Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.
Male ; Mice ; Animals ; Keratin-18 ; Actins ; Desmin ; Liver ; Hepatocytes ; Hepatic Stellate Cells

Objective: To analyze the follow-up and clinical effect of multidisciplinary treatment on the children with spinal muscular atrophy (SMA). Methods: The clinical data including nutritional status, respiratory function, bone health and motor function of 45 children with SMA who received multidisciplinary management 1-year follow-up in the Children's Hospital, Zhejiang University School of Medicine from July 2019 to October 2021 were retrospectively collected. Comparisons before and after management were performed using paired-samples t-test or Wilcoxon rank-sum test, etc. Results: The age of 45 patients (25 boys and 20 girls) was 50.4 (33.6, 84.0) months at the enrollment, with 6 cases of type 1, 22 cases of type 2, and 17 cases of type 3 respectively. After the multidisciplinary management, the cases of SMA patients with malnutrition decreased from 22 to 12 (P=0.030), the level of vitamin D were significantly increased ((45±17) vs. (48±14) nmol/L, t=-4.13, P<0.001). There was no significant difference in the forced vital capacity %pred, the forced expiratory volume at 1 second %pred, and the peak expiratory flow %pred ((76±19)% and (76±21)%, (81±18)% and (79±18)%, (81±21)% and (78±17)%; t=-0.24, 1.36, 1.21; all P>0.05). The Cobbs angle of scoliosis also improved significantly (8.0°(0°, 13.0°) vs. 10.0°(0°, 18.5°), Z=-3.01, P=0.003). The Hammersmith functional motor scale expanded scores of children with SMA type 2 and type 3 both showed significant elevation (11.0 (8.0, 18.0) vs. 11.0 (5.0, 18.5) scores, 44.0 (36.5, 53.0) vs. 44.0 (34.0, 51.5) scores, Z=2.44, 3.11, P=0.015, 0.002). Conclusion: Multidisciplinary management is beneficial for delaying the progression of the multi-system impairments of SMA patients, such as malnutrition, restrictive ventilation dysfunction and scoliosis.
Child ; Male ; Female ; Humans ; Child, Preschool ; Scoliosis ; Retrospective Studies ; Follow-Up Studies ; Muscular Atrophy, Spinal ; Malnutrition

OBJECTIVE: From the perspective of clinical application to analyze the effectiveness and reliability of CPC/PMMA bone cement in percutaneous kyphoplasty (PKP) for the treatment of elderly patients with osteoporotic thoracolumbar fractures. METHODS: A retrospective analysis was performed on 62 patients with osteoporotic compression fracture of single-vertebral thoracic or lumbar segment who underwent PKP surgery and had a bone density less than or equal to -3.0 SD from February 2016 to December 2016. Among them, 23 patients were in CPC/PMMA group, with an average age of (77.6±2.2) years old, 39 patients in PMMA group, with an average age of (77.1±1.1) years old. The indexes between two groups were compared, including the visual analogue scale (VAS), height ratio of anterior vertebra (AVHR), local Cobb angle, cement leakage, new adjacent vertebral fracture(NAVF). RESULTS: There were no significant difference in gender, age, follow-up time and preoperative VAS, AVHR, local Cobb angle between two groups (>0.05), at the 1 day after operation, VAS, AVHR, local Cobb angle in all patients got obvious improvement (<0.05), which was no significant difference at 1 day after operation and final follow-up (>0.05). At the same time, there was no statistically significant difference in the incidence of new adjacent vertebral fracture and cement leakage (>0.05). The pain in both groups continued to improve at follow up after operation (<0.05), the local Cobb angle increased (<0.05) and AVHR decreased slightly (<0.05). However, the images of conventional methods (X-ray or CT) could not find signs about CPC degeneration and new bone ingrowth. CONCLUSION CPC/PMMA composite bone cement is safe and reliablein PKP for treatment of elderly patients with osteoporotic thoracolumbar fractures, which can effectively relieve pain and maintain vertebral body stability. It has the same curative effect as PMMA bone cement. It was worthy to research more in future, although no direct evidences support the CPC/PMMA composite bone cement can reduce the incidence of adjacent vertebral fracture, CPC degeneration or new bone ingrowth.
Aged ; Bone Cements ; Dinucleoside Phosphates ; Fractures, Compression ; Humans ; Kyphoplasty ; Osteoporotic Fractures ; Polymethyl Methacrylate ; Reproducibility of Results ; Retrospective Studies ; Spinal Fractures ; Treatment Outcome ; Vertebroplasty

OBJECTIVE: By comparing the clinical efficacy of short-segment and long-segment fixation for single-segment thoracic and lumbar spine III stage Kümmell disease to explore a more suitable fixed segment for the disease. METHODS: The clinical data of 46 patients with single-segment thoracic and lumbar spine III stage Kümmell disease treated from July 2013 to December 2016 were retrospectively analyzed. Forty-six patients were divided into short-segment fixation group(one vertebra above and below the diseased vertebra) and long-segment fixation group(two vertebrae on the upper and lower of the diseased vertebra) according to different methods of cement stick fixation. There were 25 patients in the short-segment fixation group, including 9 males and 16 females, with an average age of (75.3±4.5) years old, lumbar spine bone mineral density T-value of (-3.1±0.3) g/cm³, follow-up time of (13.0±2.3) months; there were 21 patients in long-segment fixation group, 6 males and 15 females, with an average age of (74.5±3.9) years old, lumbar spine bone mineral density T-value of (-3.2±0.3) g/cm³, follow-up time of (14.7±3.6) months.The gender, age, follow-up time, operation time, intraoperative blood loss, cement leakage, and the rate of adjacent vertebrae fractures were compared between two groups, as well as pain VAS score, ODI, and kyphosis angle before and after surgery. RESULTS: There were no significant differences in age, gender, bone density, pain VAS score, ODI, and kyphosis between two groups before surgery. The operation time and intraoperative blood loss of short-segment fixation group were less than that of long-segment fixation group. The pain VAS score, ODI and kyphosis of the two groups were significantly improved at 7 days after the operation and at the latest follow-up, there was no significant difference between two groups. There were no significant differences in bone cement leakage(9/25 vs 11/21) and adjacent vertebrae fractures(4/25 vs 3/21). CONCLUSIONS Both long-segment fixation and short-segment fixation can effectively relieve pain, correct kyphosis, improve functional index, and achieve better clinical results, but short-segment fixation has less operation time and less intraoperative blood. So single-segment thoracic and lumbar spine III stage Kümmell disease does not need to extend the fixed segment, short-segment fixation is more in line with clinical needs and worthy of further study.
Aged ; Female ; Fracture Fixation, Internal ; Humans ; Kyphosis ; Lumbar Vertebrae ; Male ; Retrospective Studies ; Spinal Fractures ; Thoracic Vertebrae ; Treatment Outcome

OBJECTIVE: To understand the general awareness of the residents around a newly built nuclear power,to evaluate the safety and the attitude on the construction of nuclear power. METHODS: By the stratified random sampling method,1 029 general publics and 260 representatives were selected for investigation. The participants were from 30 km range around the nuclear power plant,each 5 km was assigned as 1 layer and there were total 6 layers. RESULTS: 74. 9%( 965 /1 289) interviewees were not aware of nuclear power,3. 1%( 40 /1 289) interviewees were well or very-well understand nuclear power. The awareness score of the general publics was lower than that of the representatives( 1. 0 vs3. 0,P < 0. 01); male,younger people,higher educational level,higher family income,religious people and living close to the nuclear power plant had higher awareness( P < 0. 05). A total of 28. 4%( 366 /1 289) interviewees considered the nuclear power was very safe or safe. The evaluation of nuclear power safety of the general publics was inferior to the representatives( P < 0. 05). The representatives,older people and those living far away from the nuclear power have better evaluation on the nuclear power safety than the general publics( P < 0. 05). Only 13. 0%( 167 /1 289) interviewees supported building the nuclear power in their residence construction; male,people with family members working in nuclear power plant,people with high degree of awareness on nuclear power,considered highly safety of the nuclear power had higher support for the local nuclear power construction( P < 0. 05). The interviewees who mostly believe about nuclear energy sources were experts,government officials,plant management and domestic media. CONCLUSION: Residents around the nuclear power plant had low awareness of the knowledge about nuclear power. Therefore,it is necessary to enhance the awareness of residents on nuclear power and the trust on the government.

OBJECTIVETo evaluate the efficacy and safty of photoselective vaporization of prostate (PVP) in the treatment of benign prostatic hyperplasia with obstruction within 5 years.

METHODSFrom December 2004 to December 2009, there were 782 cases have been except for neurogenic bladder dysfunction and prostate cancer, who received PVP surgical treatment of BPH. The surgical conditions and postoperative follow-up data were recorded and the follow-up cut-off time for surgery after 5 years.

RESULTSA total of 782 patients with BPH who underwent PVP were included in this retrospective study. The operation in 740 cases was successfully completed at one time. But in other 42 cases, the twice operation was performed. The mean operation time was (85 ± 38) minutes, and the mean energy delivery was (355 ± 124) kJ. The mean catheterization and postoperative hospitalization time was (2.3 ± 1.7) days and (5.2 ± 2.6) days, respectively. No severe intraoperative complications were observed. The mean follow-up was (44.1 ± 19.3) months. The shortest follow-up was 6 months. The longest follow-up was 5 years. Complete follow-up data were available for 398 of the 782 patients. Of the 398 patients followed up for 5 years, the mean international prostate symptom score after 5 years was 12.8 ± 6.9, quality of life score was 2.2 ± 1.6, maximal flow rate was (14.5 ± 2.4) ml/s, and residual urine volume was 58 ml (M50). The retreatment rate because of BPH was 2.3% (9/398). Urethral stricture and bladder neck contracture were observed in 1.5% and 0.5% of the patients, respectively.

CONCLUSIONSPVP has demonstrated remarkably consistent results for objective and subjective voiding parameters. Its late complication is rare and retreatment rate is low.

Aged ; Follow-Up Studies ; Humans ; Lasers, Solid-State ; Male ; Prostatic Hyperplasia ; surgery ; Retrospective Studies ; Transurethral Resection of Prostate ; Treatment Outcome

Objective To study the genotype distribution of extended-spectrum β-lactamases (ESBLs) in ESBLs-producing Escherichia coli (E.coli) isolates from posthepatitic cirrhosis' patients with bloodstream infection.Methods E.coli were isolated in bloodstream from patients with posthepatitic cirrhosis between January and December in 2011.The strains were identified by VITEK-Ⅱ.The antibiol susceptibility tests were performed with K-B method.β-lactamases genes were detected multi-PCR,PCR,sequence and blast.Results A total of 79 non-duplicate clinical isolates of E coli were consecutively collected from liver cirrhosis' patients with bloodstream infection.There were 20 isolates produced TEM-1 type β-lactamases and 1 isolate produced SHV-1 typeβ-lactamases.40 clinical isolates were detected to produce CTX-M type ESBLs,there were 20 CTX-M-1 group and 26 CTX-M-9 group,including 6 stains habouring both CTX-M-1 and CTX-M-9 group.Eight CTX-M genotypes were confirmed by sequencing of the PCR products,including CTX-M-3,CTX-M-14,CTX-M-15,CTX-M-24,CTX-M-28,CTX-M-31,CTX-M-65 and CTX-M-79.Conclusion CTX-M genotype ESBLs was the most popular extended-spectrum β-lactamases in E.coli isolated from liver cirrhosis' patients with bloodstream infection.The CTX-M-14 is the dominant epidemic type.

OBJECTIVEThe effects of sleep deprivation on the immature brain remain unknown. Based on a computer controlled chronic sleep deprivation animal model, the effects of chronic partial sleep deprivation on growth, learning and memory in young rats were explored.

METHODSTwelve weaned male Spraque-Dawley rats (3-week-old) were randomly divided into sleep deprivation, test control and blank control groups. Sleep deprivation was performed using computer-controlled "disc-over-water" technique at 8-11 am daily, for 14 days. The temperature and weights were measured every 7 days. Morris water maze was used to test spatial learning and memory abilities before and 7 and 14 days after sleep deprivation. After 14 days of sleep deprivation, the rats were sacrificed for weighting their major organs.

RESULTSAfter 14 days of sleep deprivation, the rats' temperature increased significantly. During the sleep deprivation, the rate of weight gain in the sleep deprivation group was much slower than that in the test control and blank control groups. The thymus of the rats subjected to sleep deprivation was much lighter than that of the blank control group. After 7 days of sleep deprivation, the rats showed slower acquisition of reference memory, but were capable of successfully performing the task by repeated exposure to the test. Such impairment of reference memory was not seen 14 days after sleep deprivation.

CONCLUSIONSChronic sleep deprivation can affect growth of immature rats, as well as their abilities to acquire spatial reference memory.

Animals ; Body Temperature ; Body Weight ; Growth ; Learning ; Male ; Memory ; Organ Size ; Rats ; Rats, Sprague-Dawley ; Sleep Deprivation ; physiopathology ; psychology

OBJECTIVETo study the anatomy of peroneal tendofascial flap combined with adipofascial flap for the repair of heel tissue defects.

METHODSThe lower extremities of five cadavers (10 sides) were perfused with red latex, the blood supply of peroneal tendofascial flap and vicinity adipofascial flap were observed. The diameter, course, branches and location of the blood vessels were measured. Eight fresh cadavers (16 sides) were perfused with lead oxide-gelatine mixture. The covering fascia tissues of the lower extremities was obtained and photographed by X-ray. The vascular anastomosis and association of nutrient vessel of peroneal tendofascial flap and vicinity adipofascial flap were observed. Two adult lower extremities specimens (4 sides) were used to construct vessel diagrams for observation of the course, distribution and anastomosis of the vessels. Eight cases were treated successfully with theses flaps.

RESULTSThe blood supply of the combined fascial flap is multi-originated. For the area within 4 cm below and above the lateral malleolus cusp, the blood supply includes 2-5 branches from heel lateral artery with an average diameter of (0.5 +/- 0.2) mm, 1-2 branches from posterior lateral malleolus artery with an average diameter of (0.6 +/- 0.2) mm and 2-3 branches from the descending part of perforating branches of peroneal artery with an average diameter of (0.5 +/- 0.2) mm. The blood supply of area 4 cm above lateral malleolus cusp is 1-3 branches from intermuscular septum perforating branches of peroneal artery with an average diameter of (1.0 +/- 0.2) mm. These above branches are anastomosed each other and also send off many smaller branches to form vascular net around tendon. The fascial flaps and free skin grafts in eight patients were completely survived. All patients were followed up for 3-24 months, the donor and recipient sites were healed very well. The functional and cosmetic results were satisfactory.

CONCLUSIONSPeroneal tendofascial flap combined with adipofascial flap, with proximal pedicle or reverse distal pedicle, can be used to repair the defect at the lower leg and refractory small- and medium-sized defects at the heel.

Adipose Tissue ; anatomy & histology ; transplantation ; Adult ; Aged ; Fascia ; anatomy & histology ; transplantation ; Female ; Fibula ; Heel ; injuries ; surgery ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; anatomy & histology ; transplantation ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; Surgical Flaps