DNA genetic markers have always played important roles in individual identification, kinship analysis, ancestry inference and phenotype characterization in the field of forensic medicine. DNA methylation has unique advantages in biological age inference, body fluid identification and prediction of phenotypes. The majority of current studies independently examine DNA and DNA methylation markers using various workflows, and they use various analytical procedures to interpret the biological information these two markers present. Integrated methods detect DNA and DNA methylation markers simultaneously through a single experimental workflow using the same preparation of sample. Therefore, they can effectively reduce consumption of time and cost, streamline experimental procedures, and preserve valuable DNA samples taken from crime scenes. In this paper, the integrated detection approaches of DNA and DNA methylation markers on different detection platforms were reviewed. In order to convert methylation modifications to detectable forms, several options were available for pretreatment of genomic DNA, including digestion with methylation-sensitive restriction enzyme, affinity enrichment of methylated fragments, conversion of methylated or unmethylated cytosine. Multiplexed primers can be designed for DNA markers and converted DNA methylation markers for co-amplification. The schemes of using capillary electrophoresis platform for integrated detection add the pretreatment of genomic DNA on the basis of detecting DNA genetic markers. DNA and DNA methylation markers are then integrated by co-amplification. But the limited number of fluorescent options available and the length of amplicons restrict the type and quantity of markers that can be integrated into a panel. Pyrophosphate sequencing also supports integrated detection of DNA and DNA methylation markers. On this platform, due to the conversion of unmethylated cytosine to thymine after treatment with bisulfite, the methylation level of CpG site can be directly calculated using the peak height ratio of cytosine bases and thymine bases. Therefore, the methylation levels and SNP typing can be simultaneously obtained. However, due to the limited read length of sequencing, the detection of markers with longer amplicons is restricted. It is not conducive to fully interpret the complete information of the target sequence. Next-generation sequencing also supports integrated detection of DNA and DNA methylation markers. A preliminary experimental process including DNA extraction, pretreatment of genomic DNA, co-preparation of DNA and DNA methylation library and co-sequencing, has been formed based on the next-generation sequencing platform. It confirmed the feasibility of next-generation sequencing technology for integrated detection of DNA and DNA methylation markers. In field of biomedicine, various integrated detection schemes and corresponding data analysis approaches of DNA and DNA genetic markers developed based on the above detection process.Co-analysis can simultaneously obtain the genomic genetic and epigenetic information through a single analytic process. These schemes suggest that next-generation sequencing may be an effective method for achieving more accurate and highly integrated detection, helping to explore the potential for application in forensic biological samples. We finally explore the impact of interactions between sites and different pretreatment methods on the integrated detection of DNA and DNA methylation markers, and also propose the challenge of applying third-generation sequencing for integrated detection in forensic samples.

OBJECTIVE: To establish a secondary hemophagocytic lymphohistiocytosis(HLH) mouse model, and to investigate the effect of ruxolitinib on the disease manifestation of model mice. METHODS: Wild type C57BL/6 mice were randomly divided into 4 groups: two groups of mice were intraperitoneally injected with CpG oligodeoxynucleotide 1826 (CpG-ODN1826) every other day to induce HLH, and other two groups were control groups. One group of the CpG-ODN1826 groups and one of the control groups were given ruxolitinib, and other two groups were given the same amount of PBS. Blood samples, serum ferritin and hepatic/spleen weights of experimental mice were detected and serum cytokine levels were measured by ELISA. RESULTS: Compared with the control groups, the levels of white blood cells, hemoglobin and platelets in the CpG-ODN1826 groups were significantly lower (P＜0.05); and liver/body weight, spleen/body weight, serum ferritin, sCD25, IL-10, IL-1β, IFN-Ƴ, IL-12p70, GM-CSF, TNF-α and IL-18 levels significantly increased (P＜0.05). There was no significant difference in the levels of IL-2, IL-4, IL-5, IL-6, IL-22, IL-13, IL-27 and IL-23 between the two groups (P＞0.05). The spleen in CpG group had disordered internal structure, expanding red pulp and hyperplastic nucleated cells. The liver had severe perivascular inflammations. The spleen/weight of the ruxolitinib-treated mice in the CpG-ODN1826 group was significantly smaller than that of the unapplied ruxolitinib (P＜0.05). CONCLUSION The CpG-ODN1826 can induce secondary HLH symptoms in wild type C57BL/6 mice. Ruxolitinib can alleviate the symptoms of splenomegaly in HLH model mice.
Animals ; Disease Models, Animal ; Lymphohistiocytosis, Hemophagocytic ; Mice ; Mice, Inbred C57BL ; Pyrazoles

Objective The aim of the present study was to investigate the relationship between the intake of salt and salted food and the infection of Helicobacter pylori (Hp) among 40-69 years old local residents in a county with high gastric cancer risk in Anhui province. Methods From July 2015 to August 2018, we conducted a questionnaire and a serological test for Hp among 40-69 years old local residents in Lujiang county, Anhui province. The questionnaire focused on the consumptions of salt and salted food. The relationship between Hp infection and risk factors was analyzed by gender. Univariate and multivariate logistic regression analysis were used to analyze the relevant influencing factors. Results The Hp infection rate of total local residents was 50.07%. Among male subjects, age, body mass index(BMI), marital status, educational level, job, labor intensity and income had no link to Hp infection (all P>0.05). But among female subjects, BMI was associated with Hp infection ( 2=13.454,P=0.001). Besides, alcohol consumption was a risk factor for Hp infection in male subjects(OR=1.789，95% CI:1.188-2.694，P=0.003). But, high intake of salt and salted food had no effect on Hp infection after adjustment for alcohol consumption variable in men using multivariate analysis (all P>0.05). After adjusted for BMI variable among female individuals, high salt intake (≥9 g/day) (OR=1.462，95% CI:1.060-2.015，P=0.021) and the high salted food intake (≥1 times /day) were risk factors for Hp infection in women(OR=1.560，95% CI:1.021-2.383，P=0.040). Conclusions In one county with high gastric cancer risk in Anhui province, high salt intake (≥9 g/day) and high salted food intake (≥1 times/day) are risk factors for Hp infection among 40-69 years old female local residents.

OBJECTIVE: To investigate the relationship between spine-pelvic sagittal parameters and clinical efficacy before and after oblique lumbar interbody fusion(OLIF). METHODS: A retrospective analysis of clinical data of 65 patients with lumbar degenerative diseases treated with OLIF were performed from July 2017 to July 2018. There were 26 males and 39 females aged from 33 to 79 years old with an average of (62.72±10.23) years old. Oswestry Disability Index (ODI) and visual analogue scale (VAS) before and at the latest follow up were evaluated. Disc height (DH) and spine- pelvic sagittal parameters of the surgical segment were measured before and at the latest follow- up, including pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS) and lumbar lordosis (LL). According to the difference of PI-LL, it was judged whether PI and LL match and the patients were grouped, PI-LL ranged from -9° to 9° was set as matching group, and PI-LL less than -9° or larger than 9° was set as mismatching group. The spine-pelvic sagittal parameters were analyzed before and at the latest follow-up of OLIF in patients with lumbar degenerative diseases, and the correlation between changes and clinical efficacy was compared. RESULTS: All patients were followed up from 8 to 20 months with an average of (14.20±3.68) months. Operation time was (91.54±25.97) min, intraoperative blood loss was (48.15±10.14) ml, and the hospitalization time ranged from 6 to 19 days with an average of (9.28± 2.50) days. Totally 84 surgical levels, 46 patients were single segment and 19 patients were double segments. VAS and ODI score were improved from (4.88±0.99) point, (67.60±13.73) % preoperatively to (2.85±1.30) points, (30.57±6.48) % at the latest follow-up. There were significant differences in VAS and ODI scores between before and at the latest follow-up. The sagittal parameters of LL, PT, SS, PI, PI -LL and the surgical level DH were (42.80 ±16.35)° , (23.22 ±10.91)° , (26.95 ± 13.30)°, (50.22±14.51)°, (7.53±16.13) °, (0.91±0.29) cm preoperatively and improved to the latest follow-up (49.95± 12.82) °, (17.94±9.24) °, (33.71±12.66) °, (51.65±10.26) °, (1.68±17.00) °, (1.20±0.40) cm;there were statistical differences in LL, PT, SS, PI-LL, DH before operation and at the latest follow up, while no difference in PI. LL of preoperative PI-LL in matched group was (48.76±11.09)° , and (38.00±18.37)° in PI-LL mismatch group, there was difference between two groups. There were no differences in VAS, ODI, PT, SS, PI and DH between two groups. At the latest follow-up, ODI between PI-LL matched group and PI-LL mismatched group were (29.40±5.93)% and (32.86±7.02)% respectively, and had difference in ODI between two groups;while there were no significant differences in VAS, LL, PT, SS, PI, and DH. Pearson correlation analysis showed preoperative PT-LL was positively correlated with VAS;PT was positively correlated with ODI at the latest follow-up. CONCLUSION OLIF has a good surgical effect on lumbar degenerative diseases, and could change spine-pelvic sagittal parameters of patient to a certain extent, and further restoring the balance of the sagittal plane of lumbar spine.
Adult ; Aged ; Female ; Humans ; Lumbar Vertebrae ; Lumbosacral Region ; Male ; Middle Aged ; Pelvis ; Retrospective Studies ; Spinal Fusion ; Treatment Outcome

Simiao Yong'an Decoction is composed of Lonicerae Japonicae Flos, Scrophulariae Radix, Angelicae Sinensis Radix, Glycyrrhizae Radix et Rhizome, which was chosen as one of the 100 classic prescriptions in Catalogue of Ancient Classics Prescription(the first batch). Through tracing to the source, It was found that the Simiao Yong'an Decoction(but not named) originated from the Shi Shi Mi Lu, and was later cited by books such as Ancient and Modern Book Integration-Full Record of Medical Department and New Edition of Useful Prescriptions. Literature shows that this prescription was not named until first reported in the Effect of Traditional Chinese Medicine on Arterial Embolism Gangrene in 1956 by a journalist LYU Min. This article recorded that SHIJIA Baoshan, a monk from Hebei Province, used self-named "Simiao Yong'an Decoction" to treat local arterial embolic gangrene. After comparison, there was two difference between ancient books and SHIJIA Baoshan's records. Firstly, according to ancient books, the composition and dosage of Simiao Yong'an Decoction is Lonicerae Japonicae Flos 90 g, Scrophulariae Radix 90 g, Angelicae Sinensis Radix 60 g, Glycyrrhizae Radix et Rhizome 30 g", and the ratio is 3∶3∶2∶1. By SHIJIA Baoshan's record, the composition and dosage are: Lonicerae Japonicae Flos 66 g, Scrophulariae Radix 132 g, Angelicae Sinensis Radix 99 g, Glycyrrhizae Radix et Rhizome 33 g, and the ratio changed to 2∶4∶3∶1. Secondly, ancient books show that patients can be healed after taking seven or ten days of the previous prescription, however, it would take 3 or 4 months, even 7 months in SHIJIA Baoshan's records. It can be considered that the previous prescription should be used at the beginning of gangrene, while the modified Simiao Yong'an Decoction by SHIJIA Baoshan is widely used in the middle and late stages of gangrene, even the critical condition, that is the reason for longer treatment and larger dosage. Nowadays, Simiao Yong'an Decoction is not limited to the treatment of gangrene and bulla in clinic. Relevant studies have confirmed that Simiao Yong'an Decoction has the effects such as anti-inflammatory, plaque stabilization, lipid-lowering, vascular protection, improvement of hemorheology, anticoagulation, inhibition of thrombosis and fibrinolysis, etc. In the follow-up, we should carry out the analysis of the compatibility of this four medicines, and redefine the scope of its clinical application under the theory of traditional Chinese medicine.
Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Plant Roots ; Rhizome

BACKGROUND: Important extracellular matrixes are reduced with the prolongation of duration of cyclic pressure in the endplate of the intervertebral disc. Meanwhile, the expression of Wnt-5a gene is significantly decreased. There is an important relationship between Wnt-5a gene and intervertebral disc degeneration (IDD). OBJECTIVE: To investigate the expression of Wnt-5a gene under cyclic pressure in a rabbit model of IDD and to explore its role in IDD progress. METHODS: Lumbar intervertebral discs were removed from the 6-month-old New Zealand white rabbits to prepare IDD models and were then randomly divided into experimental (cyclic pressure ) and control (no intervention) groups. The morphological changes of intervertebral discs were observed by hematoxylin-eosin staining and safranin O-fast green staining. The mRNA expression levels of proteoglycan, collagen type Ⅱ, and Wnt-5a were detected by real-time PCR. The protein expression level of Wnt-5a was detected by western blot assay. RESULTS AND CONCLUSION: The morphology of intervertebral discs cultured for 7 days in the experimental and control groups showed a certain change, but was still intact; expression levels of aggrecan, type Ⅱ collagen, Wnt-5a showed differences from the intervertebral discs cultured for 0 day. On day 14, the damage to the histomorphology was severer in the experimental group than the 0-day control group. The mRNA expression levels of proteoglycan, collagen type Ⅱ, and Wnt-5a were decreased in both groups, especially the experimental group, at 7 and14 days. The mRNA and protein expression levels of Wnt-5a revealed the same change trend with time. To conclude, regulation of Wnt-5a expression may alter the process of endplate cartilage degeneration, and thus providing new ideas for the prevention and treatment of IDD.

Objective To evaluate the clinical efficacy and safety of dimemorfan phosphate in the treatment of dry cough.Methods Three hundred seventy -eight patients with dry cough were randomly divided into experimental group ( n=216 ) , positive control ( n=108 ) and place-bo group ( n=54 ).Experimental group was taken dimemorfan phosphate 20 mg, orally, tid, 5 -7 d.Positive control group was taken dextro-methorphan 30 mg, orally, tid, 5 -7 d.Placebo group was taken dimemorfan phosphate simulation tablets 20 mg or dextromethorphan simulation tablets 30 mg, tid, 5-7 d.Comparison of the clinical effica-cy, change of the total cough score after 5-7 d treatment, and the inci-dence of adverse drug reactions between the three groups.Results The clinical effectiveness of cough symptom after treatment , experimental group and positive control group and placebo group were 81.02%, 84.26%and 38.89%, respectively.The difference between experimen-tal group with placebo group was statistically significant ( P <0.01 ) , there was no statistically significant difference between experimental group with positive control group ( P>0.05 ).The average decrease of total cough score after treatment in the experimental group , the positive control group and the placebo group were (2.34 ±1.42), (2.43 ±1.55) and (1.39 ±1.20), respectively.The decrease in the total cough score of experimen-tal group was superior to that of placebo group ( P<0.01 ) , the decrease in the total cough score of experimental group was not inferior to positive control group ( P>0.05 ).The incidence of adverse events were 4.21% in experimental group , 12.04%in positive control group and 5.66%in placebo group , respectively.Among these , the incidence of adverse events associated with drug were 1.87%, 10.19%, 5.66%, respectively.Conclusion Dimemorfan phos-phate has exact antitussive efficacy in the treatment of dry cough , has the same antitussive efficacy and similar side effect in the treatment of dry cough compared with the dextromethorphan.

OBJECTIVETo study the effectiveness of waist circumference cut-off values in predicting the prevalence of metabolic syndrome (MetS) and risk factors in adults in China.

METHODSA cross-sectional survey was condcuted in 14 provinces (autonomous region, municipality) in China. A total of 47,325 adults aged⋝20 years were selected by multistage stratified sampling, and questionnaire survey and physical and clinical examination were conducted among them. MetS was defined according to the International Diabetes Federation (IDF) criteria and modified IDF criteria.

RESULTSThe age-standardized prevalence of MetS was 24.2% (22.1% in men and 25.8% in women) and 19.5% (22.1% in men and 18.0% in women) according to the IDF criteria and modified IDF criteria respectively. The age-standardized prevalence of pre-MetS was 8.1% (8.6% in men and 7.8% in women) according to the modified IDF criteria. The prevalence of MetS was higher in urban residents than rural residents and in northern China residents than in southern China residents. The prevalence of central obesity was about 30% in both men and women according to the ethnicity-specific cut-off values of waist circumference for central obesity (90 cm for men and 85 cm for women). Multivariate regression analysis revealed no significant difference in risk factors between the two MetS definitions.

CONCLUSIONUsing both the modified IDF criteria and ethnicity-specific cut-off values of waist circumference can provide more useful information about the prevalence of MetS in China. Conclusion Using both the modified IDF criteria and ethnicity-specific cut-off values of waist circumference can provide more useful information about the prevalence of MetS in China.

Adult ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Male ; Metabolic Syndrome ; diagnosis ; epidemiology ; Middle Aged ; Obesity ; epidemiology ; Prevalence ; Risk Assessment ; Risk Factors ; Waist Circumference

BACKGROUNDChromosomal abnormalities have been shown to play an important prognostic role in multiple myeloma (MM). Interphase fluorescence in situ hybridization (i-FISH) has been much more effective to identify cytogenetic aberrations in MM than conventional cytogenetic technique (CC). To clearly determine the cytogenetic features of Chinese MM patients and identify their prognostic implications, we designed a multicenter study based on i-FISH including 672 patients from 52 hospitals in China.

METHODSAll 672 patients were systematically screened for the following genomic aberrations: del(13q), IgH rearrangement, del(p53) and 1q21 amplifications.

RESULTSThe analysis showed that the chromosomal changes were detected in 22.1% patients by CC and in 82.3% patients by i-FISH. The most common abnormalities by CC were chromosome 1 aberrations (48.4%), -13/13q- (37.6%), hyperdiploidy (36.6%), hypodiploidy (30.1%) and IgH rearrangements (23.7%). The most frequent abnormalities by FISH was del(13q), which was found in 60.4% patients, whereas IgH rearrangement, 1q21 amplification and p53 deletions were detected in 57.6%, 49.0% and 34.7% cases, respectively. By statistical analysis, -13/13q- by CC was associated with low level of platelet (P = 0.015), hyperdiploidy was associated with low level of serum albumin (P = 0.028), and IgH rearrangement by FISH was associated with high level of β2 microglobulin (P = 0.019). Moreover, 1q21 amplification and del(p53) by FISH conferred a high incidence of progressive disease (PD) after initial therapy. Metaphase detection of IgH rearrangements and chromosome 1 aberrations concurrently was associated with a short progression free survival (PFS) (P = 0.036). No significant prognostic implications of other cytogenetic abnormalities were found associated with overall survival and PFS.

CONCLUSIONSChinese MM patients had similar cytogenetic abnormalities compared with the previous reported studies. However, the prognostic significance of FISH aberrations were not clearly determined and further study is required.

Adult ; China ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology

Objective To explore whether the cellular apoptosis susceptibility (CAS) protein could serve as a pathologic marker for HCC diagnosis and the roles of CAS expression in HBV infection associated HCC.Methods The expression of CAS protein in HCC and its paracarcinoma tissues,non-tumor liver cirrhosis and hepatitis tissues were detected by immunohistochemistry.Meanwhile,HBsAg,HBcAg and HBV DNA in HCC tissues with HBV infection were examined by immunohistochemistry and in situ hybridization respectively.Results The expression of CAS protein was significantly higher in HCC than in its paracarcinomas tissues ( P ＜ 0.01 ),a.nd higher in paracarcinomas tissues than in non-tumor liver cirrhosis and hepatitis tissues ( P ＜ 0.01 ).Poorly differentiated tumors immunochemically stained stronger than moderately or well differentiated( P ＜0.01 ).CAS protein expression was significantly higher in HBV-infected HCC tissues than that of in non-HBV infection ( P ＜ 0.01 ).Meanwhile,in HBV-infected HCC tissues,the staining intensity score of CAS protein in HBV DNA positive HCC tissues was significantly higher than HBV DNA negative tissues ( P ＜ 0.05 ).Conclusions Higher expression of CAS protein is found in HCC tissues,and the intensity of CAS protein expression is related closely to tumor differentiation.We suggested that CAS protein might serve as a marker for HCC diagnosis and differentiation estimation,and deduced that CAS might play an important role in the initiation of HBV infection associated HCC through upregulating expression of CAS.