Abstract: Objective　To detect and analyze the antiserum of Yersinia pestis phage in Marmota himalayana blood from the natural plague foci of Qinghai-Tibet Plateau by micro-bolus technique, to provide a theoretical basis for interaction between phages and mammalian immunology, phage therapy and interaction between bacteriophage and ecology in future. Methods　Using diagnostic Yersinia pestis phage and 3 wild plague phages from Qinghai-Tibet Plateau Natural Plague Foci as antigens, 847 serums of Marmota Himalayana blood, from Tongde, Guinan, Gonghe, Xinghai, Tianjun foci counties in Qinghai Plateau, were collected from July to September in 2020, 2021 and determined on antiserum of Yersinia pestis phage by microplate method and double agar plate method. Results　The neutralization reaction experiment lasted for 24 hours between 4 phage and 847 serums by microplate method independently. These mixtures were tested by double agar plate method. All results were negative on antiserum of Yersinia pestis bacteriophage. Conclusions　The positive antiserum of Yersinia pestis phage in Marmota himalayana were not found the natural plague foci of Qinghai-Tibet Plateau, which agreed with plague epidemiology in 5 foci counties in Qinghai plateau from 2020-2021, that was a characteristic of the resting period. In other words, it was in the absence of plague pathogen. It also showed indirectly that the absence or weak presence of Yersinia pestis bacteriophage in the plague foci. It showed a lower frequency on host animals coming into contact with phages naturally. The antiserum of Yersinia pestis phage may be related to the form of plague infection and the intensity of the disease.

Objective To investigate the clinical, cerebrospinal fluid and imaging characteristics of 5 cases of diffuse meningeal melanomatosis.Methods The clinical manifestation, features of cerebrospinal fluid and image of 5 patients with meningeal melanomatosis diagnosed by biopsy or autopsy were retrospectively summarized.Results Clinical manifestations of these 5 cases included intracranial hyperpressure, meningeal irritation sign, intracranial nerves impairment, root pain of spinal nerve.In all of these 5 cases, retina hyperpigmentation above left discus opticus was found by funduseope in one case, and congenital melanocytic nevi were found in 4 patients, in which 2 cases were giant congenital melanocytic nevi.Increased lumbar puncture cerebrospinal fluid(CSF)pressure occurred in all cases.Subarachnoid hemorrhage was found in 3 cases.Analysis of CSF revealed increased protein in 4 cases and decreased glucose in 3 cases.Cranial MRI obtained after the intravenous administration of Gd-DTPA showed leptomeningeal enhancement.Malignant melanoma cells were found in CSF of 3 cases.Metastatic malignant melanoma cells were found by biopsy of axillary fossa lymph node in one case.Autopsy of one case revealed diffuse black pigmentation of the leptomeninges, especially in base of skull.Two cases were diagnosed as metastatic meningeal melanomatosis and 3 cases were possible primary meningeal melanomatosis. Conclusions Menings, root of cranial nerve and spinal nerve are impaired in meningeal melanomatosis, which is usually accompanied by congenital melanocytic nevi.Subarachnoid hemorrhage implies meningeal melanomatosis.Diagnosis can be identified when malignant melanoma cells are found in CSF.

OBJECTIVETo appraise the effect of single- and two-layer Percoll density gradient centrifugation in sperm separation.

METHODSTwenty semen specimens underwent single-(50%) and two-layer (90% and 45%) density gradient centrifugation, respectively. The sperm class analyzer (SCA) was used to analyze sperm density, motility and dynamic parameters and round cell density before and after the treatment.

RESULTSAfter separation, the sperm recovery rate of the single-layer method was (65.5 +/- 12.8)%, significantly higher than that of the two-layer method (P < 0.01). The percentages of grade a sperm of the single- and two-layer method were significantly higher than pre-treatment (P < 0.05, P < 0.01), that of the single-layer was significantly lower than that of the two-layer method (P < 0.05), but the percentage of grade c sperm of the former was significantly higher than that of the latter (P < 0.05). Compared with pre-treatment, the percentage of grade a + b sperm of the two-layer method was significantly higher (P < 0.05), while that of the single-layer method showed no significant difference (P > 0.05), and the round cell density of both the methods was significantly lower (P < 0.05, P < 0.01), with no significant differences between the two methods (P > 0.05).

CONCLUSIONThe single-layer method yields a higher rate of sperm recovery and causes little change in the sperm motility, while the two-layer method effects a lower rate and significantly improves sperm motility. Both the methods can efficiently separate sperm from round cells, and each has its own advantages and its application value in in vitro treatment of sperm.

Cell Separation ; methods ; Centrifugation, Density Gradient ; methods ; Humans ; Male ; Povidone ; Silicon Dioxide ; Sperm Count ; methods ; Spermatozoa ; cytology

Objective :To study correlation between treadmill exercise test assessing myocardial ischemia and coronary stenosis .Methods :A total of 150 patients with stable angina pectoris in our hospital ,according to vascular stenosis by coronary CT were divided into group A (stenosis＜ 50%,n＝ 50) ,group B (stenosis 50% ～75%,n＝ 50) and group C (stenosis ＞75%,n＝50).Coronary stenotic rate and ST depression during exercise test were compared a-mong three groups.Spearman method was used to analyze correlation between treadmill exercise positive rate and coronary stenosis .Results :Compared with group B ,there were significant rise in coronary diameter stenotic rate [(63.64 ± 4.21)% vs.(66.71 ± 4.46)%] and vascular diameter of reference segment [(2.92 ± 0.23) mm vs.(3.03 ± 0.21) mm] in group C ,P＜0.05 or ＜0.01. Along with coronary stenosis aggravated ,there was significant re-duction in onset time of ST depression [(712.3 ± 202.7) s vs.(602.3 ± 210.4) s vs.(501.2 ± 236.1) s] ,and signif-icant rise in duration [ (425.4 ± 200.5) s vs.(520.8 ± 205.8) s vs.(603.4 ± 198.4) s] and treadmill exercise posi-tive rate (64.00% vs.82.00% vs .92.00%) ,in group A ,B ,C respectively ;and ST depression extent of group C was significantly higher than that of group A [(2.4 ± 1.1) mV vs.(1.9 ± 0.8) mV] ,P＜0.05 or ＜0.01. Spearman correlation analysis indicated that treadmill exercise positive rate (myocardial ischemia rate ) was not significantly correlated with coronary stenotic degree (r＝3.425 , P＝0.126).Conclusion :Although treadmill exercise test pos-sesses important significance in diagnosis of myocardial ischemia ,but coronary stenotic degree is not correlated with myocardial ischemic degree .which should raise clinical attention

Objective: To evaluate the feasibility, safety, and effectiveness of CT-guided microcoil localization of solitary pulmonary nodules (SPNs) for guiding video-assisted thoracoscopic surgery (VATS). Materials and Methods: Between June 2016 and October 2019, 454 consecutive patients with 501 SPNs who received CTguided microcoil localization before VATS in our institution were enrolled. The diameter of the nodules was 0.93 ± 0.49 cm, and the shortest distance from the nodules to the pleura was 1.41 ± 0.95 cm. The distal end of the microcoil was placed less than 1 cm away from the nodule, and the proximal end was placed outside the visceral pleura. VATS was performed under the guidance of implanted microcoils without the aid of intraoperative fluoroscopy. Results: All 501 nodules were marked with microcoils. The time required for microcoil localization was 12.8 ± 5.2 minutes. Microcoil localization-related complications occurred in 179 cases (39.4%). None of the complications required treatment. A total of 463 nodules were successfully resected under the guidance of implanted microcoils. VATS revealed 38 patients with dislocated microcoils, of which 28 underwent wedge resection (21 cases under the guidance of the bleeding points of pleural puncture, 7 cases through palpation), 5 underwent direct lobectomy, and the remaining 5 underwent a conversion to thoracotomy. In 4 cases, a portion of the microcoil remained in the lung parenchyma. Conclusion CT-guided microcoil localization of SPNs is safe and reliable. Marking the nodule and pleura simultaneously with microcoils can effectively guide the resection of SPNs using VATS without the aid of intraoperative fluoroscopy.

Objective: To evaluate the feasibility, safety, and effectiveness of CT-guided microcoil localization of solitary pulmonary nodules (SPNs) for guiding video-assisted thoracoscopic surgery (VATS). Materials and Methods: Between June 2016 and October 2019, 454 consecutive patients with 501 SPNs who received CTguided microcoil localization before VATS in our institution were enrolled. The diameter of the nodules was 0.93 ± 0.49 cm, and the shortest distance from the nodules to the pleura was 1.41 ± 0.95 cm. The distal end of the microcoil was placed less than 1 cm away from the nodule, and the proximal end was placed outside the visceral pleura. VATS was performed under the guidance of implanted microcoils without the aid of intraoperative fluoroscopy. Results: All 501 nodules were marked with microcoils. The time required for microcoil localization was 12.8 ± 5.2 minutes. Microcoil localization-related complications occurred in 179 cases (39.4%). None of the complications required treatment. A total of 463 nodules were successfully resected under the guidance of implanted microcoils. VATS revealed 38 patients with dislocated microcoils, of which 28 underwent wedge resection (21 cases under the guidance of the bleeding points of pleural puncture, 7 cases through palpation), 5 underwent direct lobectomy, and the remaining 5 underwent a conversion to thoracotomy. In 4 cases, a portion of the microcoil remained in the lung parenchyma. Conclusion CT-guided microcoil localization of SPNs is safe and reliable. Marking the nodule and pleura simultaneously with microcoils can effectively guide the resection of SPNs using VATS without the aid of intraoperative fluoroscopy.

AIMTo explore the possible effect of the deleted in azoospermia (DAZ) copy cluster deletion on spermatogenesis in the Chinese population, the deletion of the azoospermia factor c (AZFc) region was analyzed in 346 normozoospermic men.

METHODSThree DAZ single nucleotide variant loci and seven AZFc-specific sequence-tagged sites were examined with polymerase chain reaction (PCR)-restriction fragment length polymorphism and routine PCR.

RESULTSFive (1.4%) of the normozoospermic men were found to have deletion of gr/gr-DAZ1/DAZ2. None of the men were found to have b2/b4-entire DAZ deletion.

CONCLUSIONThe presence of gr/gr-DAZ1/DAZ2 deletion in five men with normozoospermia suggests that this deletion per se may not be sufficient for spermatogenic impairment in Chinese men.

Adult ; Asian Continental Ancestry Group ; genetics ; China ; Chromosomes, Human, Y ; genetics ; Gene Deletion ; Humans ; Male ; Oligospermia ; genetics ; Polymorphism, Restriction Fragment Length ; RNA-Binding Proteins ; genetics ; Recombination, Genetic ; genetics ; Sequence Tagged Sites ; Spermatogenesis ; genetics

OBJECTIVETo express the cloned gene glycoprotein I (gpI) of varicella-zoster virus (VZV), Beijing VZV 84-7 strain in insect cells and to purify its expression product.

METHODSThe gene coding for gpI of VZV was amplified from viral DNA by PCR and cloned into baculovirus transfer vector (pBacPAK9), and recombinant transfer vector plasmid pBacVZVgpI was obtained. The inserted gpI gene in the pBacVZVgpI was sequenced. Insect cells Sf 9 were co-transfected with the recombinant transfer vector plasmid pBacVZVgpI and wild type linear baculovirus BacPAK6 (digested with Bsu36I) DNA. The recombinant baculoviruses containing the VZV 84-7 gpI gene was isolated through several rounds of limited dilution. Recombinant protein gpI was expressed in insect cells Sf 9, postinfected with recombinant baculoviruses. The expressed recombinant gpI was purified by lectin affinity chromatography and its antigenicity and immunogenicity were investigated.

RESULTSThe gene coding for gpI of VZV was obtained by PCR and the gpI gene of pBacPAK9 was confirmed by DNA sequencing. The recombinant gpI was expressed in insect cells Sf 9, post-infected with recombinant baculovirus and identified by SDS-PAGE and western blotting, with its product in cell culture reaching the peak in 72 hours and with a molecular mass of 58 kd and 70 kd, the same as theoretical values. Results of immunoassay with cell lysates infected by recombinant baculoviruses indicated that recombinant protein expressed in insect cells had ability of eliciting specific antibodies against native VZV in mice and complement-dependent neutralizing antibodies. The purified recombinant gpI gave a product with a purity of more than 80%. ELISA and Western-blot analysis demonstrated that purified protein had specific VZV antibody-binding activity. This suggested that the recombinant gpI expressed in insect cells had the same biological characteristics as its native counterpart.

CONCLUSIONBaculovirus-insect cells could be used to express the gene of VZV gpI, which could provide a basis for quantitative analysis of VZV antigen, and preparation of its subunit vaccine.

Animals ; Cell Line ; DNA, Viral ; genetics ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; genetics ; Genetic Vectors ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; isolation & purification ; Spodoptera ; cytology ; genetics ; Viral Envelope Proteins ; genetics ; isolation & purification

Objective To evaluate the protective effect of Yersinia pestis capsular antigen F1 and recombinant rV270 on mice after immunization with them.Methods According to body weight,40 female Balb/c mice aged 6 to 8 weeks were randomly divided into four experimental groups(Fl-10 μg + aluminum adjuvant,F1-20 μg + aluminum adjuvant,rV-10 μg + aluminum adjuvant,and rV-20 μg + aluminum adjuvant) and a control group,8 in each group.Mice in experimental groups were immunized with the natural antigen F1 and recombinant antigen rV270 adsorbed to 25％ aluminum adjuvant and the control group was immunized with the same amount of aluminum adjuvant.Each mouse was immunized at the hind leg muscle with 100 ml immunizing agent,then a booster immunization was done once on the 21st day after the first immunization.The blood of all mice was collected on the 8th week after the first immunization,serum antibody titers were detected by ELISA and the data of antibody titers were analyzed by t test for comparison between groups.At the same time the mice were injected subcutaneously with 2000-fold LD50 of Yersinia pestis virulent strain 141,after 14 days,the protective effect of immunization was analyzed.Results The control group did not produce antibody.Antibody geometric mean titers (GMT) of the F1-10 mg + aluminum adjuvant and F1-20 mg + aluminum adjuvant groups were 1 ∶ 30443.9,and 1 ∶21527.8,respectively,and compared between the two groups,the difference was not statistically significant (t =1.1282,P ＞ 0.05).The GMTs of the rV-10 μg + aluminum adjuvant and rV-20 μg + aluminum adjuvant groups were 1 ∶ 13957.3 and 1 ∶18100.9,respectively,and compared between the two groups,the difference was not statistically significant(t =0.9408,P ＞ 0.05 ).After subcutaneous injection with Yersinia pestis virulent strain 141,all mice died in the control group but all survived in the experimental group.Conclusion The immune activity of natural antigen F1 and recombinant rV270 is high,which can be used as the main component of subunit vaccine in the plague subunit vaccine study.

In the research community, resistance to apoptosis is often considered a hallmark of cancer. However, pathologists who diagnose cancer via microscope often see the opposite. Indeed, increased apoptosis and mitosis are usually observed simultaneously in cancerous lesions. Studies have shown that increased apoptosis is associated with cancer aggressiveness and poor clinical outcome. Furthermore, overexpression of Bcl-2, an antiapoptotic protein, is linked with better survival of cancer patients. Conversely, Bax, CD95, Caspase-3, and other apoptosis-inducing proteins have been found to promote carcinogenesis. This notion of the role of apoptosis in cancer is not new; cancer cells were found to be short-lived 88 years ago. Given these observations, resistance to apoptosis should not be considered a hallmark of cancer.
Animals ; Apoptosis ; physiology ; Biomarkers, Tumor ; metabolism ; Carcinogenesis ; metabolism ; Caspase 3 ; metabolism ; Humans ; Lymphoma, B-Cell ; metabolism ; pathology ; Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Treatment Outcome ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism