Objective:To explore the genetic basis of a myopathic patient with pathological characteristics including tubular aggregates and vacuoles.Methods:Next generation sequencing was carried out for the patient, and candidate variant was verified by Sanger sequencing.Results:Genetic testing revealed that the patient has harbored a heterozygous c. 730G>C (p.D244H) variant of Calsequestrin 1 ( CASQ1) gene. The same variant was not found in his unaffected parents. Based on guidelines from the American College of Medical Genetics and Genomics, the variant was rated as pathogenic (PS1+ PM2+ PP3). Conclusion:The novel c. 730G>C (p.D244H) variant of the CASQ1 gene probably underlay the myopathy in this patient. Above finding has enriched the mutational spectrum of the CASQ1 gene.

italic>O-methyltransferases (OMTs) are one of the key tailoring enzymes in the biosynthesis of many natural products, O-methylation can not only reduce the reactivity of natural products, but also alter their solubility, stability and biological activities. Based on the transcriptome data of Ardisia japonica, a full-length cDNA sequence of candidate OMT (termed as AjOMT1) was cloned by reverse transcription-polymerase chain reaction (RT-PCR) and expressed in Escherichia coli (E. coli) for the first time. In vitro enzyme catalytic activity assay showed that the recombinant AjOMT1 could effectively catalyze quercetin to form O-methylated products. Most importantly, AjOMT1 showed unprecedented substrate promiscuity towards structurally various compounds including flavonoids, stilbenes, coumarins, alkaloids and phenylpropanoids, especially preferring to the compounds with adjacent phenolic hydroxyl groups, and showed regio-selectivity. These results suggested that AjOMT1 could be used as the tool enzyme to conduct O-methylation of different types of compounds to produce diverse active methylated products, and provide a new method for drug discovery, even universal part for synthetic biology of natural products.

Objective:To summarize the clinical manifestations, electrophysiological, muscle magnetic resonance imaging (MRI), pathological, and genetic characteristics of 8 patients with Emery-Dreifuss muscular dystrophy (EDMD) to improve the recognition and diagnosis of EDMD.Methods:Eight patients with EDMD confirmed by gene analysis admitted to Hebei Medical University Third Hospital from 2011 to 2022 were enrolled. The detailed clinical symptoms, neurophysiological examination, electrophysiological changes (electromyography and electrocardiography), skeletal muscle MRI characters, skeletal muscle pathological features and gene mutations were analyzed retrospectively.Results:The age of onset ranged from 2.0 to 6.0 (3.6±1.2) years. All patients had insidious onset and progressive development. Muscle weakness was the first symptom for 7 cases that manifested as difficulty in squatting and walking up stairs. Later, spinal ankylosis and joint contracture occurred. One patient had scoliosis as the first symptoms. Abnormal electrocardiogram was found in 4 cases. The electromyography of all patients showed myogenic damage. Muscle biopsy demonstrated dystrophic features in 1 patient, and other myopathic features, including a variation in muscle fiber size, a marked increase in internal nuclei, and, smaller diameter of typeⅠfibers. Next-generation sequencing result showed that 6/8 cases carried 4 LMNA heterozygous mutations (c.1583C>G, c.1357C>T, c.148C>T, c.1336A>G); 1/8 case carried EMD hemizygous mutation (c.501C>G); 1/8 carried SYNE1 heterozygous mutation (c.4364G>A). Conclusions:EDMD has highly clinical and genetical heterogeneity. The onset age is usually in childhood. The first symptom is characterized by weakness of lower limbs and abnormal walking posture. Electromyography shows myogenic lesion. Skeletal muscle MRI shows selective fat infiltrations. Muscle biopsy pathology lacks characteristic pathological findings. It is difficult to make diagnosis and differential diagnosis by clinical manifestations and auxiliary examination in the early stage of the disease. The second generation sequencing technology can improve the early diagnosis rate of EDMD.

Objective: To investigate the distribution of HIV-1 genetic subtypes and pretreatment drug resistance (PDR) among men who have sex with men (MSM) from 19 cities of 6 provinces in China. Methods: From April to November 2019, 574 plasma samples of ART-naive HIV-1 infected MSM were collected from 19 cities in Hebei, Shandong, Jiangsu, Zhejiang, Fujian, and Guangdong provinces, total ribonucleic acid (RNA) was extracted and amplified the HIV-1 pol gene region by nested polymerase chain reaction (PCR) after reverse transcription. Then sequences were used to construct a phylogenetic tree to determine genetic subtypes and submitted to the Stanford drug resistance database for drug resistance analysis. Results: A total of 479 samples were successfully amplified by PCR. The HIV-1 genetic subtypes included CRF01_AE, CRF07_BC, B, CRF55_01B, CRF59_01B, CRF65_cpx, CRF103_01B, CRF67_01B, CRF68_01B and unrecognized subtype, which accounted for 43.4%, 36.3%, 6.3%, 5.9%, 0.8%, 0.8%, 0.4%, 0.4%, 0.2% and 5.5%, respectively. The distribution of genetic subtypes among provinces is statistically different (χ²=44.141, P<0.001). The overall PDR rate was 4.6% (22/479), the drug resistance rate of non-nucleoside reverse transcriptase inhibitors, nucleoside reverse transcriptase inhibitors, and protease inhibitors were 3.5% (17/479), 0.8% (4/479) and 0.2% (1/479), respectively. The PDR rate of recent infections was significantly higher than that of long-term infections (χ²=4.634, P=0.031). Conclusions: The HIV-1 genetic subtypes among MSM infected with HIV-1 from 19 cities of 6 provinces in China are diverse, and the distribution of subtypes is different among provinces. The overall PDR rate is low, while the PDR rate of recent infections was significantly higher than that of long-term infections, suggesting the surveillance of PDR in recent infections should be strengthened.
China/epidemiology* ; Cities ; Drug Resistance ; Drug Resistance, Viral/genetics* ; Female ; Genotype ; HIV Infections/epidemiology* ; HIV Seropositivity/drug therapy* ; HIV-1/genetics* ; Homosexuality, Male ; Humans ; Male ; Phylogeny ; Reverse Transcriptase Inhibitors/therapeutic use* ; Sexual and Gender Minorities

Applying Chinese medicine (CM) is an important strategy for malignant tumor treatment in China. One of the significant characteristics of CM is to treat diseases based on syndrome differentiation. For Western medicine, it is of important clinical significance to formulate guidelines for the diagnosis and treatment of cancer patients based on the characteristics of disease differentiation. In Chinese clinical practice, the combination of disease differentiation and syndrome differentiation is an important feature for cancer treatment in the past. Currently, molecular profiling and genomic analysis-based precision medicine optimizes the anticancer drug design and holds the greatest success in treating cancer patients. Therefore, we want to know which populations of cancer patients can benefit more from CM treatment if the theory of precision medicine is applied to CM clinical practice. So, we developed a novel diagnostic and therapeutic strategy "disease-syndrome differentiation-genomic profiling-prescriptions" for cancer patients by CM syndrome differentiation and precision medicine. As a result, this strategy has greatly enhanced the anti-tumor efficacy of CM and improved clinical outcomes for cancer patients with some gene mutations. Our idea will hopefully establish a novel approach for the inheritance and innovation of CM.
Antineoplastic Agents ; Drugs, Chinese Herbal/therapeutic use* ; Humans ; Medicine, Chinese Traditional ; Neoplasms/therapy* ; Precision Medicine ; Syndrome

Objective:To study the clinical characteristics and etiological distribution characteristics of plastic bronchitis in children, analyze its early warning indicators, and evaluate the clinical diagnosis and treatment effect of flexible bronchoscopy.Methods:The clinical data of 232 children with severe pneumonia admitted to Guiyang Maternal and Child Health Hospital from January 2019 to February 2021 were retrospectively analyzed.The children were divided into the plastic bronchitis group and non-plastic bronchitis group according to bronchoscopic results.The gender, age, clinical manifestations, auxiliary examinations, imaging features, bronchoscopy findings and treatment of the children were collected, compared and analyzed, comparison between two groups by t test and χ2 test. Results:A total of 232 children were included in this study, including 98 cases in the plastic bronchitis group and 134 cases in the non-plastic bronchitis group.The main symptoms of both groups were fever, cough and shortness of breath.The age of onset in the plastic bronchitis group was (54.640±37.085) months, and the age of onset in the non-plastic bronchitis group was (14.870±19.813) months.The difference in the age of onset between the two groups was statistically significant ( t=9.656, P<0.001). The average hospitalization days of the plastic and non-plastic bronchitis groups were (16.133±6.227) d and (12.690±4.287) d, respectively.Significant difference was found in the average hospitalization days between the two groups ( t=4.721, P<0.001). The average fever days of the plastic bronchitis group were (10.090±3.473) d, and the average fever days of the non-plastic bronchitis group were (6.030±4.850) d. There was significant difference in the average fever days between the two groups ( t=5.654, P<0.001). The age of onset, hospitalization days, and fever days of the plastic bronchitis group were larger than those of the non-plastic bronchitis group (all P<0.001). The physical examination suggested that 40% (39/98) of patients in the plastic bronchitis group had reduced the breath sounds, and this percentage was significantly higher than that in the non-plastic bronchitis group[6%(8/134)]. The plastic bronchitis group had lower partial pressure of blood oxygen (PO 2) and oxygen saturation (SO 2) levels than the non-plastic bronchitis group (all P<0.01). The plastic bronchitis group had a higher percentage of neutrophils (N), C-reactive protein (CRP) level, procalcitonin (PCT) level, lactate dehydrogenase (LDH) level and D-dimer level than the non-plastic bronchitis group (all P<0.01). According to the imaging results, in the plastic bronchitis group, lung consolidation was found in 72 cases (73%, 72/98), atelectasis in 32 cases (33%, 32/98), and pleural effusion in 33 cases (34%, 33/98). In the non-plastic bronchitis group, 65%(87/134) cases had lung consolidation, 5%(7/134) cases had atelectasis, 3.7% (5/134) cases had pleural effusion.The first pathogen detected in 46.9% of the patients in the plastic bronchitis group was Mycoplasma pneumoniae (MP), and the percentage was significantly higher that in the non-plastic bronchitis group (11.1%). Flexible bronchoscopy was performed on both groups at their admission.The plastic bronchitis group received the flexible bronchoscopy check for (2.960±1.157) times on average, and the non-plastic bronchitis group was tested for (1.140±0.371) times on average.Of 98 children in the plastic bronchitis group, 95 cases were improved and discharged, 2 cases were transferred, and 1 case died.All 134 children in the non-plastic bronchitis group were improved and discharged. Conclusions:Preschool and school-age children, fever ≥10 d, PCT, CRP, LDH, D-dimer levels are early warning signs of plastic bronchitis clinically.MP is still the primary pathogen causing plastic bronchitis.Flexible bronchoscopy technique is a key measure for timely diagnosis and effective treatment of plastic bronchitis.

Objective:To summarize the infective characteristics of Fournier's gangrene (FG) and evaluate the effect of negative pressure wound therapy (NPWT).Methods:A retrospective case control study was conducted to analyze the clinical data of 31 patients with FG admitted to Peking University First Hospital from May 2010 to September 2020, including 29 males and 2 females, aged 21-78 years [(55.2±2.0)years]. A total of 29 patients were caused by infectious diseases of the perianal and urinary system, and the rest two patients were caused by vulvar infection and retroperitoneal abscess. A total of 23 patients were treated with NPWT (Group A) and 8 patients were treated with conventional dressing (Group B). Characteristics of pathogen, drug-resistance rate, medical treatment and prognosis for all patients were summarized. The hospitalization duration, numbers of operation and wound healing time were compared between two groups.Results:Monomicrobial infection was identified in 14 patients, while polymicrobial infection in 15 patients, fungal infection in 1 and culture-negative in 1. Escherichia coli, Enterococcus faecalis, Enterococcus faecium, Klebsiella pneumoniae and Staphylococcus haemolyticus were the most common pathogenic bacteria. The resistance rate of gram-negative bacilli to third-generation cephalosporins was 37%. Staphylococcus haemolyticus were methicillin-resistant Staphylococcus. The carbapenem antibiotics combined with vancomycin antibiotics were used for all patients as the empirical anti-infection treatment. Three patients died, and the rest 28 patients were followed up for 3 to 12 months [(10.8±2.6)months] after discharge. All the wounds were healed well without recurrence. In Group A and Group B, the hospitalization duration was (37.4±15.0)days and (47.0±16.0)days, respectively ( P>0.05); the number of operation was 3(3, 6) times and 13(4, 17)times, respectively ( P<0.05); the wound healing time was (38.9±17.8)days and (61.8±14.2)days, respectively ( P<0.05). Conclusions:Enterobacteriaceae, Enterococcus and Staphylococcus haemolyticus are the most common pathogenic bacteria for FG, among which the proportion of drug-resistant bacteria is relatively high. NPWT is an effective adjuvant therapy for wound management with reduced operation times and short wound healing time compared to conrentional method.

Objective:To investigate the feasibility of negative-pressure wound therapy (NPWT) in the treatment of uncomplicated cardiac pacemaker pocket infection.Methods:From January 2013 to March 2020, 35 patients with uncomplicated cardiac pacemaker pocket infection were admitted to the Department of Cardiology of Peking University First Hospital, including 21 males and 14 females, aged 27 to 84 years. The retrospective cohort study was conducted. After a complete debridement followed by continuous NPWT (with negative pressure of -16.67 kPa), the pulse-generator was inserted into the new pocket between the musculus pectoralis major and pectoralis minor. Pressure drainage tube was put into the old pocket space. NPWT with the same mode was used again for 5 to 7 days after the wound was closed. The removed pocket tissue of patients was observed with hematoxylin-eosin staining. The wound healing on 10 to 12 days after the operation of pacemaker replacement was observed, and the recurrence of infection was observed during 6 to 42 months follow-up after operation.Results:The fibrous sac wall was observed in pocket tissue of the patients, and the tissue was partially covered with stratified epithelium, with many chronic inflammatory cells infiltration. Multinucleated giant cell reaction was observed in the tissue of some patients. Ten to twelve days after the operation of pacemaker replacement, 35 patients had good wound healing, and sutures were removed. After 6 to 42 months follow-up after operation, 31 patients were cured with no recurrence of infection and the wounds were well-healed; 4 patients who had recurrent infection received whole system of pacemaker removal after the operation.Conclusions:On the premise of complete debridement, NPWT is an alternative treatment for patients with uncomplicated cardiac pacemaker pocket infection.

ObjectiveThe traditional detection method of recombinase polymerase amplificaton (RPA) is not suitable for rapid field detection due to the complicated operation and other factors. Taking the detection of hepatitis b virus (HBV) nucleic acid as an example, it established a detection method of HBV nucleic acid isothermal amplification based on recombinase polymerase amplificaton (RPA) and designed a matching visual detection device of RPA product.MethodsFirstly, a RPA product detection device was designed, which can be used to collect images by taking photos of mobile phones and visually interpret the detection results. Secondly, RPA primers and probes were designed according to the design of HBV gene conserved sequence. Amplification efficiency of each primer pairs were compared though monitoring the RPA reaction of real-time fluorescence curve to screen the best primers and optimize the optimal reaction conditions. Visual detection sensitivity was investigated by using artificial synthesis of HBV target plasmid, and was investigated the specificity of the method by the detection of synthetic plasmid containing hepatitis c virus (HCV), human immunodeficiency virus, treponema pallidum, influenza virus, human papilloma virus DNA fragment. Thirdly, the feasibility of RPA product visualization detection device was verified by comparing with the real-time fluorescence amplification curve. Finally, RPA visual detection was performed on 20 serum DNA samples detected by real-time fluorescence PCR to evaluate the applicability of this method to the detection of actual clinical samples.ResultsThe visual detection device of RPA product was used to realize the negative and positive signals that could be detected by mobile phone photography and visual observation. The visual detection method of HBV nucleic acid RPA amplification could realize the visual detection of DNA targets as low as 1-10 copies of HBV within 30 min at 39 ℃ and had good specificity. The test results of 20 serum DNA samples were completely consistent with those of the commercially available qPCR kit, which preliminarily verified the practicability of the method and the device.ConclusionCombined the established HBV-RPA amplification system with the RPA product visualized detection device, it would be expected to develop a low-cost rapid visualization screening technology platform for HBV nucleic acid in blood.

Objective To evaluate the clinical value of holographic image navigation in urological laparoscopic and robotic surgery.Methods The data of patients were reviewed retrospectively for whom accepted holographic image navigation laparoscopic and robotic surgery from Jan.2019 to Dec.2019 in Beijing United Family Hospital and other 18 medical centers,including 78 cases of renal tumor,2 cases of bladder cancer,2 cases of adrenal gland tumor,1 cases of renal cyst,1 case of prostate cancer,1 case of sweat gland carcinoma with lymph node metastasis,1 case of pelvic metastasis after radical cystectomy.All the patients underwent operations.In the laparoscopic surgery group,there were 27 cases of partial nephrectomy,1 case of radical prostatectomy,2 cases of radical cystectomy and 2 cases of adrenalectomy.In the da Vinci robotic surgery group of 54 cases,there were 51 cases of partial nephrectomy,1 case of retroperitoneal lymph node dissection,1 case of retroperitoneal bilateral renal cyst deroofing and 1 case of resection of pelvic metastasis.There were 41 partial nephrectomy patients with available clinical data for statistic,with a median age of 53.5 years (range 24-76),including 26 males and 15 females.The median R.E.N.A.L score was 7.8 (range 4-11).Before the operation,the engineers established the holographic image based on the contrast CT images and reports.The surgeon applied the holographic image for preoperative planning.During the operation,the navigation was achieved by real time fusing holographic images with the laparoscopic surgery images in the screen.Results All the procedures had been complete uneventfully.The holographic images helped surgeon in understanding the visual three-dimension structure and relation of vessels supplying tumor or resection tissue,lymph nodes and nerves.By manipulating the holographic images extracorporeally,the fused image guide surgeons about location vessel,lymph node and other important structure and then facilitate the delicate dissection.For the 41 cases with available clinical data including 23 cases of robotic-assisted partial nephrectomy and 18 cases of laparoscopic nephrectomy,the median operation time was 140 (range 50-225) min,the median warm ischemia time was 23 (range 14-60) min,the median blood loss was 80(range 5-1 200) ml.In the robotic surgery group,the median operation time was 140 (range 50-215)min,the median warm i schemia time was 21 (range 17-40)min,the median blood loss was 150(range 30-1 200)ml.In the laparoscopic surgery group,the median operation time was 160(range 80-225)min,the median warm ischemia time was 25 (range 14-60)min,the median blood loss was 50 (range 5-1 200) ml.All the patients had no adjacent organ injury during operation.There were 2 cases with Clavien Ⅱ complications.One required transfusion and the other one suffered hematoma post-operation.However,the tumors were located in the renal hilus for these 2 cases and the R.E.N.A.L scores were both 11.Conclusions Holographic image navigation can help location and recognize important anatomic structures during the surgical procedures..This technique will reduce the tissue injury,decrease the complications and improve the success rate of surgery.