Ahigh-resolutionmethodforhumanleukocyteantigen-B(HLA-B)genotypingwasestablished based on the optimized polymerase chain reaction, cloning and sequencing technology. The exon2 and exon3 of HLA-B gene were amplified with primers based on the HLA-B gene sequence. These produced heterozygous alleles were effectively cloned into plasmid DNA based on the principle of plasmid incompatibility, and were followed by bacterial culture. Then Sanger sequencing was carried out and after analyzing the result by software ClustalX2 and IMTG/HLA database comparison, the HLA-B genotype of the samples was achieved. Seven clinical samples were detected, and the results were consistent with those of PCR-SBT genotyping method. The method was cost-effective, high-resolution and it did not require technical software. The use of universal primers simplified the cumbersome design and optimization process of specific primers.

Objective To study the method and clinical value of preservation of intercostobrachial nerve( ICBN) by fat dissolving meth-od during breast cancer operation. Methods The clinical data of 50 cases withⅠ~Ⅲa stage breast cancer from January 2013 to June 2013 were analyzed. Fifty patients were randomly divided into two groups,there were 26 patients in preservation group,whose ICBN were preserved by fat dissolving method during axillary lymph nodes dissection,and 24 patients in resection group,whose ICBN were not preserved by routine method during axillary lymph nodes dissection. Comparison of operation times,bleeding volume,the number of axillary lymph nodes dissection and upper arm sensory function of patients after operation between both groups was done. Results The mean time of operation was (102. 3 ± 15. 6) min in preservation group and(95. 6 ± 12. 4) min in resection group,while the number of axillary lymph nodes dissection was (19. 5 ± 8. 8 ) in preservation group and ( 19. 2 ± 9. 5 ) in resection group, with no significant difference between both groups (P>0. 05). Bleeding volume was (51. 2 ± 11. 5)mL in preservation group and (98. 5 ± 13. 4)mL in resection group,with significant differ-ences(P<0. 05). After postoperative one month,we observed upper arm sensory function of patients. It showed that 3 cases of sensory numb-ness or pain occurred in preservation group (11. 5%),20 cases of sensory abnormality occurred in resection group (83. 3%),mainly as sen-sory loss,numbness,pain or burning sensation,there was significant difference between both groups (P<0. 05). All patients were followed up half a year,patients with sensory abnormality in preservation group recovered,and recovery in resection group was not obvious,it still showed sensory abnormality in varying degrees. During the follow-up,no local recurrence or distant metastasis was found in both groups. Conclusion Preserving intercostobrachial nerve by fat dissolving method in breast cancer operation is based on conventional operation and made a few of improvements. It is simple and feasible. During the operation,we find that the axillary neurovascular is clearer,preservation of ICBN is easier. It does not affect the axillary lymph node dissection and operation time,while it can reduce incidence of postoperative sensory abnormality and improve the quality of life of patients,therefore it is worthy of clinical application.

Pyrosequencing is one of the important genetic polymorphism detection methods currently, but the complicated pretreatment procedure limits its application in clinical test. To simplify the whole process of pyrosequencing, on the basis of the linear_after_the_exponential_polymerase chain reaction ( LATE_PCR) , we improved the primer design method of LATE_PCR, increased the length and the concentration of the excess primer, applied direct amplification technology with whole blood, and established a whole blood_imLATE_PCR method based on common rTaq polymerase and “HpH Buffer” ( High pH buffer ) . The amplification system was optimized, and the influences of blood anticoagulant and the amount of whole blood template were investigated. The single stranded template for the pyrosequencing was obtained by PCR amplification using a single tube in one_step process, and the alcohol dehydrogenase gene polymorphisms of 24 clinical blood samples were then detected successfully. The results could be used to guide clinical individualized medication. The genotypes of ADH1B locus of 24 samples were 6 cases of AA homozygote, 14 cases of AG heterozygote, and 4 cases of GG homozygote. The genotypes of ADH1C were 20 cases of GG homozygote, 4 cases of AG heterozygote, and no cases of AA homozygote.

Objective To explore the clinical efficacy of biofeedback electrical stimulation plus pelvic floor muscle training (PFMT) in the treatment of stress urinary incontinence (SUI) after total hysterectomy.Methods The research objects who underwent total hysterectomy and complicated SUI were selected from January 2014 to December 2017 in our hospital.They were randomly divided into two groups:the study group (n =55) received biofeedback electrical stimulation plus PFMT treatment,while the control group (n=55) received PFMT treatment only.To record and compare the efficacy related indicators of the two groups.Results The indexes and scores of 3/6 months in the two groups were significantly improvedafter treatment.After 3 months the treatment,the study group had less leakage of urine [(4.3 ± 1.2)g vs (5.7 ± 1.9) g],better recovery of muscle strength of Ⅰ/Ⅱ muscle fibers [(3.5 ± 0.9) grade vs (2.8 ± 1.1) grade &(3.7 ± 0.6) grade vs (3.2 ± 1.2) grade],and better score of international consultation on incontinence questionnaire-short form (ICI-Q-SF) questionnaire [(2.7 ± 1.4) vs (4.5 ± 2.2)] compared with the control group,with statistically significant difference (P ≤ 0.05).After 6 months treatment,compared with the contiol group,the study group had less leakage of urine pad test [(1.4 ± 0.7) g vs (2.6 ± 1.1) g],better recovery of muscle strength of type Ⅰ/Ⅱ muscle fiber [(4.2 ±0.8) grade vs (3.5 ± 1.1)grade & (4.7 ±0.5) grade vs (4.0 ±0.9)grade],and better ICI-Q-SF score [(2.7 ± 1.4) vs (4.5 ±2.2)] (P ≤0.05).Conclusions Compared with using the PFMT alone,biofeedback electrical stimulation plus PFMT treatment had less leakage,better pelvic floor muscle strength and better subjective symptoms.

The article proposes the construction of network course of Chinese Medicine Preparation Analysis based on Black Board network teaching platform. It states the construction of teaching contents, the implementation of Case-Based Learning, the assistant of practical teaching and the formative evaluation. And it is helpful to improve the quality of classroom teaching, stimulate students self-regulated learning, expand their knowledge, improve their practical ability and innovative consciousness.

Objective To explore the expression of HR and Her?2 in breast cancer primary tumor and axillary lymph node metastasis. Methods Four hundred and twenty?eight female patients with unilateral breast cancer combined with axillary lymph node metastasis treated in the Affiliated Suqian Hospital of Xuzhou Medical University from January 2011 to January 2016 were selected in this study. Immunohistochemistry was used to detect the expression of ER,PR,Her?2 and Ki67 in primary tumor and axillary lymph node metastasis. Results The positive rates of ER expression were 75. 9% ( 325/428 ) and 70. 3% ( 301/428 ) respectively in primary tumor and axillary lymph node metastasis. The positive rates of PR expression were 61. 4% ( 263/428) and 56. 1% ( 240/428 ) respectively in primary tumor and axillary lymph node metastasis. The rates of Her?2 overexpression were 20. 1% ( 86/428) in primary tumor and the positive rate of Her?2 in axillary lymph node metastasis was 22. 7%( 97/428 ) . The positive rates of Ki67 expression were 45. 6%( 195/428 ) and 39. 7%(170/428) respectively in primary tumor and axillary lymph node metastasis. The expression of ER,PR,Her?2 and Ki67 in primary and axillary lymph node metastasis showed no statistical significance ( P>0. 05 ) . The molecular typing of primary tumor and axillary lymph node metastasis were not consistent in 31 patients ( 31/428,7. 24%) ,including 14 cases of primary tumor Luminal A,9 cases of Her?2 overexpression in axillary lymph node metastasis and 5 cases of triple negative breast cancer. Primary tumor Luminal B was detected in 10 cases, while 6 cases of Her?2 overexpression in axillary lymph node metastasis and 4 cases of triple negative breast cancer. Primary tumor Her?2 was overexpressed in 4 cases,while 1 case of Luminal A,3 cases of Luminal B in axillary lymph node metastasis. There were 3 cases of primary tumor triple negative breast cancer,while 2 cases of Luminal B in axillary lymph node metastasis and 1 case of Her?2 overexpression. Conclusion The expressions of ER, PR, Her?2 and Ki67 in primary tumor and axillary lymph node metastasis of some breast cancer were different. Immunohistochemistry for primary tumor and axillary lymph node metastasis of stage II?III breast cancer patients should be routinely carried out. Based on molecular typing of primary tumor and axillary lymph node metastasis,individualized treatment plan can be developed,so that patients will benefit from it.

The flipped class model is student-centered,which turns the traditional "teaching firstlearning behind" into the "learning first-teaching behind".The flipped class model can improve students' autonomous learning ability and other abilities.This article introduces the application of the flipped class in the course of medical immunology,including the course content selection,building QQ group learning platform,the production and release of independent learning resources,enactment of the task order and learning requirements,the design and implementation of classroom teaching activities and so on.The review and reflection of the formative assessment is also carried out.Through the reflection of the problems in the teaching,it is expected to lay the foundation for the promotion of the overturned class.

In order to explore the treatment of Her-2 positive breast cancer patients who failed in multi-line treatments, we retrospectively analyzed the clinical data of a patient with refractory Her-2 positive breast cancer.The patient was initially diagnosed as Her-2 positive advanced breast cancer.After six line treatment in the outer hospital, the patient′s condition was basically in a progressive state.The breast tumor was broken and purulent, the lung metastasis increased, and the patient′s quality of life was poor.The patient was admitted to Department of Breast Surgery of Affiliated Suqian Hospital of Xuzhou Medical University, after MDT discussion, we gave pyrrolotinib combined with capecitabine treatment, the chest wound healed gradually, the lung metastasis gradually reduced, and the quality of life was better.A retrospective analysis of this case showed that pyrrolidine combined with capecitabine may bring hope to Her-2 positive breast cancer patients who failed to receive multi-line therapies, especially those who failed to target therapy.

OBJECTIVETo analyze (CGG)n repeats sequence and AGG interspersion correlated with unstable expansion of FMR1 gene in a general Chinese population.

METHODSAmplideX FMR1 PCR Kit was used to amplify 380 X chromosomes from randomly selected 176 males and 102 females, 11 permutation carriers and 10 full mutation patients have served as controls. Results of capillary electrophoresis were analyzed with GeneMapper software Version 4.0. SPSS 11.0 software was used for statistical analysis.

RESULTSThe ratio of heterozygous females was 64.70%. The number of alleles in general males and females was 15 and 14, the classes of AGG pattern was 26 and 27, respectively. The range of alleles was between 17 to 45 CGG repeats in males and 21 to 44 CGG repeats in females, and 1 male was identified as gray zone carrier. The most frequent allele was 29 CGG repeats, which was followed by 30 and 36 repeats, while 28 CGG repeats were absent. The most common AGG pattern was 9A9A9, 99.21% of the population was detected with different forms and numbers of AGG interruption, and 6A interruption pattern was found in 10.02% samples especially in individuals with more CGG repeats. However, 57.58% of control samples had no AGG interruption, and none of the controls had 6A interruption pattern. No significant difference was observed in allele frequent distribution of (CGG)n repeats and AGG interspersion patterns between the males and females (P > 0.05), and AGGs was significantly different between general population and controls (P < 0.05).

CONCLUSIONAGGs and AGG pattern may have important roles in maintaining (CGG)n stability in general population of China, 9A9A6A9 may be a special pattern for preventing (CGG)n unstable expansion in Asian populations.

Adolescent ; Adult ; Alleles ; Female ; Fragile X Mental Retardation Protein ; genetics ; Fragile X Syndrome ; genetics ; Humans ; Male ; Middle Aged ; Trinucleotide Repeats

Objective To clarify the age patterns and types of differences so as to provide reference on prevention and interventions of hand,foot and mouth disease (HFMD) cases,in Hubei province.Methods We collected the HFMD case information of Hubei province from the Chinese National Notifiable Infectious Disease Reporting System in 2009-2015 while the information on pathogens from the laboratory monitoring system of Center for Disease Control and Prevention at all levels in Hubei province.All the data were stratified by age,disease severity,laboratory confirmation status,and serotypes of enterovirus.Results There were 495 783 reported HFMD cases from 2009 to 2015,in Hubei province,of which 1 045 were severe with 99 fatal.The annual notification rate was 1 231.0/106.HFMD cases were concentrated mainly in 0.5-5 year olds,with highest severity and mortality seen in 6-11 month-olds.The predominated pathogen in mild laboratory-confirmed cases each year,in order during 2009-2015 as:EV71,Cox A16,Cox A16,Cox A16,EV71,Cox A16 and other EV.HFMD showed semiannual peaks in April-June,November-December,and with more cases in the even years than in the odd years.Conclusions Children aged 0.5 to 5 years with 6 to 11 month-olds in particular,were the focused groups of attention in Hubei province.Our findings provided evidence for the improvement on monitoring program.Targeted intervention approaches should be strengthened to reduce the mortality and morbidity of HFMD in the province.