[D-dimer contributes to the diagnosis and prognosis in hepatitis B-related acute-on-chronic liver failure].

Objective: To investigate the diagnostic and prognostic value of D-dimer level in patients with hepatitis B-related acute-on-chronic liver failure (HBV-ACLF). Methods: A total of 142 cases diagnosed with ACLF were randomly selected as research objects in the open cohort using the Chinese Group on the Study of Severe Hepatitis B-ACLF (COSSH-ACLF). Plasma D-dimer levels were compared between patients with ACLF and non-ACLF and patients with different ACLF grades. Survival and death group D-dimer levels were compared with the end points of 28 days and 90 days, respectively. The correlation between D-dimer and other laboratory indicators and prognostic scores were investigated. Area under receiver operating characteristic curve (AUROC) was used to evaluate the D-dimer value for predicting the prognosis of ACLF patients. 125 external ACLF cases were used for validation. A Student t test or Mann-Whitney U test was used to compare continuous measurement data between two groups. Kruskal-Wallis test was used to compare continuous measurement data between multiple groups. Results: Plasma D-dimer levels in the ACLF [2 588.5 (1 142.8, 5 472.8) µg/L] ] and non-ACLF group [1 385.5 (612.0, 3 840.3) µg/L] had a significant difference (P<0.001). ACLF-3 patients had significantly higher D-dimer levels than ACLF-1/2 patients (ACLF-3 vs. ACLF-1, P<0.001; ACLF-3 vs. ACLF-2, P<0.05). Patients who died at 28/90 days had significantly higher D-dimer levels than those whom survived (P<0.001). There was a significant positive correlation between D-dimer level with prothrombin time (PT), international normalized ratio (INR), high-density lipoprotein C, as well as various prognostic scores (COSSH-ACLFs, CLIF-C ACLFs, CLIF-OFs, MELDs). AUROC of D-dimer in predicting the prognosis of ACLF patients at 28 days and 90 days was 0.751 (95% CI: 0.649-0.852) and 0.787 (95% CI: 0.695-0.878), respectively, which did not differ significantly compared with the predictive ability of other scores (P<0.05), and similar results were confirmed by an external validation group of 125 cases. Conclusion: D-dimer level is significantly higher in patients with ACLF, so it is an independent predictor of prognosis at 28 and 90 days.

[Expression and diagnostic value of macrophage inflammatory protein-3α in patients with hepatitis B-related acute-on-chronic liver failure].

Objective: To screen serum protein markers and evaluate their diagnostic application value in hepatitis B-related acute-on-chronic liver failure (HBV-ACLF). Methods: Serum samples of patients with HBV-ACLF, chronic hepatitis B (CHB) and normal healthy volunteers (n = 5/group) were determined by cytokine antibody chip in line with the Chinese Diagnostic Standards Study for HBV-ACLF (COSSH-ACLF) cohort. The differentially expressed proteins significance were identified by microarray analysis and prediction. The preliminary serological markers of HBV-ACLF were screened for diagnosis. The potential markers were determined by enzyme-linked immunosorbent assay (ELISA), area under the receiver operating characteristic curve (AUROC) analysis and liver tissue immunohistochemistry for the diagnosis of HBV-ACLF. Student t-test or Mann-Whitney U test were used to compare the continuous measurement data between the two groups, and analysis of variance and Kruskal-Wallis test were used to compare the continuous measurement data between multiple groups. Results: Cytokine antibody chip preliminary screening results showed that the expression levels of these six cytokines, namely, macrophage inflammatory protein 3α (MIP-3α), hepatocyte growth factor, E-selectin, osteopontin, growth differentiation factor 15 and carcinoembryonic antigen-related cellular adhesion molecule 1 were significantly increased in the HBV-ACLF group. Among them, the expression level of MIP-3α was significantly higher in the HBV-ACLF group (99.6 times higher than CHB group and 146.9 times higher than healthy volunteers' group, respectively, P < 0.0001) as validated by serum ELISA in 132 HBV-ACLF cases, 91 CHB cases and 72 healthy volunteers. AUROC analysis showed that the high expression of MIP-3α could be used as a marker to distinguish patients with HBV-ACLF from CHB. The AUROC was 0.995 (95% CI: 0.990 ~ 1.000), with sensitivity and specificity of 95.5% and. 98.9%, respectively. Immunohistochemistry showed that MIP-3α was positively expressed in HBV-ACLF-derived liver tissues, and negatively expressed in CHB-derived liver and normal liver tissues. Conclusion: Serum MIP-3α level is closely related to the pathological characteristics of HBV-ACLF. Therefore, it may be used as a potential serological marker for the diagnosis of HBV-ACLF.

[Establishment of clinical features and prognostic scoring model in early-stage hepatitis B-related acute-on-chronic liver failure].

Objective: To explore the clinical characteristics and establish a corresponding prognostic scoring model in patients with early-stage clinical features of hepatitis B-induced acute-on-chronic liver failure (HBV-ACLF). Methods: Clinical characteristics of 725 cases with hepatitis B-related acute-on-chronic hepatic dysfunction (HBV-ACHD) were retrospectively analyzed using Chinese group on the study of severe hepatitis B (COSSH). The independent risk factors associated with 90-day prognosis to establish a prognostic scoring model was analyzed by multivariate Cox regression, and was validated by 500 internal and 390 external HBV-ACHD patients. Results: Among 725 cases with HBV-ACHD, 76.8% were male, 96.8% had cirrhosis base,66.5% had complications of ascites, 4.1% had coagulation failure in respect to organ failure, and 9.2% had 90-day mortality rate. Multivariate Cox regression analysis showed that TBil, WBC and ALP were the best predictors of 90-day mortality rate in HBV-ACHD patients. The established scoring model was COSS-HACHADs = 0.75 × ln(WBC) + 0.57 × ln(TBil)-0.94 × ln(ALP) +10. The area under the receiver operating characteristic curve (AUROC) of subjects was significantly higher than MELD, MELD-Na, CTP and CLIF-C ADs(P < 0.05). An analysis of 500 and 390 cases of internal random selection group and external group had similar verified results. Conclusion: HBV-ACHD patients are a group of people with decompensated cirrhosis combined with small number of organ failure, and the 90-day mortality rate is 9.2%. COSSH-ACHDs have a higher predictive effect on HBV-ACHD patients' 90-day prognosis, and thus provide evidence-based medicine for early clinical diagnosis and treatment.

[Clinical and genetic characteristics of young patients with myeloproliferative neoplasms].

Objectives: To investigate the clinical and genetic features of young Chinese patients with myeloproliferative neoplasms (MPN). Methods: In this cross-sectional study, anonymous questionnaires were distributed to patients with MPN patients nationwide. The respondents were divided into 3 groups based on their age at diagnosis: young (≤40 years) , middle-aged (41-60 years) , and elderly (>60 years) . We compared the clinical and genetic characteristics of three groups of MPN patients. Results: 1727 assessable questionnaires were collected. There were 453 (26.2%) young respondents with MPNs, including 274 with essential thrombocythemia (ET) , 80 with polycythemia vera (PV) , and 99 with myelofibrosis. Among the young group, 178 (39.3%) were male, and the median age was 31 (18-40) years. In comparison to middle-aged and elderly respondents, young respondents with MPN were more likely to present with a higher proportion of unmarried status (all P<0.001) , a higher education level (all P<0.001) , less comorbidity (ies) , fewer medications (all P<0.001) , and low-risk stratification (all P<0.001) . Younger respondents experienced headache (ET, P<0.001; PV, P=0.007; MF, P=0.001) at diagnosis, had splenomegaly at diagnosis (PV, P<0.001) , and survey (ET, P=0.052; PV, P=0.063) . Younger respondents had fewer thrombotic events at diagnosis (ET, P<0.001; PV, P=0.011) and during the survey (ET, P<0.001; PV, P=0.003) . JAK2 mutations were found in fewer young people (ET, P<0.001; PV, P<0.001; MF, P=0.013) ; however, CALR mutations were found in more young people (ET, P<0.001; MF, P=0.015) . Furthermore, mutations in non-driver genes (ET, P=0.042; PV, P=0.043; MF, P=0.004) and high-molecular risk mutations (ET, P=0.024; PV, P=0.023; MF, P=0.001) were found in fewer young respondents. Conclusion: Compared with middle-aged and elderly patients, young patients with MPN had unique clinical and genetic characteristics.

[Health-related quality of life and its associated variables in Chinese patients with Philadelphia-negative myeloproliferative neoplasms].

Objectives: To explore health-related quality of life (HRQoL) and identify its associated variables in Chinese patients with Philadelphia-negative myeloproliferative neoplasms (MPNs) . Methods: In this cross-sectional study, anonymous questionnaires were distributed to adult patients with MPNs to assess symptom burden measured by MPN-10 and HRQoL measured by Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30) . Results: The data from 1405 respondents with MPNs, including 645 (45.9%) with essential thrombocythemia (ET) , 297 (21.1%) with polycythemia vera (PV) , and 463 (33.0%) with myelofibrosis (MF) , were analyzed. 646 (46.0%) respondents were male. The median age was 56 (range, 18-99) years. The mean MPN-10 scores were 13.0±12.7, 15.0±14.7, and 21.0±16.6 (P<0.001) , and the physical component summary (PCS) and mental component summary (MCS) scores were 48.0±8.5, 47.0±9.0, and 42.0±10.0 (P<0.001) and 51.0±11.0, 50.0±10.8, and 49.0±11.1 (P=0.002) for respondents with ET, PV, and MF, respectively. Respondents with MF reported the lowest score of physical functioning, role functioning, emotional functioning, cognitive functioning, social function, and global health status (all P<0.01) and the highest score of fatigue, pain, dyspnea, appetite loss, diarrhea, and financial problems (all P<0.05) in EORTC QLQ-C30. Multivariate analyses revealed that higher MPN-10 scores were significantly associated with lower PCS (-0.220 to -0.277, P<0.001) and MCS (-0.244 to -0.329, P<0.001) scores; increasing age (-1.923 to -4.869; all P<0.05) , lower PCS score. Additionally, comorbidity (ies) , symptom at diagnosis, splenomegaly, anemia, unknown driver gene, and higher annual out-of-pocket cost were significantly associated with lower PCS and/or MCS scores. However, age ≥ 60 years, urban household registration, concomitant medication, and receiving ruxolitinib therapy in respondents with MF were associated with higher MCS scores. Weak correlations were found between MPN-10 score (except the subscale of appetite loss and constipation) and EORTC QLQ-C30 score in majority of subscales in respondents with ET (|r| = 0.193-0.457, all P<0.001) , PV (|r| = 0.192-0.529, all P<0.01) , and MF (|r| = 0.180-0.488, all P<0.001) , respectively. Conclusions: HRQoL in patients with MPN was significantly reduced, especially in patients with MF. Sociodemographic and clinical variables were significantly associated with the HRQoL in patients with MPNs.

[Variables associated with BCR-ABL kinase domain mutation in TKI-resistant patients with chronic myeloid leukemia].

Objectives: To explore BCR-ABL kinase domain mutation profiles and clinical variables associated with them in tyrosine kinase inhibitor (TKI) -resistant patients with chronic myeloid leukemia (CML) . Methods: Imatinib-, nilotinib-, and/or dasatinib-resistant patients with CML who screened BCR-ABL mutation (s) in Peking University People's Hospital between June 2001 and September 2019 were retrospectively reviewed. BCR-ABL mutation was analyzed by Sanger sequencing. Binary logistic regression model was built to identify independent clinical variables associated with developing BCR-ABL mutation (s) . Results: Data of 1 093 TKI-resistant cases in 804 patients who experienced resistance to imatinib (n=576, 52.7%) , nilotinib (n=238, 21.8%) , and dasatinib (n=279, 25.5%) were analyzed. In total, 291 (50.5%) imatinib-, 152 (63.9%) nilotinib-, and 160 (57.3%) dasatinib-resistant cases developed BCR-ABL mutation (s) . T315I mutation was the most frequent mutation detected in imatinib-, nilotinib-, and dasatinib-resistant cases, accounting for 12.3%, 27.3%, and 34.1%, respectively. Y253F/H (7.5%) and F359V/C/I (5.6%) were the mutation detected in ≥5% imatinib-resistant cases; F359V/C/I (12.2%) , Y253F/H (11.8%) , and E255K/V (10.5%) in nilotinib-resistant cases; and F317L/V/I/C (11.5%) and E255K/V (5.4%) in dasatinib-resistant cases. In multivariate analyses, no TKI dose reduction or discontinuation of TKI therapy was the common variable associated with developing BCR-ABL mutation (s) . Other variables associated with developing BCR-ABL mutation (s) in imatinib-, nilotinib-, or dasatinib-resistant cases included male gender, younger age, no comorbidity, advanced phase before starting current TKI therapy, longer interval from diagnosis to starting current TKI therapy, acquired resistance, and TKI resistance due to progression to advanced phase or hematologic failure. In addition, interval from TKI failure to BCR-ABL mutation detection, starting initial TKI therapy to TKI failure, and starting current TKI therapy to TKI failure were associated with the frequency of developing BCR-ABL mutation. Dasatinib and nilotinib use and acquired resistance were identified to be associated with the development of T315I mutation in multivariate analyses. Conclusions: More than half of TKI-resistant CML patients developed BCR-ABL mutation (s) by Sanger sequencing. T315I mutation was the most frequently detected. Clinical variables significantly associated with developing BCR-ABL mutation (s) should be used not only as basis for the choice of subsequent TKIs but also the understanding of TKI-resistant mechanisms.

[Myelopathy and cervical spino-stenosis].

From 1986 through 1990, among the cases of traumatic myelopathy of the cervical spine admitted in our hospital, 31 cases complicated with sagittally narrowed vertebral canal were reviewed. The stenotic cervical canal would be susceptible to spinal cord damage, such as a very minute trauma or violent hyperextension motion of the neck in the aged patients would lead to the cord damage. This type of cord injury was characterized by very little symptoms and signs in the neck, normal or only minimal bony damage seen on roentgenograms or CT scans, even though the spinal cord damage was very severe. With non-operative treatment the recovery chance was frequently very poor or rather, it would be getting worse gradually. Surgical treatment such as expansion canal laminoplasty could give a better result.

[Early diagnosis of atlanto-axial tuberculosis].

Spinal tuberculosis at the atlanto-axial level is often overlooked or misdiagnosed for a considerable period of time, until when there has developed subluxation or other obvious complications. In order to make early diagnosis of this lesion, we analysed 30 such patients of our own and found that the positive X-ray film findings, such as thickening and widening of the soft tissue shadow of the retropharyngeal wall could hardly be concerned as an early sign, subluxation of the joint and destruction of bone were definitely quite a late occurrence. While long lasting pain in the upper cervical region and limitation of motions of the neck were constantly present prior to any other positive findings. Complaints of these subjective symptoms, particularly in young person, we would emphasize, are the early signals of tuberculosis and should be cared until being proved otherwise.

Effects of glutamine on tumor growth and apoptosis of hepatoma cells.

AIM: To explore the effects of glutamine on growth and apoptosis of hepatoma cells. METHODS: Mice inoculated with hepatoma cell (H22) suspension subcutaneously at right axilla were orally administered with glutamine (GLN) solution. Human hepatoma cell culture (SMMC-7721) was treated with different concentrations of GLN solution. The content of malondialdehyde (MDA) and nitric oxide (NO) was detected in mice plasma and cell culture, and that of glutathione (GSH) was decected in cells. The inoculated tumor's growth in the mice and hepatoma cells' proliferation and apoptosis were observed. RESULTS: When mice were administered orally with GLN solution (300 mg/kg), the growth of inoculated hepatoma was suppressed in the mice. When different concentrations of GLN solution were added in human hepatoma cell culture, the hepatoma cells' proliferation was inhibited and cells were induced to apoptosis, which was dependent on GLN concentration; meanwhile the contents of NO rose both in mice plasma and in cell culture, however MDA contents were slightly lowered in both, and the activity of GSH increased in the cells which had been ultrasonically shattered. CONCLUSION: Hepatoma cell apoptosis and tumor growth inhibition by GLN may be associated with its antioxidative activity and its intervention in hepatoma cell proliferation, and simultaneous release of NO.

Biliary tract reconstruction using an autologous vein graft in rats.

Numerous biologic and synthetic materials have been used with limited success as an interposed graft to repair segmental common bile duct (CBD) defects. The authors report here that an autologous vein graft can be successfully used to correct a CBD deficit contingent on accurate microsurgical technique immediate stenting and rapid graft vascularization. Thirty Sprague-Dawley rats underwent laparotomy and the experimental group (n=25) had a 3-mm segment of the CBD excised. The CBD defect was repaired using an interposed femoral vein graft aided by a plastic stent. The control group (n=5) had the CBD cut and repaired by means of primary anastomosis. The experimental group was subdivided into three sub-groups each examined at three different postoperative intervals: 1, 4 and 12 weeks. The results showed that inflammation was apparent in the venous wall following the first postoperative week. A progressive loss of the vascular endothelium and replacement with the columnar epithelium typical of the CBD was seen in the vein graft. Nineteen of the 25 experimental rats (76 percent) of the animals survived without complication from the surgery and there were no abnormalities in the liver biochemical tests of these animals. Any biliary tract obstruction that developed was attributed to dislocation of the stent leading to collapse of the vein graft (experimental group), or constriction of the anastomosis (control group). This study demonstrates that biliary tract reconstruction using an autologous vein graft can be successfully performed in a rat model of CBD repair. The application of this method to the clinical setting is also discussed.

Saphenous neurocutaneous island flap model in the rat: evaluation of vascular supply.

Neurocutaneous flaps are utilized routinely in reconstructive surgery and even more so during the past decade. In this study, the vascular supply of the neurocutaneous flap in the rat model is presented and evaluated. Thirty-six flaps (3.5x3 cm2) were dissected on the medial aspect of the leg based on a pedicle of the saphenous nerve, saphenous artery, great saphenous vein, and the surrounding fascial tissues. Animals in the experiment were divided into five groups with different circulatory patterns of pedicle dissections. In group I (N = 12), the pedicle artery, vein, nerve, and fascia were preserved. In group II (neurocutaneous flap model, N = 24), the saphenous artery was transected and the vein, nerve, and fascia were preserved. In group III (intraneural vascular plexus circulation pattern, N = 12), the saphenous artery and the fascia were transected. In group IV (extraneural vascular plexus circulation pattern, N = 12), the saphenous artery and nerve were transected. In group V (N = 12), the entire pedicle was transected completely. Flap survival was evaluated grossly on postoperative day 7. All flaps survived in group I, but in group II 19 of 24 flaps survived completely, 3 of 24 had partial necrosis, and 2 of 24 were completely necrotic. Complete necrosis was observed in all group III flaps. In group IV, 6 of 12 flaps survived completely, 3 of 12 flaps survived partially, and 3 of 12 flaps were necrotic (p<0.05 vs. group I). Only one flap with partial necrosis survived in group V. In group II, the average survival area was not significantly different from group I (p>0.05). In conclusion, the saphenous neurocutaneous flap in the rat is a reliable microsurgical model. The saphenous neurocutaneous flap is commonly supplied by both the intraneural and extraneural vascular plexus, and although the latter is more important, neither provides sufficient vascular supply on its own.

Effect of hyperbaric oxygen on microvascular anastomosis healing and patency in the rat.

Recent evidence has shown that changes in blood viscosity and marked increases in both platelet count and fibrinogen concentration occur after exposure to hyperbaric oxygen (HBO). The purpose of the present study was to address whether repeated HBO therapy would increase either hematocrit or platelet count, and to determine if these changes would influence the patency of the microvascular anastomosis, as well as anastomotic healing. Eighty femoral arteries from 40 rats were divided into four groups (n = 10), and arterial anastomoses were performed on normal arteries in the first two groups, and on crushed arteries in the second two groups. One of the normal artery groups and one of the crushed artery groups received repeated HBO treatments for 5 days. Anastomotic patency rates, platelet count, hematocrit, and breaking strength were recorded. Among the results of this study were that: (1) there was no difference in patency rate following HBO treatment in both the normal and crushed artery groups; (2) platelet count was significantly higher following HBO treatment in the groups with the undamaged arteries; (3) breaking strength was significantly increased following HBO treatment in the groups with the crushed arteries. The authors concluded that HBO therapy may be useful in improving the healing of microvascular anastomoses in vessels that have undergone crush injury. Although HBO treatment results in an increased platelet count, this does not influence anastomotic patency.

Quadriceps femoris muscle flap: largest muscle flap model in the rat.

The purpose of this paper is to present a new muscle flap in the rat: the quadriceps femoris muscle flap based on a pedicle consisting of the femoral vessels. In order to establish the anatomic details of this model, seven rats were explored bilaterally, and the regional anatomy of the thigh was examined. The technical aspect of the model was established by the unilateral harvesting of 18 quadriceps femoris muscles. Findings were that this muscle is primarily supplied by a muscular branch originating at the superficial circumflex iliac artery. The average muscle weight was 6 g and the average pedicle length with femoral vessels was 6 mm. Eight of the harvested flaps were transplanted to the contralateral thigh, and the pedicle was anastomosed to the femoral vessels. The other ten flaps were resutured back to their beds. At 72 hr postoperatively, all flaps were viable with the exception of one of the transplanted flaps which was found to be necrotic. The quadriceps femoris muscle flap is technically both a reliable and simple model. With an average weight of 6 g, this flap is by far the largest described in the rat, and offers a convenient model for testing flap-related techniques and outcomes.

Development of a mouse limb transplantation model.

A tremendous amount of research has been dedicated to laying the groundwork that will eventually lead to successful limb transplantation in humans. Limb transplantation in animal models has also been widely used for evaluating composite tissue allografts and various immunosuppressive regimens. Currently, there is no mouse model of limb transplantation. Such a model is attractive because it would allow investigators to apply the well-defined genetic characteristics of the mouse to the challenging field of limb transplantation. In this study, 12 mice underwent orthotopic hind limb transplantations using end-to-end anastomoses of the femoral vessels. The success rate of this surgical procedure was 83%, with 10 of the 12 limbs surviving. Experimental devices, operative procedures, and the major elements of success are discussed.

Effects of VEGF administration following ischemia on survival of the gracilis muscle flap in the rat.

The incidence of free flap transplantation failure is only 3% to 5%, yet still occurs in cases in which the flap suffers prolonged ischemia. The purpose of the current study was to determine the effects of vascular endothelial growth factor (VEGF)--a potent angiogenic agent with a suspected role in the protection of endothelium--on flap survival in a model of ischemia-reperfusion injury. The model chosen was the rat gracilis muscle flap. A total of 36 adult male Sprague-Dawley rats were divided into three groups (N = 12). One experimental group received VEGF treatment and the other received heparin. A third group was treated with saline and served as the control. The gracilis muscle flap was dissected and isolated based on a vascular pedicle originating at the femoral vessels. Following 3.75 hours of ischemia, induced by clamping the femoral vessels, either VEGF, heparin, or saline was infused directly into the pedicle of the flap via a cannula. The flaps were evaluated both grossly and histologically after 72 hours of reperfusion. Eleven of the 12 flaps from the VEGF group survived, whereas the survival rate was 6 of 12 and 5 of 12 flaps for the heparin- and saline-treated groups respectively. Flap survival was significantly greater in the VEGF-treated group compared with the heparin- and saline-treated groups (p < 0.025, p < 0.01 respectively). Furthermore, there was no significant difference between the heparin and saline groups. These results indicate that VEGF plays a role in reducing the damage that occurs in ischemia-reperfusion injury, and that the use of VEGF holds promise as a potential therapy for increasing flap survival.