Next generation sequencing (NGS),also known as massively parallel sequencing (MPS),with significant advantages such as high throughput,rapidness,integration and low cost,has been widely used in research and clinical diagnosis fields and promises bright future for forensic applications.Compared with traditional DNA sequencing methods,NGS technologies allow the simultaneous detection of various forensic genetic markers.The classical CE-based STR genotyping method only can differentiate alleles with length polymorphism.However,there are occasions like DNA base substitution and repeat sequence variations,which give different alleles with the same size of amplicons.These alleles are heritable from parents to off-springs,and could be of great importance for solving criminal cases.NGS-based STR genotyping can be compatible with current STR data output mode,and allows for the full resolution of STR loci by providing additional sequencing polymorphism information.In this article,the authors focused on the forensic STR genetic markers and next generation sequencing,and reviewed recent progresses on NGS-based STR genotyping.Perspectives of NGS-based full resolution STR genotyping were presented and possible challenges were discussed in the hope of providing a reference for related studies and applications.

Objective To study genetic alterations in 9 STR loci and the Amelogenin locus in various tumor tissues. Methods twenty cancer tissues taken from 20 different unrelated individuals and their blood specimens were examined with Chelex-100 extraction of DNA, Profiler Plus PCR amplification and 310 Genetic Analyzer. Results All of the 10 STR loci exist genetic alterations. The genetic alterations occurred in 6out of 20 cases. The rate of genetic alteration was 30%. Six genetic alterations were found in one tumor tissue. Conclusion The forensic community has to take be cautious not to use the tumor tissue for personnel identification.

Objective To investigate the principles of diagnosis and treatment of postoperative intestinal obstruction.Methods The clinical datas of 89 cases of postoperative intestinal obstruction treated from 2001 to 2005 were analyzed retrospectively.Results All the 89 cases were cured without severe complications such as intestinal fistula and no death.The early postoperative inflammatory intestinal obstruction occurred in 2 cases.Recurrent intestinal obstruction occurred in 2 cases.Conclusions Intestinal obstruction can be caused by operation in abdomen.The diagnosis depends on the medical history,clinical manifestations,abdominal plain X-ray and computed tomography.Therapeutic procedure and operative opportunity play an important role in the treatment of postoperative intestinal obstruction.Non-operative approach should be the first choice for early postoperative inflammatory intestinal obstruction.Prompt surgery is clearly indicated and valuable for acute or being strangulative obstruction.Active operative treatment should be considered in recurrent chronic intestinal obstruction.

Objective To summarize the clinical experience of laparoseopie chdeeystectomy(LC).Methods Clinical records of 276 cases treated by LC were retrospectively reviewed.Results LC Was successfully accomplished in 265 cases and conversion to open surgery was required in 11 eases;The success rate of operation was 96%;and conversion rate of opening surgery was 4%;In the study,no severe complications occurred.Conclusion selecting fitable inpatients;With perfect technique and highly responsibility,LC could be performed safely.

OBJECTIVE: To analyze the clinical manifestations, diagnosis, therapy and prognosis of Langerhans cell histiocytosis (LCH) of the temporal bone in children. METHOD: Seven children with LCH of the temporal bone n our hospital were retrospectively summed up from April 2009 to April 2014. The patients were followed up 1-5 years, their clinical manifestations, imaging findings, diagnosis, therapy and prognosis were studied. Correlation between clinical classifications and prognosis was also analyzed. RESULT: Among the 7 patients, 4 were boys and 3 were girls. 5 cases belonged to the single system group and 2 cases belonged to the multisystem group. The most common clinical characters were temporal tumor, otorrhea, otalgia, hearing loss and granulation of external auditory canal. CT of the temporal bones showed extensive osteolytic destructions with diffuse soft tissure density, without border sclerotization. The cases were. received different therapies. Followed up for 1-5 years, 4 cases were regressive, 1 case kept stable, while 2 cases showed progressive. The two boys then received standard treatment combined steroids with vinblastine. The prognosis in the multisystem group was significantly different from the single system group (P < 0.05). CONCLUSION The clinical manifestations of LCH vary a lot. The diagnosis is based on histological and immunophenotypic examination of lesion tissue. The main therapy includs surgery, chemotherapy and radiotherapy. The prognosis of the single system group is much better than the multisystem group.
Child ; Combined Modality Therapy ; Deafness ; etiology ; Ear Canal ; pathology ; Ear Diseases ; etiology ; Female ; Histiocytosis, Langerhans-Cell ; diagnosis ; pathology ; therapy ; Humans ; Male ; Prognosis ; Retrospective Studies ; Steroids ; therapeutic use ; Temporal Bone ; pathology

Objective:To investigate the ethical cognition of human sperm among the volunteers who intend to donor sperm,and to analyze the voluntteers' thoughts for donor behavior and its countermeasures.Methods:A questionnaire survey was conducted to investigate the volunteers who were the first time consulted sperm donation in the human sperm bank of the affiliated reproductive hospital of Shandong university.The cognition on the purpose of sperm,the causes of sperm shortage,privacy protection of the offspring,and the future of sperm banks in China were collected.Results:Donors were not well known the uses of sperm and privacy protection of the offspring.They thought traditional values and ethical worry caused sperm shortage and worried about the future of human sperm banks in China.Conclusion:The overall cognition on the sperm bank of the volunteers still exist certain one-sidedness and blind,which has seriously affected the enthusiasm of volunteers to donate sperm.The relevant departments should pay more attention to this,focus on promoting the propaganda and supervision work of ethical system of human sperm bank,construct objective,justice,and dedicative atmosphere for sperm donation,in order to dispel the volunteers' psychological concerns,and promote the healthy,fast and sustainable development of human sperm bank.

ObjectiveTo explore the mechanism of extracorporeal shock wave (ESW) in treating osteogenic disorders and the ideal energy level. MethodsAfter success in marrow aspiration from patients' iliac crest, hMSCs were isolated by Percoll density gradient centrifugation and cultured in Dulbecco's modified Eagle's medium in a 5% CO2 and 37 ℃ incubator. Optimal ESW dose was determined by MTT of kinase-marked cytobiology. After hMSCs were exposed to ESW, their morphocytologic change, rate of adherence and doubling time were observed with IPCM. Enzyme cytochemistry reaction for the activity of alkaline phosphatase was also examined. ResultsESW of 5 kV and 100 times could increase cells' viability and proliferation (P<0.01), but higher than 7 kV would inhibit them. Rate of adherence of hMSCs in exposure group of passage 5 reached to 61.54%, which was significantly different from control group(P<0.05). Compared with control group, the MSCs' doubling time was short for 1.72 d (P<0.05). The curve of normal alkaline phosphatase activity of hMSCs was like type S, but ESW shortened its latent period, and promoted its peak time, which was significantly different from control group.ConclusionESW of 5 kV and 100 times can optimally promote the proliferation and activity of osteogen of hMSCs in vitro.

Objective To genotype mixed samples with next generation sequencing and evaluate its prospects in forensic DNA application. Methods Three mixed biological samples from rapes cases and their reference samples were collected. DNA was extracted using the MagAttract M48 DNA Manual Kit(200). The ForenSeqTMDNA Signature Prep Kit was used for library preparation, and next generation sequencing was performed on the MiSeq FGx system. The ForenSeqTMUniversal Analysis v1.2.1 software was used for data analysis. NGS-based STR results were compared with CE-based genotypes. Results A single length polymorphic STR allele in the mixed profile could be recognized as two sequence polymorphic STR alleles from different donors, which would assist mixed profile analysis. Such phenomenon was observed in D3S1358, D9S1122 and D13S317 in this work. Conclusion Our results suggested that precision STR genotyping of mixed samples based on NGS can provide more information and hints for mixed STR profile separation.

OBJECTIVE: To establish the kanamycin-induced deafness model in SD rats, and to investigate the expression and significance of transmembrane protease, serine 3 (TMPRSS3) in the cochlea following kanamycin ototoxicity. METHODS: A total of 40 male SD rats were randomly divided into 4 groups. The experimental rats received intramuscular kanamycin sulfate for 3, 7, and 14 consecutive days, and the control group were treated with normal saline for 14 days. Auditory brainstem responses (ABR) were obtained before and after the kanamycin administration. The expression of TMPRSS3 in the cochlea was identified and detected by immunohistochemistry and Western blot. RESULTS: Kanamycin-induced deafness model in the SD rats was successfully established. ABR thresholds were increased and the expression of TMPRSS3 in the cochlea was reduced after the kanamycin injection (P<0.01). CONCLUSION TMPRSS3 may play an important role in normal cochlea function and involve in the process of aminoglycoside antibiotics induced deafness.
Animals ; Anti-Bacterial Agents ; toxicity ; Cochlea ; drug effects ; metabolism ; Deafness ; chemically induced ; metabolism ; Disease Models, Animal ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Kanamycin ; toxicity ; Male ; Membrane Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serine Endopeptidases ; metabolism

OBJECTIVE: To observe the expression of proteinase transmembrane protease, serine 3 (TMPRSS3) in mouse cochlea, and to investigate the significance of TMPRSS3 in the inner ear. METHODS: The protein expression of TMPRSS3 in C57/BL mouse cochlea was identified and detected by immunohistochemistry and immunofluorescence. Different cochlear tissues, such as spiral ganglion neurons, corti organ, stria vascularis and so on, were separated to detect the gene expression of TMPRSS3 by real-time fluorescence quantitative polymerase chain reaction (qPCR). The cochlear tissues with different ages were collected and the expression of TMPRSS3 mRNA was detected by qPCR. RESULTS: TMPRSS3 was mainly expressed in the spiral ganglion neurons, and there was TMPRSS3 mRNA in the cochlea in groups with different age. The expression level of TMPRSS3 mRNA was much weaker. CONCLUSION The distribution of TMPRSS3 was observed in many regions of the mouse cochlea, but mainly in the spiral ganglion neurons. This indicates that TMPRSS3 may be involved in the physiological functional regulation of the spiral ganglion neurons.
Animals ; Cochlea ; metabolism ; Female ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; Mice, Inbred C57BL ; RNA, Messenger ; genetics ; metabolism ; Serine Proteases ; genetics ; metabolism