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Amyotrophic lateral sclerosis (ALS) is one of the fatal neurodegenerative diseases. Muscle ultrasound can be used in ALS to make early diagnosis, strengthen disease management and differentiate other neuromuscular diseases from it. In ALS patients, morphological changes such as muscle atrophy, increased echo intensity and fasciculation can be detected by muscle ultrasound which is helpful in assessing respiratory and swallowing functions as well. High frequency ultrasound has the clinical value in the diagnosis, monitoring and prognosis evaluation of ALS patients.
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Objective:To investigate the clinical and genetic characteristics of cerebellar ataxia, neuropathy, vestibular areflexia syndrome (CANVAS) with replication factor C subunit 1 (RFC1) gene mutation to improve the understanding of this disease.Methods:A case of CANVAS diagnosed in the Peking University Third Hospital in January 2021 was reported. Detailed genetic analyses of ataxia were performed with DNA extracted from the peripheral blood of the patient. Studies including pathogenic variants of RFC1 gene causing CANVAS were reviewed and the clinical and genetic characteristics of the disease were summarized.Results:The patient was a 51-year-old female with the prominent manifestation of progressive walking instability. And the clinical data met the diagnostic criteria of CANVAS. The genetic tests excluded other hereditary ataxia mutations and identified the biallelic expansion of the pathogenic variant structure (AAGGG)exp repeat amplification in RFC1 gene. A total of 14 studies on CANVAS with RFC1 gene mutation were reviewed. The overall mutation rate of RFC1 gene in CANVAS was 68%-100%, and it varied in sporadic and familial CANVAS. And the mutation had ethnic differences.Conclusions:Among adult patients with late-onset ataxia, the combination of brain magnetic resonance imaging, electrophysiology tests and vestibular function examination is beneficial to the identification of CANVAS. And the genetic test of RFC1 gene has significant value in the diagnosis of this disease. This patient with CANVAS expands the disease spectrum of ataxia in China, and confirms that RFC1 gene mutation is of great significance in the screening of ataxia disorders in the Chinese population.
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Objective:To discuss the surgical techniques and evaluate the clinical effects of laparoscopic varicocelectomy with testicular artery preservation.Methods:In this retrospective study, we collected clinical data of 97 patients with varicocele who underwent laparoscopic varicocelectomy from January 2015 to June 2020. All operations were performed by the same experienced urologist. Conventional laparoscopic varicocelectomy without testicular artery preservation was performed in earlier 35 patients (January 2015 to December 2016), which were taken as control group. The latter 62 patients (January 2017 to June 2020) underwent laparoscopic varicocelectomy with testicular artery preservation were taken as observational group. In observational group, average age was (21.9±6.7)years, there were 47 cases on the left side, 3 cases on the right side and 12 cases bilaterally, totaling 74 sides. There were 22 sides of varicose veinsⅠ, 28 sides of varicose veinsⅡand 24 sides of varicose veins Ⅲ. Clinical manifestations such as scrotal discomfort, pain and scrotal vein masses were observed on 35 sides (47.3%), infertility was observed in 24 cases (38.7%). Average sperm density was (23.7±5.9)×10 6/ml, average sperm motility (grade a+ b) was (33.9±4.1)%. In control group, average age was (23.7±4.6) years, there were 26 cases on the left side, 2 cases on the right side, 7 cases bilaterally, totaling 42 sides. There were 10 sides of varicose veinsⅠ, 17 sides of varicose veinsⅡ, 15 sides of varicose veinsⅢ. Clinical manifestations of scrotal discomfort, pain and scrotal vein masses were observed on 19 sides (45.2%), infertility was observed in 14 cases (40.0%). Average sperm density was (22.3±6.2)×10 6/ml, average sperm motility (grade a+ b) was (32.6±4.8)%.There was no significant statistical difference in preoperative clinical data between two groups ( P>0.05). The observational group followed the procedural steps of freeing the spermatic cord, isolating the testicular artery, and ligating the spermatic vein. The testicular artery was separated by the separating forceps and the electric hook, with the separating forceps holding the spermatic cord fascia in place and the electric hook (without electricity) bluntly separating the blood vessels and lymphatic vessels in the spermatic cord. The operative time, complications, recurrence rate, improvement rate of scrotal symptoms and semen quality, spontaneous pregnancy rate of spouses within 2 years in infertile patients were compared between the two groups. Results:The mean operative time in observational group was longer than control group [(35.8±7.7)min vs.(16.5±5.5)min, P<0.001]. Occurrence of postoperative acute epididymitis was lower in observational group compared to control group [1.4% (1 side) vs. 11.9% (5 sides), P<0.05] . No testicular atrophy (0 side) occurred in observational group, however, this complication could be found in 7.1% (3 sides) of control group ( P<0.05). Improvement rate of scrotal symptoms and semen quality was higher in observational group than that in control group after operations [77.1% (27 sides) vs. 47.4% (9 sides), P<0.05; and 72.6% (45 cases) vs.51.4% (18 cases), P<0.05]. The rates of spousal natural pregnancy within 2 years in infertile patients was higher in observational group than that in control group [70.8% (17 cases) vs. 50.0% (7 cases), P<0.05]. The rates of hydrocele and scrotal edema were similar in two groups [9.5% (7 sides) vs. 9.5 (4 sides)%, P>0.05], and the recurrence rate of varicocele was similar [8.1% (6 sides) vs. 7.1% (3 sides), P>0.05), without statistically significant difference. Conclusions:Using separating forcep and electronic hook can help to separate the testicular artery when performing laparoscopic varicocelectomy. In this operation, to preserve the testicular artery can get better effects an less complications.
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Objective:To construct a Nomogram model in predicting recurrence-free survival (RFS) and overall survival (OS) at six months, one year and two years after hepatocellular carcinoma (HCC) resection by using inflammatory markers combined with other routine clinical indicators.Methods:The data of 314 patients with HCC who underwent first time hepatectomy at Beijing Chaoyang Emergency Rescue Center and Air Force Characteristic Medical Center from January 2013 to January 2018 were analyzed. HCC patients who underwent hepatectomy at the First Medical Center of PLA General Hospital from January 2011 to January 2016 ( n=106) were used as the external validation group. Univariate and multivariate Cox proportional risk model was used to analyze independent risk factors of recurrence and death in HCC patients. A Nomogram model was constructed based on independent risk factors. Validation of the efficacy of the Nomogram model was done based on external data. Results:In the experimental group, 174 patients relapsed. The median RFS was 26 months. The 6 months, 1 year and 2 years RFS were 26.8%, 43.9%, and 68.8%, respectively. A total of 142 patients had died. The median survival time was 30 months. The 6 months, 1 year and 2 years OS were 5.9%, 23.6% and 63.1%, respectively. In the external validation group, 63 patients had developed recurrence, with a median RFS time of 28 months. The 6 months, 1 year and 2 years RFS were 26.4%, 45.3%, 54.7%, respectively. The median survival time was 31 months. The 6 months, 1 year and 2 years OS were 7.5%, 25.5%, 46.6%, respectively. Tumor size (>6.0 cm, HR: 1.447), vascular invasion ( HR: 1.408), TBil (>0.94 mg/dl, HR: 1.949), NLR (>2.54, HR: 2.843), AGR (≤0.88, HR: 2.447) were independent risk factors of HCC recurrence ( P<0.05). Tumor size (>6.0 cm, HR: 2.207), vascular invasion ( HR: 1.529), and NLR (>2.54, HR: 2.708) were independent risk factors of death for HCC patients ( P<0.05). The C-indexes of half-year, one-year and two-year RFS were 0.764 (95% CI: 0.677-0.854), 0.710 (95% CI: 0.615-0.824) and 0.673 (95% CI: 0.601-0.786), respectively. The C-indexes of half-year OS, one-year OS and two-year OS were 0.729 (95% CI: 0.648-0.841), 0.708 (95% CI: 0.608-0.813) and 0.664 (95% CI: 0.618-0.771), respectively. Conclusion:In this study, the construction of a Nomogram model in predicting prognosis of HCC patients was helpful to guide clinicians in improving preoperative treatment plans and in providing ideas for individualized treatment of patients.
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Objective To study the curative efficacy and safety of single-unit umbilical cord blood transplantation (sUCBT) for malignant hematologic diseases,which is provided by China's public cord blood bank.Methods We retrospectively analyzed 409 cases of malignant hematologic diseases who accepted myeloablative single-unit unrelated donor UCBT without ATG at our center between May 2008 and December 2016.A comparative analysis was made on the total nuclear cells (TNC) of the umbilical cord blood before freezing and after thawing,the cells of CD34+,the recovery rate of cells and the clinical effect of UCBT.Result 409 units of umbilical cord blood used in UCBT respectively came from eight China's public cord blood banks.The average TNC of 409 units of umbilical cord blood before freezing and after the tubular recovery were respectively 18.5 × 108 and 16.34 × 108 (p =0.000).The average recovery rate of the tubular recovery was 88.5％,and there was significant difference among cord blood banks (P =0.000).The average TNC of umbilical cord blood before freezing and transfusion were respectively 18.5 × 108 and 15.86 × 108 (p =0.000).The average recovery rate of umbilical cord blood transfusion was 85.9％,with the difference being significant among cord blood banks (P =0.000).The average number of CD34+ cells before freezing and after the tubular recovery was 11.18 × 106and 8.68 × 106 (p =0.000).The average recovery rate of CD34+ cells after the tubular recovery was 80.75 ％,with the difference being significant among the cord blood banks (P =0.000).At 42nd day after UCBT,the cumulative incidence of neutrophil engraftment was 95.4％,and the median time of the engraftment was 17 days (11-38 days).The cumulative incidence of platelet engraftment at 120th day was 84.6％,and the median time of the engraftment was 36 days (14-93 days).The cumulative incidence of erythrocyte engraftment at 60th day was 92％,and the median time of engraftment was 22 days (9d-60 days).After the umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of engraftment.The P values for cumulative incidence of neutrophil,platelet and erythrocyte engraftment were respectively 0.004,0.01 and 0.000 2,with the differences being statistically significant.At 100th day after UCBT,the cumulative incidence of Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees of acute graft-versus-host disease (aGVHD) was respectively 28.63％ and 15.7％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of aGVHD.There was no significant difference between Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees (P =0.809 and 0.68 respectively).At 3rd year after UCBT,the cumulative incidence of relapse was 15.89％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the cumulative incidence of relapse (P =0.898).At 3rd year after UCBT,the overall survival (OS) rate and disease free survival (DFS) rate were respectively 66.7％ and 59％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in OS and DFS.There was no significant difference in OS and DFS (P =0.566 and 0.703 respectively).At 3rd year after sUCBT,the rate of graft-versus-host diseases/relapse-free survival (GRFS) was 54.3％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the rate of GRFS (P =0.449).Conclusion The umbilical cord blood provided by China's public cord blood bank was used in UCBT.It has a high safety and good efficacy in treating malignant hematologic diseases.But it needs to set up the standardized and normalized quality-control system of umbilical cord blood for China's public cord blood bank.
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Objective To establish a cultivating method for obtaining a large number of P0 generation human placental chorionic-derived mesenchymal stem cells (hpcMSCs).Methods The hpcMSCs were isolated from human placental chorion.After primary culturing and culturing for seven days,the culture medium,the non-adherent tissue and the douching normal saline of the primary culture were centrifuged and re-cultured twice.Cell morphology was observed by an inverted microscope.CCK-8 was used to measure the cell growth curve.Flow cytometry was used to detect cell surface markers.Adipogenic and osteogenic differentiation kits were used to assess the cell differentiation potential.Results The obtained hpcMSCs were fibroblast-like adherent cells and (25.54±3.38)×106 cells were obtained per placenta.The total yield of the primary culture,secondary culture and tertiary culture were (11.73±2.09)×106,(11.12±1.42)×106 and (2.69±0.71)×106,respectively,and the incubation time were (12.00±0.64) d,(8.87±0.63) d and (12.33±0.80) d.There was significant differences in incubation time between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the incubation time of the tertiary culture had an increasing trend (P＞0.05).The yield per culture flask of the primary culture,secondary culture and tertiary culture were (1.12±0.15) × 106,(2.10±0.16)×106 and (1.04±0.16)×106,respectively.There was significant differences in the yield per culture flask between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the yield per culture flask of the tertiary culture had a decreasing trend (P＞0.05).There was no difference among the three cultures in the growth curve and the expression of surface markers,and the osteogenic and adipogenic differentiation were all positive.Conclusions The P0 generation hpcMSCs isolated from a choriocarcinoma sample can be doubled by the three cultures compared with the primary culture,which can provide plenty stem cell source for the regenerative medicine.
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BACKGROUND:There are a lot of studies on isolation and culture methods of human placental chorionic-derived mesenchymal stem cells (hpcMSCs), but how to simply and efficiently harvest a large amount of primary MSCs has not been resolved. OBJECTIVE:To optimize the tissue explants method of isolating and culturing hpcMSCsin vitro. METHODS:Human placental chorionic villi were collected from full-term deliveries under aseptic condition and isolated by electric homogenizer. hpcMSCs were prepared by tissue explants method. The fluid and tissue of the primary culture flask and douching normal saline of the initial culture were centrifuged and prepared for secondary culture. RESULTS AND CONCLUSION: It saved time and effort to treat human placental chorionic villi with electric homogenizer, with good effects on tissue dispersion and removal of red blood cells. The average time of cell acquisition in initial culture and secondary culture was (17.73±1.14) and (10.03±1.30) days, respectively. The yields of primary cultured cells in initial culture and secondary culture were (6.97±0.98)×105 and (13.82±1.44)×105per Φ100 mm culture dish, respectively. The adherent cells showed fibroblast-cell-like shape, which were in parallel or circinate arrangement. Highly expressed CD73, CD105 and CD90 could be detected in the third generation of hpcMSCs, but CD34, CD45, CD14, CD19 and HLA-DR were negative. Following induction, alizarin red staining and oil red O staining produced a strong reaction in cells. In a word, the optimized method is a simple and efficient method for obtaining a large amount of primary hpcMSCs.
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Objective To evaluate the clinical significance of direct trocar insertion using optical trocar in the establishment of the primary port during trans-peritoneal laparoscopic surgical procedures.Methods A prospective study was conducted by collecting the data of 120 patients who should be performed abdominal laparoscopic surgery from April 2015 to December 2015.The 120 patients were randomly divided into a research group and a control group.The research group consisted of 34 male patients and 26 female patients,mean age was (52.0 ± 11.9) years and mean BMI was (24.9 ± 2.9) kg/m2.In research group,patients were positioned laterally with the flank padded and elevated.A predetermined position was drawn prior to surgery between the umbilicus and lateral rectus abdominis,for the creation of the primary laparoscopic trocar port.The predetermined point was incised,and then the method of direct trocar insertion using the optical access trocar was used for establishment of the primary port.After this maneuver was completed the surgery continued as indicated.The control group consisted of 36 male patients and 24 female patients,whose mean age was (52.9 ± 11.4) years and mean BMI was (25.2 ± 2.4) kg/m2.This group underwent the traditional method of port construction by incision into the abdomen.The time of constructing the passage,leakage rate,bleeding rate,and injury rate of abdominal organs were compared.Results In research group,the time of building primary port was clearly shorter than that in control group (2.7min vs.15.9min,P ＜ 0.05),the leakage rate was also obviously reduced compared to that in control group (0 vs.30％,P ＜ 0.05).Neither groups observed any significant bleeding nor visceral organ damage throughout the study.Conclusion Direct trocar insertion using optical trocar to establish observation port is a highly efficient and safe method in trans-peritoneal laparoscopic operation,which should be research thoroughly in clinical practice.
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Objective To verify whether the storage bag can reach the use requirement of cord blood freezing storage by conduc-ting a series of tests before staring use of new type cryopreservation bag for cord blood.Methods According to the standard opera-tion procedure,28 new type cryopreservation bags were extracted.The cord blood units were performed the cryostorage and thaw for rewarming according to the standard operating procedures.Then the physical integrity and various quality indicators of cord blood in bag were detected,including the nucleated cell viability,cell recovery rate,CD34-positive cell,colony-forming cultivation of stem cells,sterility test and verification of actual load volume.Results All the tested storage bags passed the integrity test,and the various testing indicators of the preserved cells conformed to the use requirements.Conclusion The new type storage bag has the same performance and effect to original bag,it is verified that the maximum loading volume of 50 mL(for 50 mL specification of storage bag)and 80mL(for 250mL specification of storage bag)is safe.
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BACKGROUND:Fetal bovine serum based media used for expanding and cryopreserving human mesenchymal stem cells raise safety concerns in the clinical setting. OBJECTIVE:To investigate the feasibility of human umbilical cord blood plasma as a replacement for fetal bovine serum in culture and cryopreservation of human mesenchymal stem cells derived from umbilical cord. METHODS:Umbilical cord blood units were suitable for this research if they fulfil ed the donor selection criteria of the Guangzhou Cord Blood Bank strictly. Cord blood plasma was ready to use after col ected from the plasma reduction during the suitable cord blood units processing and pooling. Umbilical cord mesenchymal stem cells were harvested from the umbilical cord tissue of health ful-term newborns after delivery by enzyme digestion and were cultured in the presence of Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing either fetal bovine serum or pooled cord blood plasma. Morphology, proliferation, immunophenotype detected by flow cytometry and differentiation toward adipogenic and osteogenic lineages were utilized for investigating the effect of media on umbilical cord mesenchymal stem cells after 3-5 passages. Then cells were cryopreserved in media containing 10%dimethyl sulfoxide, 20%fetal bovine serum or 20%pooled cord blood plasma for at least 6 months. Viability, adhesion, proliferation, immunophenotype and osteogenic differentiation of the cells were assessed after thawing. RESULTS AND CONCLUSION:The morphology (spindle-shaped and plastic-adherent), phenotype and differentiation potential (osteogenic and adipogenic) were almost indistinguishable between cells cultured in fetal bovine serum or cord blood plasma medium, while cells grown in cord blood plasma medium demonstrated significantly higher proliferation rates than those in medium containing fetal bovine serum. After thawing, the cells maintained their adherence to the culture surface and differentiation potential to osteoblasts, but cells from cord blood plasma cryopreservation medium showed significantly better plastic attachment and produced greater cellnumbers than fetal bovine serum for the first three post-thaw passages. The results demonstrate that cord blood plasma can sever as an effective substitute to fetal bovine serum for growth, maintenance and differentiation of umbilical cord mesenchymal stem cells, and thus it wil be a safe choice for clinical-scale production of human mesenchymal stem cells.
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Objective To summarize the common types and clinical characteristics of ureter disease;which can increase manipulation difficulties and adverse events during rigid ureteroscopic procedures. Methods From Jan 2001 to Dec 2010,our team performed 317 rigid ureteroscopic Drocedures for ureteroscopic examination or treatment;including 60 difficult procedures(34 male and 26 female).The mean age of the patients was 37 years (range,18 to 71).The ureteral diseases were classifted into five types according to the pathological characteristics:Type Ⅰ calculous stenosis,Type Ⅱ neoplastic stenosis;Type Ⅲ non-congenital stenosis,Type Ⅳ congenital stenosis,Type Ⅴ expansion of tortuous ureters.The operative time,complications,and conversion to open surgery were evaluated,and the therapeutic methods were analyzed. Results Of the 60 difficuhly-manipulated procedures,the mean manipulated time was 75 min (range,31 to 200).Intra-operative complications occurred in 9 procedures,including 4 cases of mucosal bleeding,2 cases of submucosaI false passage and 3 cases of ureteral perforation.Eleven procedures were converted to open surgery. In five procedures only a double J tube was inserted for drainage due to the difficulty of entering the ureter.Fiftyfive patients were followed up for 17 months (range,3 to 110);48 patients were cured,5 patients improved and 2 patients were unchanged. Conclusions The five types of ureteral disease can increase operative difficulties and risks of rigid ureteroscopic procedures.We should be cautious during surgery and should stop manipulation or convert to other surgeries if necessary.