[Role and progress of allogeneic hematopoietic stem cell transplantation in the treatment of refractory/relapsed diffuse large B-cell lymphoma].

Diffuse large B-cell lymphoma (DLBCL) is the most common type of non Hodgkin's lymphoma. The current treatment plan can significantly improve the prognosis of patients, but about 30%-40% of DLBCL patients still experience drug resistance and relapse after treatment. For patients with refractory/relapse DLBCL, clinical treatment remains difficult and their prognosis is poor. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains one of the most important curative methods for refractory/relapse DLBCL patients. This article will review the role and progress of allo-HSCT in the treatment of refractory/relapse DLBCL.

[Relationship between treatment and prognosis in patients with late-onset severe pneumonia after allogeneic hematopoietic stem cell transplantation].

OBJECTIVE: To explore the relationship between drug treatment and outcomes in patients with late-onset severe pneumonia (LOSP) after allogeneic stem cell transplantation (allo-SCT). METHODS: We retrospectively analyzed the effects of the initiation time of treatment drugs, especially antiviral drugs and glucocorticoids on the clinical outcomes in 82 patients between January 2016 and August 2021 who developed LOSP after allo-SCT in Peking University People's Hospital. Univariate analysis was performed by Mann-Whitney U test and χ2 test, and multivariate analysis was performed by Logistic regression. When multiple groups (n>2) were involved in the χ2 test, Bonferroni correction was used for the level of significance test. RESULTS: Of all 82 patients in this study, the median onset time of LOSP was 220 d (93-813 d) after transplantation, and the 60-day survival rate was 58.5% (48/82). The median improvement time of the survival patients was 18 d (7-44 d), while the median death time of the died patients was 22 d (2-53 d). Multivariate analysis showed that the initiation time of antiviral drugs from the onset of LOSP (< 10 d vs. ≥10 d, P=0.012), and the initiation time of glucocorticoids from antiviral drugs (< 10 d vs. ≥10 d, P=0.027) were the factors affecting the final outcome of the patients with LOSP at the end of 60 d. According to the above results, LOSP patients were divided into four subgroups: group A (antiviral drugs < 10 d, glucocorticoids ≥10 d), group B (antiviral drugs < 10 d, glucocorticoids < 10 d), group C (antiviral drugs ≥10 d, glucocorticoids ≥10 d) and group D (antiviral drugs ≥10 d, glucocorticoids < 10 d), the 60-day survival rates were 91.7%, 56.8%, 50.0% and 21.4%, respectively. CONCLUSION: Our study demonstrated that in patients who developed LOSP after allo-SCT, the initiation time of antiviral drugs and glucocorticoids were associated with the prognosis of LOSP, and the survival rate was highest in patients who received antiviral drugs early and glucocorticoids later. It suggested that for patients with LOSP of unknown etiology should be highly suspicious of the possibility of a secondary hyperimmune response to viral infection.

Protective effects ROS up-regulation on premature ovarian failure by suppressing ROS-TERT signal pathway.

OBJECTIVE: Premature ovarian failure (POF) refers to the condition of pre-onset ovarian function failure, and is one commonly occurred disease in gynecology. Its pathogenic mechanism, however, is still unclear. Early study found decreased activity of telomerase reverse transcriptase (TERT). As an important factor to suppress TERT, oxidative stress has not been studied in POF. We, thus, investigated the role of reactive oxygen species (ROS)-TERT in POF. MATERIALS AND METHODS: Rat POF model was induced by a single intraperitoneal injection of cyclophosphamide plus 12 mg/kg busulfan. Level of follicle stimulating hormone (FSH) and inhibin B was measured by enzyme-linked immunosorbent assay (ELISA), along with hematoxylin and eosin (HE) staining to confirm successful generation of models. Western blot was applied to measure TERT expression, and N-acetyl-cysteine (NAC) or TERT small interfere RNA (siRNA) was injected to suppress ROS or TERT level, followed by HE staining to observe POF condition. RESULTS: In POF model, ovary tissues showed atrophy, less follicles, and more follicular atresia, plus mesenchymal hyperplasia. FSH and inhibin B level were significantly up-regulated and down-regulated, respectively (p<0.05). In POF rat, ROS level was elevated (p<0.05) whilst TERT level was decreased. NAC inhibited ROS level and enhanced TERT expression. In contrast, TERT siRNA further aggravated POF condition. CONCLUSIONS: ROS up-regulation inhibits TERT expression, suppresses TERT activity and facilitates POF. The ROS-TERT pathway may work as the target for treating POF.

Overexpression of TAFI promotes epithelial mesenchymal transition in endometriosis.

OBJECTIVE: Endometriosis is a disease that occurs in women. Thrombin-activated fibrinolytic inhibitor (TAFI) is mainly secreted by stem cells and acts as a regulatory role in the body. Epithelial leaf transition plays a leading role in cell growth and invasion. Our study focuses on the mechanism of TAFI in patients with endometriosis. PATIENTS AND METHODS: The expression of TAFI was determined by immunohistochemistry. Reverse transcriptase-polymerase chain reaction (RT-PCR) served to detect the expression of TAFI and the effect of TAFI on overall survival (OS) and progression-free survival (PFS) levels. The changes of primary cytology in patients with endometriosis were observed under a microscope. The cell source was further determined by immunofluorescence labeling of vimentin and cytokeratin, and the expression of TAFI was detected by Western-blot. 3-4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and cell invasion assay were utilized to detect the viability and aggressiveness of cells after epithelial mesenchymal transition (EMT). RESULTS: TAFI was overexpressed in endometriosis tissues and no expression of TAFI was found in normal tissues, which is consistent with RT-PCR results. TAFI overexpressed endometriosis patients had low levels of overall OS and PFS. There were statistically significant differences. Cell morphology shows that endometriosis primary cells are mainly composed of epithelial cells and fibroblasts. Immunofluorescence assay showed that vimentin and cytokeratin were expressed in cells, and the expression of TAFI was detected by Western-blot. Compared with normal tissues, TAFI was considerably higher in patients with endometriosis. The results of Western-blot and RT-PCR showed that the expression of TAFI was significantly increased in patients with endometriosis and the cell proliferation and cell invasion were significantly accelerated. CONCLUSIONS: Our results show that TAFI is highly expressed in endometriosis and causes EMT, which accelerated the cell proliferation and cell invasion. Snail is an inhibitor of E-cadherin, which may participate in metastasis and invasion of endometriosis by mediating EMT. So, we suspect that Snail controls the occurrence of the EMT and then affects the cell metastasis and invasion, which requires further verification.

[To evaluate the clinicopathologic characteristics and outcome of tumor cells spreading through air spaces in patients with adenocarcinoma of lung].

Objective: To investigate the clinicopathologic features, molecular characteristics and prognosis of spread through air space (STAS) in patients with adenocarcinoma of the lung. Methods: Two hundred and eighty-eight lung adenocarcinoma patients with complete clinicopathologic and follow-up data were included. The patients were divided into STAS positive (178 cases) and negative (110 cases) groups.EGFR and KRAS gene mutations were detected by amplification refractory mutation system (ARMS), and ALK and ROS1 gene fusion were detected by fluorescence in situ hybridization method. The relationship between STAS and clinicopathologic, molecular features, and patient outcome was analyzed. Results: STAS was present in 61.8%(178/288) of lung adenocarcinomas. The positive rate of STAS in tumors >3 cm was significantly higher than that in tumors ≤3 cm (P=0.009), and was significantly higher in tumors with pleural invasion (P<0.01), venous invasion (P<0.01), lymphatic invasion (P<0.01), perineural invasion (P=0.029) and tumors with necrosis (P<0.01). STAS was also correlated with tumor recurrence (P<0.01) and advanced pathologic TNM stage (P=0.002). There was no significant correlation with patients' gender, age and smoking history. Histologically, STAS was present in 58.0%(91/157), 67.6%(50/74), 2/6, 64.3%(27/42) and 8/9 of acinar, papillary, lepidic, solid and micropapillary adenocarcinomas, respectively. In addition, the positive rates of STAS in tumor with micropapillary (>5%) and without micropapillary pattern were 80.9%(55/68) and 55.9%(123/220), respectively (P<0.01). STAS was significantly higher in EGFR negative group (P=0.034), ALK gene rearrangement group (P=0.003) and ROS1 gene rearrangement group (P=0.012), but there was no significant correlation with KRAS mutation. Univariate survival analysis showed that patients with STAS had a shorter progression-free survival (PFS, P<0.01) and overall survival (P=0.013). Multivariate analysis confirmed that STAS was an independent predictor of PFS in lung adenocarcinoma patients (HR: 2.749, 95%CI: 1.550-4.876, P=0.001). Conclusions: The presence of STAS in lung adenocarcinoma suggests high risk of recurrence and invasion and is thus an important prognostic factor. In addition, STAS is associated with EGFR mutation, ALK and ROS1 gene rearrangement.

Blood folic acid, vitamin B12, and homocysteine levels in pregnant women with fetal growth restriction.

Deficiencies in nutrients such as folic acid and vitamin B12 may play a role in fetal growth restriction (FGR). However, whether folic acid, vitamin B12, or homocysteine is associated with FGR in Chinese populations remains unclear. This study investigated the relationship between these nutrient deficiencies and FGR in pregnant Chinese women. We selected 116 mother and infant pairs, and categorized the neonates into the FGR, appropriate for gestational age, and large for gestational age groups. Birth weight, body length, head circumference, body mass index (BMI), and Rohrer's body index of the newborns were measured. Serum folic acid, vitamin B12, and homocysteine levels were measured in mothers during the first three days of their hospital stay. Results showed that the FGR group exhibited reduced folic acid and vitamin B12 levels and elevated homocysteine levels than those in the other two groups. Folic acid and vitamin B12 levels were positively correlated with birth weight, head circumference, and BMI, whereas homocysteine level was negatively correlated with these variables. The FGR ratio in the folic acid and vitamin B12 deficiency group was higher than that in the sufficiency group (χ2 = 4.717 and 4.437, P = 0.029 and 0.035, respectively). In addition, elevated homocysteine was associated with FGR (χ2 = 5.366, P = 0.021). In conclusion, we found that folic acid and vitamin B12 deficiency was associated with elevated homocysteine levels, which may increase susceptibility to FGR.

Oxaliplatin-rapamycin combination was superior to mono-drug in treatment of hepatocellular carcinoma both in vitro and in vivo.

The presented study aimed to investigate the antitumor efficacy of combination of oxaliplatin with rapamycin, an mTOR inhibitor, in hepatocellular carcinoma (HCC). The activation status of mTOR pathway was first examined in HCC cell lines HepG2, BEL7402, and HuH7 using Western blotting. Effects of rapamycin, oxaliplatin, and their combination on the proliferation of HCC cells were determined in vitro using MTT assay and in vivo using a nude mice model bearing HepG2 xenografts. Drug-induced cell apoptosis was examined by flow cytometry. Expression of apoptosis-related protein was determined by Western blotting. We observed that mTOR pathway was activated in all three cell lines used in the current study. MTT assay demonstrated that oxaliplatin in combination with rapamycin synergistically inhibited the proliferation of HCC cells. The combination regimen reduced terminal tumor burden more efficiently than the corresponding monotherapy. The percentages of apoptotic cells and the expression levels of apoptosis-related proteins including cleaved caspase-9, -3, and PARP were significantly higher in combination-treatment groups than those in mono-drug-treatment groups. The ratios of Bax/Bcl-2 in cells exposed to both oxaliplatin and rapamycin were significantly increased compared to those in cells subjected to oxaliplatin or rapamycin alone treatment. Results obtained in the presented study suggested that combination of oxaliplatin and rapamycin was superior to mono-drug and may have a potential value in treatment of HCC.

Two new alleles, HLA-B*35:226 and HLA-B*40:233 identified by sequence-based typing in Chinese individuals.

HLA-B*35:226 differs from HLA-B*35:28, and HLAB*40:233 differs from HLA-B*40:01:01, both by a nonsynonymous mutation.

[Expressions and their clinical significance of bcl-2 and nm23 in epithelial ovarian carcinoma].

The expressions of bcl-2 and nm23 protein in epithelial ovarian carcinoma were analyzed by immunohistochemical method. The results were that 1. the positive expression of bcl-2 protein was 60% (27/45) in epithelial carcinoma; the positive expression of bcl-2 protein in Grade I was 31.25% (5/16), in Grade II 57.14% (8/14), in Grade III 93.33% (14/15) and there were significant differences among them (P < 0.05); 2. the positive expression of nm23 protein was 40% (18/45) in epithelial carcinoma; the positive expression of bcl-2 protein in Stage I/II was 57.1% (12/21), in Stage III/IV 25% (6/24), and there was significant difference between them (P < 0.05). We conclude that the expressions of bcl-2 and nm23 protein are associated with tumor differentiation and clinical stage respectively. Analyzing the expressions of bcl-2 and nm23 are beneficial to the prognosis of the ovarian tumor patients.

Clenbuterol treatment increases muscle mass and protein content of tumor-bearing rats maintained on total parenteral nutrition.

Treatment of tumor-bearing (TB) and control rats with the anabolic beta-2 agonist drug clenbuterol (CLE) for 14 days reduced food intake for 4 days initially. Feeding was increased in anorectic TB rats, however, during the last 7 days of drug administration. Since minimal muscle savings were observed in chow-fed TB rats treated with CLE, the anabolic effects of this drug were investigated in a second experiment on TB rats maintained on total parenteral nutrition (TPN). Sixteen days after the subcutaneous transplantation of methylcholanthrene-induced sarcomas rats was begun on a 2-week schedule of TPN. One group of these rats was treated daily for 14 days with CLE, while the remaining rats received injections of saline. Additional groups of TB and nonTB rats were maintained on rat chow for this period and treated with saline. Although TB rats maintained on rat chow or TPN and treated with saline exhibited significantly decreased gastrocnemius muscle weight and protein content, treatment of TB-TPN rats with clenbuterol normalized muscle mass and increased muscle protein content significantly and increased plasma concentrations of branched-chain amino acids. These results indicate that although nutritional support of TB organisms does not result in protein repletion, the addition of an anabolic drug renders the nutritional support highly efficacious.

Tumor-induced alterations in brain neurotransmitter and plasma ammonia concentrations are normalized twenty-four hours after tumor resection.

Tumor-induced anorexia was accompanied by significant elevations in plasma ammonia and lactate and by alteration of the plasma amino acid profile. The brains of anorectic tumor-bearing rats had increased levels of glutamine and most large neutral amino acids. Dopamine and serotonin metabolism were also increased in several brain regions of these rats. Resection of the tumor resulted in the normalization of most of these aberrations in blood and brain within 24 hrs. These results demonstrate a rapid reversal of tumor-induced biochemical alterations shortly after tumor removal and suggest that these aberrations may be secondary to hyperammonemia.

Response of tumor and host to hyperalimentation and antiglutamine treatments.

In two experiments, treatment of tumor-bearing (TB) rats with the glutamine antimetabolite, acivicin, reduced tumor growth during 14 days of total parenteral nutrition (TPN) that supplied 120% and 130% of the nutritional intake of non-TB control rats. The acivicin treatment or combination of acivicin with TPN produced increased carcass weights and decreased tumor/carcass ratios. Significant muscle saving was observed in TB rats receiving TPN or TPN and acivicin as compared to acivicin-treated TB rats maintained on rat chow. Tumor growth was not stimulated by TPN at 130% of ad libitum. intake with a calorie/nitrogen ratio of 102:1. However, when the calorie/nitrogen ratio was increased to 143:1, tumor growth was increased by TPN at 120% of ad libitum. intake. These results suggest that acivicin may prove useful in the stabilization of tumors in situations where tumor growth may be stimulated, such as during TPN.

Cardiovascular and renal actions of endothelin: effects of calcium-channel blockers.

We have evaluated the effects of two calcium-channel blockers, verapamil (VP) and manganese (Mn), on endothelin (EN)-induced changes in systemic and renal function in pentobarbital sodium-anesthetized female rats and male and female dogs. In the rat studies, saline was infused at 24 microliters/min iv with or without (n = 10) two doses of VP (0.02 mg.kg-1.min-1, n = 5; 0.03 mg.kg-1.min-1, n = 3) or Mn (0.5 mg.kg-1.min-1, n = 5) throughout the entire experiment. After surgery, rats were allowed 60 min to stabilize, and three 20-min control clearances were collected. EN (100 ng.kg-1.min-1) was then added to the infusate for 30 min. EN alone caused an increase in mean arterial pressure (MAP) and a decrease in the glomerular filtration rate (GFR). VP at either dose and Mn totally blocked the EN-induced increase in MAP. However, the two calcium-channel blockers had no effect on the renal action of EN; the GFR (in ml/min) decreased (at 30 min of EN infusion) from 2.7 +/- 0.1 (SE) to 0.7 +/- 0.1 (P less than 0.01) in controls, from 2.5 +/- 0.3 to 0.4 +/- 0.3 with the lower dose of VP (P less than 0.01), from 2.3 +/- 0.1 to 0.6 +/- 0.2 with the higher dose of VP (P less than 0.01), and from 1.5 +/- 0.5 to 0.9 +/- 0.6 with Mn (P less than 0.05). Infusion of EN alone (10 ng.kg-1.min-1, n = 5) and EN with VP (50 micrograms/min, n = 3) into the renal artery of dogs reduced renal blood flow from 3.5 +/- 0.4 to 0.9 +/- 0.2 and from 3.6 +/- 0.6 to 1.2 +/- 0.6 ml.g-1.min-1, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Cardiorenal actions of endothelin, Part I: Effects of converting enzyme inhibition.

We have evaluated the effects of endothelin-1, with and without captopril administration, on the circulating concentration of aldosterone in pentobarbital (60 mg/kg) anesthetized rats. Following surgery, rats (N = 5/group) were infused with saline intravenously, at a rate of 0.024 ml/min, with or without captopril (5 mg kg-1 hr-1) administration throughout the entire experiment. All rats were allowed 60 min to stabilize and 3 X 20 min control clearances collected. Endothelin-1 (100 eta kg-1 min-1) was then added to the infusate for 30 min. Plasma aldosterone concentration increased from 60 +/- 7 eta/dl to 171 +/- 14 eta/dl (p less than 0.01) with endothelin alone and from 101 +/- 6 eta/dl to 210 +/- 54 eta/dl (p less than 0.01) in rats treated with endothelin plus captopril. The endothelin-induced increment in blood pressure was not altered by captopril treatment. However, the endothelin-induced decrement in renal function was markedly attenuated in rats treated with captopril. These data demonstrate that endothelin stimulates the release of aldosterone from the rat adrenal and that the angiotensin II is not involved in this response. These data also demonstrate that the endothelin-induced systemic vasoconstriction is not affected by captopril whereas the endothelin-induced changes in renal function are abolished by captopril.