the genomic sequences of SXJX,SXJC and SXXY PCV2 strains were 1 768 bp,and the others were 1 767 bp,which accounted for 33％ and 67％,respectively.The homologies of nucleotide sequences of the nine strains were 94.7％-99.8％,the homologies of nucleotide sequences of the nine strains with the 28 isolates from different regions of the world PCV strain were 93.9％-99.9％,and the homologies of nucleotide sequences of the nine strains with the domestic vaccine strains were 95.1％-99.8％.The phylogenetic analysed that SXJX,SXJC and SXXY belonged to genotype PCV-2D,SXLL,SXPY and SXCZ belonged to genotype PCV-1C,and SXTY14,SXPG and SXQX belonged to genotype PCV-1A/1B.Thus it proved that the epidemic strain of PCV2 was mainly PCV-2b in Shanxi.The homologies of ORF2 nucleotide sequences and Cap amino acid of the nine strains were 90.0％-100.0％ and 87.1 ％-100.0％ respectively,the homologies of ORF2 nucleotide sequences and Cap amino acid of the nine strains with the 28 isolates from different regions of the world PCV strain were 87.6％-100.0％ and 84.1％-100.0％ respectively,and the homologies of ORF2 nucleotide sequences and Cap amino acid of the nine strains with the domestic vaccine strains were 91.0％-100.0％ and 89.3％-100.0％ respectively.The Cap amino acids of SXQX,SXJX,SXTY14,SXPG,SXJC and SXXY PCV2 were 233,ORF2 of SXQX,SXTY14 and SXPG located at 1 033-1 734 bp,ORF2 of SXXY,SXJX and SXJC located at 1 033-1 734 bp,and the Cap amino acids of SXCZ,SXLL and SXPY PCV2 were 234,ORF2 of them located at 1 030-1 734 bp,in addition,the positions of 1 030-1 734 bp were more three bases TCA than other ORF2 genome sequence of 1 767 bp,resulting in increasing a K (Lys) of amino acid sequencein at the 234 position.Also Cap protein of 9 PCV2 strains showed more amino acid variation in addition to the only high-ly conserved glycosylation sites (NYS) (pp.143-145 amino acid).It provided theoretical basis for the PCV2 immune prevention of research in Shanxi,and the data of basic theory of molecular pathogenesis of PCV2.

OBJECTIVETo observe the efficacy and safety of Qizhi Jiangtang Capsule (QJC) in treating stage 3b diabetic kidney disease (DKD) patients with macroalbuminuria.

METHODSPatients who conformed to the diagnostic criteria of stage 3b DKD were randomly assigned to two groups according to random digital table, the experiment group and the control group, 84 in each group. All patients received a two-week elution period, and then were treated with basic Western therapy. Patients in the experiment group took QJC, 5 pills per time, 3 times a day, while those in the control group took Valsartan Capsule 160 mg each time, once daily. The observation period of follow-ups was limited within 6 months, and the time points were set as the baseline, 1st month, 3rd month, and 6th month. Systolic blood pressure (SBP), diastolic blood pressure (DBS), 24 h urine protein quantitative (24 h UPQ), plasma albumin (ALB), and serum creatinine (SCr) were detected and recorded, and estimated glomerular filtration rate (eGFR) was calculated. The occurrence of hypoglycemic reaction, coagulation disorder, gastrointestinal tract reaction, allergy, hyperkalemia, doubling of creatinine, and overall adverse events were observed and recorded at same time.

RESULTSFinally 81 patients in the experiment group and 80 patients in the control group were effectively included. Compared with the baseline level, SBP and DBS obviously decreased in the control group at month 1 of treatment (P < 0.05), and more significantly decreased at month 6 of treatment (P < 0.01). SBP at month 1, 3, and 6 of follow-ups; DBS at month 6 of follow-ups was lower in the control group than in the experiment group (P < 0.05). At month 1, 3, and 6 of follow-ups, 24 h UPQ of the experiment group was significantly lower than the baseline level (P < 0.01). It was also significantly lower than the level of the control group at the same time point (P < 0.05). There was no significant difference in 24 h UPQ at month 1, 3, and 6 of follow-ups between the control group and the baseline level (P > 0.05). ALB of the experiment group showed an increasing trend. It was significantly higher than the baseline level at month 6 (P < 0.05), which was also higher than that of the control group at same period (P < 0.05). There was no significant difference in the ALB level in the control group (P > 0.05). SCr of two groups showed an increasing trend. SCr of the experiment group was significantly higher at month 1, 3, and 6 follow-ups than the baseline level (P < 0.05). But the increment of SCr was higher in the control group than in the experimental group, and obviously higher than the baseline levels (P < 0.05). eGFR of both groups showed a decreasing trend. The decrement was higher in the control group than in the experimental group (P < 0.05). The proportion of progression of renal functions at month 1, 3, and 6 of follow-ups in the experimental group was 0.0% (0 case), 9.55% (8 cases), and 21.4% (18 cases), while they were 8.3% (7 cases), 21.4% (18 cases), and 40.5% (34 cases) in the control group. There was no statistical difference in the proportion of progression of renal functions between the two groups at month 3 and 6 of follow-ups (P < 0.05). There was no statistical difference in the incidence of adverse reactions between two groups (P > 0.05).

CONCLUSIONQJC could effectively reduce urinary protein of patients with stage 3b DKD, and delay the progression of renal functions.

Adult ; Albumins ; analysis ; Albuminuria ; drug therapy ; Blood Pressure ; drug effects ; Creatinine ; blood ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerular Filtration Rate ; Humans ; Male ; Middle Aged ; Tetrazoles ; therapeutic use ; Treatment Outcome ; Valine ; analogs & derivatives ; therapeutic use ; Valsartan