This study aimed to investigate the therapeutic effects of Morinda officinalis iridoid glycosides (MOIG) on bone loss of rheumatoid arthritis (RA) rats, and the mechanism of osteoclast function and activity induced by lipopolysaccharide (LPS). RA rats were established by injecting bovin type II collagen. The Bio-ethic Committee of Zhejiang Chinese Medical University approved all experimental protocols associated with this study (IACUC-20180410-03). The collagen-induced arthritis (CIA) rats were administered drug by gavage for 8 weeks; the femoral trabecular micro-structure changes were observed in CIA rats by micro-CT; the LPS-induced osteoclasts model further observed the effect and mechanism of anti-inflammatory osteoporosis in vitro. The results indicated that MOIG could markedly increase bone mineral density (BMD) in CIA rats, improve trabecular micro-structure. In vitro studies demonstrated that MOIG could significantly inhibit osteoclastogensis and differentiation, suppress tartrate resistant acid phosphatase (TRAP) activity, F-actin ring formation, TNF receptor associated factor 6 (TRAF6) recruitment, and inhibitor of nuclear factor kappa-Bα (IκBα) degradation as well as p65 phosphorylation, thereby repressing nuclear factor kappa-B (NF-κB) signaling pathway activation. Subsequently, MOIG effectively inhibited osteoclast nuclear factor of activated T-cells c1 (NFATc1) and cellular oncogene Fos (c-Fos) expression, as well as bone resorption related protein activity including matrix metalloprotein 9 (MMP-9) and cathepsin K (CtsK). Meanwhile, MOIG also repressed the phosphorylation expression of Janus activating kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3), thereby inhibiting JAK2/STAT3 signaling pathway activation. Moreover, further studies found that MOIG could suppress glycogen synthase kinase-3β (GSK-3β) activity, and GSK-3β gene silencing could markedly inhibit oetsoclast F-actin ring formation as well as the phosphorylation expression of p65 and STAT3. Of note, compared with GSK-3β gene silencing group, there was no significant difference in the group treated with both MOIG with GSK-3β gene silencing simultaneously. Thus, the results suggested that MOIG may inhibit NF-κB signaling pathway and JAK2/STAT3 signaling pathway activation via regulating GSK-3β, thereby alleviating bone destruction in RA.

Objective To test the performances and applicability of the pressure infusion cuff and blood warmer under severe cold environments in the field.Methods Under the severe cold conditions in the field,the simulated hemorrhagic shock patient was placed on a stretcher and transported to a transfer vehicle 2 km away,a pressure infusion cuff and blood warmer was used to perform fluid resuscitation and heating and pressurization blood transfusion during the transport,and the effect of heating and pressurization of the device during the transport was evaluated;two healthy adult male pigs were selected to construct animal models of hemorrhagic shock from war wounds,of which one pig was used as an experimental animal to be treated by blood transfusion with a pressure infusion cuff and blood warmer and the other pig was recognized as a control animal to be treated by blood transfusion using commercially available inflatable pressurized bags and thermostats,and comparison analysis was carried out on the changes in vital signs of animals before and after treatment by the 2 methods of warming and pressurization and the effects of warming and pressurization transfusion.Results The pressure infusion cuff and blood warmer gained advantages in convenience,efficiency and applicability during patient transport;the experimental animal behaved better than the control animal in vital signs such as blood volume,pressure and respiration,transfusion speed and fluid warming.Conclusion The pressure infusion cuff and blood warmer has high efficiency and is applicable under severe cold conditions in the field.[Chinese Medical Equipment Journal,2023,44(11):117-120]

【Objective】 To learn the production efficient of platelet components among prefecture-level blood stations in China, to provide supporting data for those blood stations to optimize the production mode of platelet components and continuously improve production efficiency and supply capacity. 【Methods】 The data from 2017 to 2020 was obtained from 24 prefecture-level blood stations who were the members of the practice comparison network for blood institutes in China. The collection units of apheresis platelets, the number of dual-collections of apheresis platelets and plasma, the average apheresis units of one platelet apheresis procedure, the discarded rate of apheresis platelets, the amount of expired apheresis platelets and the amount of apheresis platelets issued were collected. For concentrated platelets, the prepared amount of platelet concentrates and the amount of expired platelet concentrates were collected; both the quantity of qualified and issued concentrated platelets were submitted for statistical analysis.The total output and efficiency of platelet components were calculated based on the collected data. 【Results】 The average annual growth rate of apheresis platelets collection in 24 prefecture-level blood stations was 12.23%, accounting for 99.80% of the total platelet output; the average collection unit of one platelets apheresis procedure was 1.75; from 2019 to 2020, only 5 blood stations performed dual-collection of platelet and plasma during one apheresis procedure; the discarded rate of apheresis platelets was 0.28%, of which 0.007% was due to expiration. A total of 1 621.2 therapeutic units of concentrated platelets were prepared, and 13.03% of them was discarded due to the expiration. The production efficiency of platelet components was 97.56%, of which the production efficiency of apheresis platelets was 97.61% and the production efficiency of concentrated platelets was 74.43%. 【Conclusion】 There are large regional differences in the supply capacity of platelet components in prefecture-level blood stations. Apheresis platelets are the main resource of platelet components product, and the collection capacity is increasing over the years with the characteristics of high production efficiency and low expiration scrapping rate. However, the preparation of concentrated platelets are still limited with relatively low production and high expiration discarded rate.

OBJECTIVE: To summarize the clinical outcomes of partial pubectomy assisted anastomotic urethroplasty for male patients with pelvic fracture urethral distraction defect (PFUDD) and discuss the skills of partial pubectomy. METHODS: The clinical data of 63 male patients with PFUDD were retrospective reviewed. The procedure of the anastomotic urethroplasty was as follows: (1) circumferentially mobilizing the bulbar urethra; (2) separating the corporal bodies; (3) performing the urethral anastomosis after partial pubectomy and exposure of the healthy two ends of the urethra. RESULTS: The mean age of the patients was (39.2±15.6) years (range: 15-72 years). The median time between incidents and operation was 15 months (range: 3-240 months) and the mean length of stricture was (3.85±0.91) cm (range: 1.5-5.5 cm). All the patients had undergone suprapubic cystostomy in acute setting. Thirteen patients (20.6%) were re-do cases and the patients who had undergone dilation, direct vision internal urethrotomy (DVIU), and open primary realignment were 22 (34.9%), 8 (12.7%), and 8 (12.7%), respectively. Assisted with partial pubectomy, the anastomotic urethroplasty had been successfully performed in all the patients. The mean time of operation was (160.2±28.1) min (110-210 min), and the mean evaluated blood loss was (261.1±130.3) mL (100-800 mL). There were 3 cases (4.8%) with perioperative blood transfusions. The postoperative complications were bleeding and urinary tract infection, which were controlled conservatively. The mean maximum urine flow rate was (23.7±7.4) mL/s (15.0-48.2 mL/s) after removing the catheters 4 weeks after urethroplasty. The median follow-up was 23 months (12-37 months). The urethroscopy showed 2 cases of stricture recurrences and 1 case of iatrogenic penile urethral stricture due the symptoms of urinary tract infection and decreased urine flow and all of them were successfully managed with dilation. CONCLUSION Partial pubectomy can effectively reduce the gap between the ends of the urethra and promote tension-free anastomosis during the anastomotic urethroplsty for patients with PFUDD. The skills of the procedure include good exposure of the anterior surface of pubic symphysis between the separated corporal bodies, carefully mobilizing and securing the deep dorsal vein of the penis, removing the partial pubic bone and the harden scar beneath the pubic bone for good exposure of the proximal urethral end.
Adolescent ; Adult ; Aged ; Anastomosis, Surgical ; Humans ; Male ; Middle Aged ; Pelvic Bones/surgery* ; Retrospective Studies ; Treatment Outcome ; Urethra/surgery* ; Urethral Stricture/surgery* ; Urologic Surgical Procedures, Male ; Young Adult

OBJECTIVE: To study the association of copy number of SMN1 and SMN2 with clinical phenotypes in children with spinal muscular atrophy (SMA). METHODS: A total of 45 children with SMA were enrolled. Multiplex ligation-dependent probe amplification was used to measure the gene copy numbers of SMN1 and SMN2. The association of copy number of SMN1 and SMN2 with clinical phenotypes was analyzed. RESULTS: Of the 45 children with SMA, 42 (93%) had a homozygous deletion of SMN1 exons 7 and 8, and 3 (7%) had a deletion of SMN1 exon 7 alone. No association was found between SMA clinical types and the deletion types of SMN1 exons 7 and 8 (P>0.05). There was a significant difference in the distribution of SMN2 gene copy numbers between the children with SMA and the healthy children (P<0.05). The children with SMA usually had two or three copies of SMN2 gene, while the healthy children usually had one or two copies of SMN2 gene. There was a significant difference in the distribution of SMN2 copy numbers among the children with different SMA clinical types (P<0.05). The children with two copies of SMN2 gene had a significantly lower age of onset than those with three or four copies. Most of the children with type I SMA had two or three copies of SMN2 gene. Most of the children with type II SMA had three copies of SMN2 gene. Most of the children with type III SMA had three or four copies of SMN2 gene. Children with a higher copy number of SMN2 gene tended to have an older age of onset and better motor function and clinical outcome, and there was a significant association between SMN2 gene copy number and clinical outcome (P<0.05). CONCLUSIONS The SMN2 gene can reduce the severity of SMA via the dosage compensation effect. SMN2 copy number is associated with the phenotype of SMA, and therefore, it can be used to predict disease severity.
Child ; Humans ; Muscular Atrophy, Spinal ; genetics ; Phenotype ; Survival of Motor Neuron 1 Protein ; genetics ; Survival of Motor Neuron 2 Protein ; genetics

OBJECTIVE: To explore the clinical value of non-invasive prenatal testing(NIPT)for screening fetal chromosomal copy number variations(CNVs) and microdeletion/microduplication syndromes(MDs).METHODS: Retrospective analysis was made in the 10 005 women who received NIPT during the first trimester(15-20+ 6 weeks)from January,2012 to July,2017,at First People's Hospital of Yunnan Province,Department of Genetic Diagnosis Center.Among them 32 pregnant women were indicated fetal CNVs,25 of 32 pregnant women selected interventional prenatal diagnosis.Statistical analysis was made on the amniotic fluid/cord blood chromosome G band karyotype and high-throughput sequencing(NGS)genome copy number analysis was made,and relevant CNVs were searched and analyzed in the corresponding database;the consistency of CNVs found in NIPT with interventional prenatal diagnosis was statistically analyzed.RESULTS: During the second trimester(15-20+ 6 weeks),in the 10 005 pregnant women who received NIPT testing 32 cases were shown to have high risks of fetal CNVs,and the screening positive rate was 0.32%(32/10 005).In 25 high risk pregnant women who accepted invasive prenatal diagnosis via informed choice,14 women wereconfirmed as fetal CNVs,the positive predictive value(PPV)of NIPT being 56%(14/25),including 9 cases of microdeletion and 5 cases of microduplication.The sizes were between 587.75 kb and 36.05 Mb.The size and the start and end positions of CNVs found by NIPT were similar to those of fetal DNA samples detected by NGS.Among 14 cases of fetal CNVs,11 cases were identified as MDs,3 cases as unknown clinical significance.In 11 cases of MDs,8 cases were observed fetal chromosome structure abnormalities by karyotype analysis,10 cases were confirmed as de novo abbreviations,and 2 cases as originated from paternal same MD.After genetic counseling,10 pregnant women in 11 cases of MDs chose informed terminations,and one case chose continuing pregnancy.CONCLUSION: As a high-precision screening method,NIPT is expected to be an effective mean to screen for fetal CNVs,which can be used to detect highrisk chromosome microdeletion and microduplication CNVs of larger segments.High risk cases of fetal CNVs found by NIPT require invasive prenatal diagnosis for validation.

Epilepsy clinically has an inhibitory impact on cognitive function, but whether it is associated with epileptogenesis is unclear. Since the epileptic spike characterizes temporal lobe epilepsy (TLE), the present study was aimed to analyze the transient effects of sporadic spikes (SSs) on theta rhythm during epileptogenesis. The local field potentials (LFPs) were recorded in CA1 area in four rats with the pilocarpine injections during exploration, and theta phase stability and power were globally estimated around SSs, also during prolonged period without SS (both as experiments) as well as pre-injections (control). Finally, the LFPs were simulated by changing the average excitatory and inhibitory synaptic gain values (including slow and fast inhibition loops) with the help of simplified dynamical model of CA1 networks, and then theta phase stability was evaluated in several cases. It was found that the SSs could have negative impacts on theta rhythm both transiently and persistently, which may be dependent on the temporal courses leading to epilepsy, being acuter in early stage than later stage, but even in latent stage, theta power was strong. The simulations partly demonstrated that the synaptic imbalance concomitant with the occurrence of SSs might be related to the dynamics of theta phase stability. The results indicate that the SSs might have persistent negative impacts on the cognition rhythm, and the effects might alter during epileptogenesis, leading to the cognitive dysfunction.
Animals ; CA1 Region, Hippocampal ; physiopathology ; Epilepsy, Temporal Lobe ; chemically induced ; physiopathology ; Pilocarpine ; Rats ; Theta Rhythm

BACKGROUNDSurgical treatment of acetabular fracture has long been a challenging area in the field of orthopedic trauma. The aim of this research was to investigate the operative methods for delayed acetabular fractures and to assess the operation results.

METHODSThe operative approaches, procedures, results, and complications of the delayed acetabular fractures between 1995 and 2005 were retrospectively evaluated at Beijing Jishuitan Hospital. Quality of life was assessed for each patient with the Merle d'Aubingne and Postel fracture function rating scale and the radiological result was assessed using the Matta radiological score.

RESULTSSixty-eight cases (70 hips) were followed up with a minimal duration of five years (average of 5.8 years). Excellent functional results were observed in 10 hip joints, good results in 40, fair results in 11, and poor results in nine. The risks of poor prognosis include impact fracture or osteochondral fracture of femoral head, a time beyond 42 days from injury to operative management, and dislocation of femoral head during the injury. Some of the problems, which were observed included postoperative infection in two hips, iatrogenic sciatic nerve injury in eight hips, traumatic arthritis in 15 hips, heterotopic ossification in 17 hips, and necrosis of the femoral head in six hips.

CONCLUSIONA careful selection of operative indications for delayed acetabular fractures in combination with a proper operative approach and appropriate reduction and fixation could guarantee relatively good results.

Acetabulum ; injuries ; surgery ; Adolescent ; Adult ; Female ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo establish an improved three-dimension (3D) and serum-free approach to differentiate human embryonic stem cells (hESCs) into endothelial cells, and detect the endothelial functions of the obtained cells.

METHODSWe cultured undifferentiated H9 human embryonic stem cell line in low-adhesion dishes to form embryonic bodies (EBs). After 12 days, EBs were harvested, re-suspended into rat tail collagen type I, and put into the incubator (37℃). After 30 minutes, EGM-2 culture medium was added to the solidified collagen, and the EBs were cultured for another 3 days to form embryonic body-sproutings (EB-sproutings). EB-sproutings were digested with 0.25% collagenase I and 0.56 U/ml Liberase Blendzyme for 20 minutes respectively, and the CD31(+) cells were sorted by FACS. The endothelial functions were tested by Dil-ac-LDL uptake assay and tube formation assay.

RESULTSThis approach raised the efficiency of endothelial differentiation to 18%, and also avoided the contamination with animal materials. The obtained hESC-derived endothelial cells (hESC-ECs) had the similar pattern of surface biomarkers as human umbilical vein endothelial cells (HUVECs), and their endothelial functions were confirmed by the uptake of Dil-ac-LDL and the tube formation on Matrigel.

CONCLUSIONSThe improved 3D approach can enhance the efficiency of differentiation from hESCs into endothelial cells. Furthermore, serum free differentiation system may be applied in future hESC-based therapies for various ischemic diseases.

Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Line ; Collagen Type I ; Culture Media ; Embryonic Stem Cells ; cytology ; Endothelial Cells ; cytology ; Humans

OBJECTIVETo summarize experience of pediatric intensive care and explore the incidence of complications, the involved pathogens among liver recipients to determine the effective strategies for preventing complications.

METHODSBetween June 2006 and July 2009, 35 children under the age of 14 yr received 35 liver transplantations (LTs) performed at the center. A retrospective review of 22 infants weighing 8.8 kg or less underwent 23 transplants was conducted. Indication for transplantation was biliary atresia. Central venous pressure and arterial blood pressure were monitored continuously and fluid monitoring was performed every 2 hours in the first postoperative week. Blood loss, ascites, and intraoperative transudate loss were primarily replaced with 5% albumin and crystalloids to maintain a central venous pressure between 4 and 6 cm H(2)O. Oral food intake was allowed as soon as possible. To identify vascular or biliary complications, liver doppler ultrasound was performed intraoperatively immediately after reperfusion and after closure of the abdominal wall and postoperatively, twice daily during the first week after surgery. Immunosuppression was initially cyclosporine based, in combination with steroids. Cyclosporine was begun one day prior to transplantation at a dose of 10 mg/(kg·d) divided into two doses, except for cases with hepatic encephalopathy and severe infection. The subsequent doses were adjusted on the basis of recommended trough blood concentrations at different stages. Steroids were eventually discontinued at a time point exceeding 6 months after transplantation. The diagnosis of rejection was confirmed by histology on needle biopsy specimens. Acute graft rejection episodes were treated with a 3-day scheme of IV methylprednisolone 10 mg/(kg·d) followed by recycling doses during the following 3 days (7.5, 5 and 2.5 mg/(kg·d).

RESULTSThe most common postoperative complications were infections (18 cases), gastrointestinal bleeding (3 cases), and vascular complications (4 cases). Rejection occurred in 25% of patients. There was one perioperative death from primary graft non-function. The most common isolated bacteria of the pathogen spectrum were Staphylococcus epidermidis. The median length of stay (LOS) in the PICU for 22 patients (23 transplants) was 10 days (range 5 - 21) and the mean length of stay in the hospital was (18.5 ± 116) days (range, 11 - 48 days). Mean requirement for artificial ventilation was 37.6 h. Mean use of dobutamine, prostaglandin E1 and dopamine was 3.3, 7.5 and 8.8 days, respectively. Preoperatively, 3 children had gastrointestinal bleeding, 18 had ascites, 2 had encephalopathy, 22 had jaundice, and 16 had coagulopathy. There were multiple early operative complications in these infants, including one graft with primary non-function (4.5%). Two patients (9.1%) returned for a total of three times for gastrointestinal bleeding or intra-abdominal hematoma. Three patients (13.6%) had early postoperative intestinal perforations related to adhesions or enterotomy, one was associated with a bowel obstruction. There were 26 episodes of bacterial or fungal infections in 18 (81.8%) patients in the early postoperative period, and infection was the direct/contributing cause of death in one infant. These infections included pneumonia, intra-abdominal abscess or sepsis. All of the bacterial and fungal infections were successfully treated with the appropriate antibacterial and antifungal agents, except for one patient who developed overwhelming sepsis after small bowel perforation. Four (18.2%) patients developed five episodes of acute allograft rejection during the first 15 days after LT. Three of the four patients who developed rejection were transplanted before 2007. All episodes of rejection were treated successfully with intravenous steroid pulse and optimization of cyclosporine levels or FK506 conversion. Of the 20 survivors beyond the perioperative period, two cases (10%) had hypertension requiring therapy.

CONCLUSIONSLiver transplantation in infants with biliary atresia appears technically demanding but acceptable. There should be essentially no age or size restriction for infants and transplantation can be performed with good outcome, although the frequency of complications is much higher than that seen in older children. The improvement in medical and nursing expertise in this group of very sick infants is based on judicious preoperative donor and recipient selection, meticulous surgical technique (vascular reconstruction and abdominal closure), immediate detection and prompt intervention of complications, and keen postoperative surveillance, which reflect a learning curve for both the technical aspects of liver transplantation and post-operative care of these very small patients in our institution. Liver transplantation for infants can be technically challenging.

Biliary Atresia ; surgery ; therapy ; Child, Preschool ; Critical Care ; methods ; Humans ; Infant ; Liver Transplantation ; Living Donors ; Parenteral Nutrition ; Postoperative Care ; methods ; Retrospective Studies ; Treatment Outcome