Objective:To analyze the clinical characteristics and genetic basis of a male patient with primary infertility caused by Acephalic spermatozoa syndrome.Methods:A patient who had presented at the Henan Provincial People′s Hospital on October 1, 2022 was selected as the study subject. Clinical data and results of laboratory exams and sperm electron microscopy were collected. The patient was subjected to whole exome sequencing (WES), and candidate variants were verified by Sanger sequencing and pathogenicity analysis.Results:WES revealed that the patient has harbored compound heterozygous variants of the PMFBP1 gene, namely c. 853del (p.Ala285Leufs*24) and c. 1276A>T (p.Lys426X), which were both unreported previously. Sanger sequencing suggested that the c. 853del (p.Ala285Leufs*24) variant has derived from his deceased mother, whilst the c. 1276A>T (p.Lys426X) variant has derived from his father. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), both variants were classified as pathogenic (PVS1+ PM2_Supporting+ PP4). Conclusion:The compound heterozygous variants of the PMFBP1 gene probably underlay the Acephalic spermatozoa syndrome in this patient. The discovery of the novel variants has also enriched the mutational spectrum of Acephalic spermatozoa syndrome.

OBJECTIVE: To explore the clinical feature and gene variant for two cases of primary male infertility caused by severe asthenospermia and to analyze the etiology of the disease. METHODS: Genomic DNA of peripheral blood samples of patients and their parents was extracted and gene variant analysis of the patients was conducted by using whole exome sequencing. Suspected pathogenic variant was verified by Sanger sequencing and pathogenic analysis. RESULTS: Whole exome sequencing showed that the DNAH1 gene of patient 1 had two heterozygous variants of c.2016T>G(p.Y672X) and c.6017T>G (p.V2006G). The DNAH1 gene of patient 2 had a homozygous variant of c.2610G>A(p.W870X), which were inherited from his father and mother, respectively. According to American College of Medical Genetics and Genomics standards and guidelines, the c.2016T>G (p.Y672X) and c.2610G>A (p.W870X) varaints of DNAH1 gene were predicted to be pathogenic (PVS1+PM2+PM3+PP3). CONCLUSION The two patients of multiple morphological abnormalities of the sperm flagella may be caused by DNAH1 gene variant, which has resulted in primary male infertility.
Dyneins/genetics* ; Genomics ; Humans ; Infertility, Male/genetics* ; Male ; Mutation ; Sperm Tail/pathology* ; Whole Exome Sequencing

To unravel the genetic mechanisms of disease and physiological traits, it requires comprehensive sequencing analysis of large sample size in Chinese populations. Here, we report the primary results of the Chinese Academy of Sciences Precision Medicine Initiative (CASPMI) project launched by the Chinese Academy of Sciences, including the de novo assembly of a northern Han reference genome (NH1.0) and whole genome analyses of 597 healthy people coming from most areas in China. Given the two existing reference genomes for Han Chinese (YH and HX1) were both from the south, we constructed NH1.0, a new reference genome from a northern individual, by combining the sequencing strategies of PacBio, 10× Genomics, and Bionano mapping. Using this integrated approach, we obtained an N50 scaffold size of 46.63 Mb for the NH1.0 genome and performed a comparative genome analysis of NH1.0 with YH and HX1. In order to generate a genomic variation map of Chinese populations, we performed the whole-genome sequencing of 597 participants and identified 24.85 million (M) single nucleotide variants (SNVs), 3.85 M small indels, and 106,382 structural variations. In the association analysis with collected phenotypes, we found that the T allele of rs1549293 in KAT8 significantly correlated with the waist circumference in northern Han males. Moreover, significant genetic diversity in MTHFR, TCN2, FADS1, and FADS2, which associate with circulating folate, vitamin B12, or lipid metabolism, was observed between northerners and southerners. Especially, for the homocysteine-increasing allele of rs1801133 (MTHFR 677T), we hypothesize that there exists a "comfort" zone for a high frequency of 677T between latitudes of 35-45 degree North. Taken together, our results provide a high-quality northern Han reference genome and novel population-specific data sets of genetic variants for use in the personalized and precision medicine.

Objective To explore the role of D-dimer (D-D) level in patients with acute exacerbation of chronic obstructive disease (AECOPD) in predicting the prognosis of patients.Methods 99 patients with AECOPD in our hospital from July 2008 to December 2010 were selected as the subjects,and were divided into observation group (n=50) and control group (n=49) according to the expression of D-D in patients with positive (D-D level cutoff value of 500 g/L).The basic information and survival of the two groups of patients were investigated,and the level of serum D-D was detected by scatter turbidimetry.The receiver operating characteristic curve (ROC) was used to evaluate the predictive value of D-D on the survival of the patients.The survival time of the two groups was compared by Kaplan-Meier.Results There was no statistically significant difference in gender,age and GOLD score between the two groups (P＞0.05).The level of D-D,the time of first hospitalization,the number of re hospitalization,the first hospital mortality and re hospitalization mortality in the observation group were significantly higher than those in the control group (P＜0.05).The ROC curve showed that the truncated value of D-D was 1 171.34 g/L and 893 g/L,respectively.The average survival time of patients in the observation group was 30.37 months and the control group was 46.37 months,respectively;Kaplan-Meier study showed that the survival time of patients in the observation group was significantly lower than that in the control group (x2 =6.371,P＜0.05).Conclusion The level of D-D in patients with AECOPD is of great value in predicting the prognosis of patients,which can be used as an independent indicator of disease progression.

Objective:To investigate the effect of continuous blood purification (CBP) on inflammatory factors and immune function in patients with sepsis,in order to provide reference for clinical treatment of sepsis.Methods:A total of 76 patients with severe sepsis,who were treated in ICU of Daqing Oilfield General Hospital from June 2014 to December 2016,were selected.The patients were divided into conventional group (n=36) and CBP group (n=40) according to the received treatment method.The patients in the conventional group were treated with targeted therapy and life support,while the patients in the CBP group were treated with CBP on the basis of targeted therapy.The levels of interleukin-6 (IL-6),interleukin-10 (IL-10),tumor necrosis factor-α (TNF-α) and C reactive protein (CRP) of the patients in the two groups were detected and compared before treatment and after 3d of treatment.The CD3+,CD4+,CD8+T cell and the CD4+/CD8+ ratio before treatment and after 3 d of treatment were calculated and compared between the two groups.Results:There was no significant difference in the level of serum inflammatory factors between the two groups before treatment (P＞0.05).The levels of serum inflammatory factors in the two groups were significantly decreased after 3 d of treatment,and the levels of IL-6,TNF-α and CRP in the CBP group were significantly lower than those in the conventional group (P＜0.05).There was no significant difference in immune function indexes between the conventional group and the CBP group before treatment (P＞0.05).After 3d of treatment,CD3+,CD4+ and CD4+/CD8+ ratio were significantly increased in the two groups,and the CD4+ and CD4+/CD8+ ratio of the CBP group were sig-nificantly higher than those of the conventional group (P＜0.05).Conclusion:The application of CBP in the treatment of patients with sep-sis can obviously improve the expression of inflammatory factors of patients,and can improve the immune function and promote the re-covery of immune function.

Objective To prepare butenafine hydrochloride plastics,investigate the prescription composition and make a quality control standard for the preparation.Methods Film-forming time and appearance quality as the evaluation index,the quality control standard of butenafine hydrochloride according to the Chinese pharmacopoeia two section ( 2010 edition ) was made.ResuIts The prescription of butenafine hydrochloride plastics was identified as:1%butenafine hydrochloride(w/w),10%glycerol(w/w),3%carbomer 971PNF(w/w),0.1% ethyl p-hydroxybenzoate(w/w),moderate anhydrous sodium sulfite(pH adjusting agent) and 95% ethanol (solution).The preparation was colorless,transparent and viscous semi-solid with pH4.5.A content determination method of butenafine hydrochloride with HPLC was established and the result was stable and reliable .ConcIusion The butenafine hydrochloride has several advantages such as preparation simply , stable property,application convenience and quality control.It is a potential preparation to develop.

Objective To investigate effect of high glucose on the function of endothelial and the underlying mechanisms in human umbilical vascular endothelial cells (HUVECs).Methods The experiment was divided into 4 groups: normal group (NG), low dose group (LG), middle dose group (MG) and high dose group ( HG) .The concentration of glucose in the culture medium was 5.5, 10, 20, 30 mmol/L in the 4 groups, respectively.The HUVECs was cultured for 0, 24, 48 h.At different time point, the cell viability were measured by MTT.The secretary content of nitric oxide (NO) in the supernatant were detected using test kit.The extraction of protein were extracted for Western blot analysis to detect the expression of endothelial nitric oxide synthase (eNOS).ResuIts Compared with normal group at same time point (cultured for 24 h), the cell viability and the content of NO were significantly decreased in LG, MG(P<0.05).The expression of eNOS in HG were markedly reduced (P<0.01).Compared with normal group at same time point (cultured for 48 h), the cell viability decreased significantly in HG (P <0.05).The expression of eNOS were markedly decreased 11.91, 25.72 and 34.50% in LG, MG and HG, respectively.A rising trend of cell viability were found in NG, LG and MG, but a descending trend were found in HG within 48 h.ConcIusion The cell viability were significantly affected by high glucose.The endothelial dysfunction induced by high glucose may be associated with the reduction of eNOS and NO production.

Objective To determine serum concentration of methotrexate in children with acute lymphoblastic leukemia by HPLC and explore the application practice.Methods The separation was performed in a Wondasil C18-WR column (4.6 mm ×150 mm, 5 μm) with a mobile phase of methanol-0.15%phosphoric acid solution(21:79 ) and determined at 306 nm.The sample of serum was centrifuged after protein precipitation with perchloric acid.Detected results for children to establish monitoring files.ResuIts The linearity was well at 0.02 ~50 μg/mL of methotrexate ( r =0.9990,n=6).The average recovery was 96.50%, RSD of within day and between days were less than 15%.Established monitoring files for six children and after 3 courses of treatment, the blood concentration of methotrexate in 4 children were small and they were in a safe range , which could reduce the frequency of monitoring.ConcIusion This method is simple, sensitive and accurate for methotrexate detection in serum.The children with stable concentration and within safety range, can reduce monitioring frequency.It could bring the children less pain and relieve the burden of family.

Objective To prepare pemirolast potassium nasal spray which could be used in treatment of allergic rhinitis and make a quality control standard for the preparation.Methods The preparation materials of nasal spray was selected which could meet the physical and chemical properties of pemirolast potassium as well as the characteristics of intranasal administration.The quality control standards for pemirolast potassium nasal spray was made according to the nasal spray quality standards for the formulation pH, property, loading capacity, total bottle spray times, puff volume, sedimentation volume ratio and detection of content of “Chinese Pharmacopoeia” two section (2010 edition).Results The formulation prescription of pemirolast potassium nasal spray was:0.1% potassium pemirolast, 0.8% CMC-Na, 0.01% benzalkonium bromide, glacial acetic acid ( pH adjusting agent) and the solution was 5% mannitol.The formulation was a white suspension, pH=7.The mean value of installed capacity was (25.12 ±0.16) mL and installed capacity of each bottle was above 23.75 mL(95%); total spray times of each bottle was above 160 sprays and spray volume was above 0.140 0 g/press; sedimentation volume ratio was 1, the above testing programmes of quality standard met the requirements and were controlled.The standard curve of pemirolast potassium reference substance was Y=81085X+15264(r=0.999 8), linearity range was 0.5~25.0μg/mL.The average recovery rate of pemirolast potassium sample was 99.39%, and precision met the requirements.The pemirolast potassium content of test sample was 99.2% of labeled amount.Conclusion The formulation prescription of pemirolast potassium nasal spray is suitable and the preparation is possessed several advantages such as stability, well-distributed spray, application convenience and quality controlled.Therefore, nasal administrated formulation of pemirolast potassium is potentially in further.

Purpose To investigate the clinical and pathological features of inflammatory fibroid polyps ( IFPs) and to discuss its diag-nosis, differential diagnosis and treatment. Methods Clinicopathological data of 14 cases of IFPs were collected and analyzed retro-spectively. Results Most of the 14 cases were found to have submucosal polyps of the digestive tract protruding growing into the cavi-ty. Perivascular onion skinning which formed by spindle fibrocytes around vascular proliferation and eosinophilic inflammatory infiltrates were present as the characteristic feature of lesions. ABC results demonstrated that the lesions showed fibroblastic, myofibroblastic and dendritic cell signs. Conclusions IFPs are rare, benign tumors that can arise throughout the digestive tract, with unique histological changes in pathology. Surgical excision or endoscopic removal is the main treatment.