[Clinical value of focused ultrasound ablation surgery in the treatment of abdominal wall endometriosis].

Objective: To explore the effectiveness and safety of focused ultrasound ablation surgery (FUAS) for abdominal wall endometriosis. Methods: From November 2019 to October 2022, a total of 34 patients with abdominal wall endometriosis who underwent FUAS were collected, and their clinical features, imaging features, intraoperative treatment and side effects after treatment were analyzed retrospectively, and the improvement of symptoms and re-intervention were followed up. Results: (1) Characteristics of clinical data: the average age of 34 patients with abdominal wall endometriosis was (32.8±3.8) years old. The largest diameter of the lesion was 48 mm, and the median lesion diameter was 24 mm. Thirty cases (88%, 30/34) had moderate to severe periodic pain in abdominal incision before FUAS. All patients were diagnosed by preoperative magnetic resonance imaging, including 19 cases (56%, 19/34) of superficial type, 8 cases (24%, 8/34) of intermediate type and 7 cases (21%, 7/34) of deep type. (2) FUAS treatment parameters: ablation was completed with average operation time of (64±18) minutes, average sonication time was (385±108) s, (103±11) W of average power, (38 819±16 309) J of average total energy, the average treatment area volume of (3.11±1.42) cm3, and (377.79±106.34) s/h of average treatment intensity. (3) Efficiency: the pain of patients after FUAS was significantly relieved, and the pain scores of patients after 1 month, 3 months, 6 months and 1 year after FUAS were significantly decreased (Z=-4.66, -5.13, -5.11 and -4.91, all P<0.01). One year after FUAS, the near relief and effective pain relief rate was 74% (25/34), and the clinical effective rate was 85% (29/34). Five patients recurred after one year, including 3 patients who underwent abdominal wall endometriosis lesion resection and 2 patients who received drug treatment. One month after FUAS, the size of the lesion did not change significantly compared with that before FUAS (P>0.05), and the size of the lesion decreased significantly after FUAS at 3 months, 6 months and 1 year (Z=-2.15, -2.67 and -3.41, all P<0.05). It has no difference in pain relief among different types (P>0.05), but has significant difference in focus reduction among three types (P<0.01). (4) Safety: there were 34 cases (100%, 34/34) of skin burning sensation, 19 cases (56%, 19/34) of pain in the treatment area and 2 cases (6%, 2/34) of hematuria. All patients got better after corresponding treatments. Conclusion: FUAS is safe and effective for the treatment of abdominal wall endometriosis, which has clinical application value.

[Effects of free superficial peroneal artery perforator flap in repairing small and medium-sized thermal crush injury wounds in the hand].

Objective: To investigate the surgical methods and clinical effects of free superficial peroneal artery perforator flap in repairing small and medium-sized thermal crush injury wounds in the hand. Methods: A retrospective observational study was conducted. From August 2018 to December 2021, 12 patients (19 wounds) with small and medium-sized thermal crush injury in the hand who met the inclusion criteria were hospitalized in Suzhou Ruihua Orthopaedic Hospital, including 5 males and 7 females, aged from 30 to 54 years. The area of the wound was from 2.5 cm×2.0 cm to 14.0 cm×3.5 cm, and all the wounds were repaired by using free superficial peroneal artery perforator flaps from lower leg on one side (including single flap, multiple flaps, and multiple flaps with one pedicle resected from the same donor site). The area of the flap was from 3.5 cm×3.0 cm to 16.0 cm×4.0 cm. The wound in the donor site was sutured directly. The vascular crisis and survival of the flap were observed after operation. The texture, appearance, color, hyperpigmentation, sensation, and two-point discrimination of the flap repaired area were followed up, as well as the hyperplasia of scar and pain condition in the donor and recipient sites. At the last follow-up, the curative effect of flap repair was evaluated by the comprehensive evaluation scale, and the extension and flexion functions of the reserved digital joint were evaluated by the total active movement systematic evaluation method recommended by American Academy for Surgery of Hand. Results: One flap developed arterial crisis on the first day after operation but survived after timely exploration. The other 18 flaps survived successfully after operation. Follow-up of 4 to 24 months after operation showed good texture and appearance in the flap repaired area; the color of the flap repaired area was similar to that of the normal skin around the recipient site, without pigmentation; the protective sensation was restored in all cases, but there was no two-point discrimination; there was no obvious hypertrophic scarring or pain in the donor or recipient site. At the last follow-up, the curative effect of flap repair was evaluated with 3 flaps being excellent and 16 flaps being good; the extension and flexion functions of the reserved digital joint were also assessed, being excellent in 8 fingers, good in 9 fingers, and fair in 2 fingers. Conclusions: The blood supply of superficial peroneal artery perforator flap is sufficient and reliable, and multiple flaps of this type or multiple flaps with one pedicle can be resected from one donor site. The use of this flap to repair small and medium-sized thermal crush injury wounds in the hand results in minimal damage to the donor area, and good postoperative appearance and texture of the flap.

[Clinical effects of proximal ulnar artery perforator flap combined with iliac bone graft in the reconstruction of subtotal thumb or finger defects].

Objective: To explore the clinical effects of proximal ulnar artery perforator flap combined with iliac bone graft in the reconstruction of subtotal thumb or finger defects. Methods: A retrospective observational study was conducted. From August 2016 to August 2019, 7 patients with thumb or finger defects caused by mechanical damage who met the inclusion criteria were admitted to Ruihua Affiliated Hospital of Soochow University, including 6 males and 1 female, aged 46 to 58 years. Their length of fingers was repaired with iliac bone, with length of 2.0 to 3.0 cm. After the bone graft, the skin defect area of the affected finger ranged from 2.8 cm×2.2 cm to 6.0 cm×3.2 cm. Then the free proximal ulnar artery perforator flap with area of 3.0 cm×2.4 cm to 6.5 cm×3.5 cm was used to cover the wounds. The wounds in donor sites of iliac crest and flap were directly sutured. The survival of flap in one week post surgery and the donor site wound healing in 2 weeks post surgery were observed, respectively. During the follow-up, the appearance and sensory function of the affected finger, bone healing, and scar hypertrophy of wound in the donor site were observed and evaluated. At the last follow-up, the functional recovery of the affected finger was evaluated with trial standard for the evaluation of functions of the upper limbs of the Hand Surgery Society of Chinese Medical Association. Results: In one week post surgery, all the flaps survived. In 2 weeks post surgery, the iliac bone and the wounds in forearm donor site healed. During the follow-up of 5 to 13 months, the flap was good in appearance, without obvious pigmentation; the sensory recovery reached level S2 in 5 patients and S0 in 2 patients; all the grafted iliac bones were bony union without obvious resorption; the wounds in donor site healed well, with only mild scar formation. At the last follow-up, the shape of the reconstructed finger was close to the healthy finger, and the functional evaluation results were excellent in 3 cases and good in 4 cases. Conclusions: The use of proximal ulnar artery perforator flap combined with iliac bone graft to reconstruct subtotal thumb or finger can partially restore part of the appearance and function, with less damage to the donor site. It is a good choice for patients who have low expectations of appearance and function for the reconstructed finger.

[Clinical and prognostic features of ovarian endometrioid carcinoma with synchronous endometrial lesions].

Objective: To compare the clinical and prognostic characteristics of ovarian endometrioid carcinoma (OEC) patients with synchronous endometrial lesions and patients with pure OEC. Methods: A retrospective review of the medical records of patients received initial treatment and a postoperative pathological diagnosis of OEC at Peking University People's Hospital between August 1998 and December 2017 were performed. According to the inclusion criteria, a total of 56 patients with OEC were included in the study, including 13 patients concurrent with simultaneous endometrial lesions (Group A) and 43 patients with pure OEC (Group B). Results: Patients with synchronous endometrial lesions accounted for 23% (13/56). Mean age of Group A at diagnosis was (44.9±8.3) years old, 2/13 of patients were postmenopausal, and no one had a history of hypertension, the first symptom of 5/13 people was irregular vaginal bleeding. Mean age of Group B patients at diagnosis was (52.7±10.2) years old, 53% (23/43) of patients were postmenopausal, and 28% (12/43) patients had the history of hypertension, the first symptom of 4 (9%, 4/43) people was irregular vaginal bleeding. The differences of age, menopause status, history of hypertension and initial symptoms between the two groups were statistically significant (all P<0.05). There were no significant differences in fertility history, dysmenorrhea history, age of menarche, history of endometriosis, preoperative and postoperative CA125 level, International Federation of Gynecology and Obstetrics (FIGO) stage, tumor grade, metastatic site and platinum-based chemotherapy drug resistance between the two groups (all P>0.05). The overall 5-year survival rate of OEC patients was 91.6%, and the overall 5-year progression-free survival rate was 76.6%. Among them, the 5-year survival rate of the OEC concurrent with simultaneous endometrial lesions group was 80.2%, and the pure OEC group was 93.4%; the 5-year progression-free survival rate of the OEC concurrent with simultaneous endometrial lesions group was 74.1%, and the 5-year progression-free survival rate of the pure OEC group was 77.3%. There were no significant differences between the two groups (all P>0.05). Multivariate analysis showed that the independent factors for the prognosis of OEC patients were FIGO stage (P=0.006) and residual lesion size (P=0.020). Conclusions: OEC patients have a high proportion of simultaneous endometrial lesions. OEC with simultaneous endometrial lesions are younger than patients with pure OEC. Synchronous endometrial lesions do not affect the prognosis of patients with OEC.

Long non-coding RNA DANCR upregulates IGF2 expression and promotes ovarian cancer progression.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long non-coding RNA DANCR upregulates IGF2 expression and promotes ovarian cancer progression, by Y.-Q. Gao, H.-Y. Cheng, K.-F. Liu, published in Eur Rev Med Pharmacol Sci 2019; 23 (9): 3621-3626-DOI: 10.26355/eurrev_201905_17785-PMID: 31114986" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/17785.

[Preliminary study of PD-1 inhibitor in the treatment of drug-resistant recurrent gestational trophoblastic neoplasia].

Objective: To investigate the therapeutic effect of programmed cell death receptor 1 (PD-1) inhibitor in drug-resistant recurrent gestational trophoblastic neoplasia (GTN). Methods: Clinicopathological features, previous treatments, PD-1 inhibitor treatment and prognosis of 8 patients with drug-resistant recurrent GTN treated with PD-1 inhibitor pembrolizumab， in Peking Union Medical College Hospital of Chinese Academy of Medical Sciences from August 2018 to June 2019 were collected and retrospectively analyzed. Results: (1) Clinicopathological features: the average age of onset of 8 GTN patients was 32.9 years old (31-39 years old); pathological types: choriocarcinoma in 7 cases, epithelioid trophoblastic tumor in 1 case. International Federation of Gynecology and Obstetrics (FIGO) stages: stage Ⅲ in 5 cases, stage Ⅳ in 3 cases; FIGO score: 4 patients with 7-12 points (high-risk group) and 4 patients with ≥13 points (ultra high-risk group). All the 8 patients had lung metastasis, 2 patients with brain metastasis, 1 patient with kidney and 1 patient with intestinal metastasis. (2) Previous treatments: ① Chemotherapy: 8 patients with GTN received an average of 21.1 courses (5-30 courses) of chemotherapy; the main route was systemic intravenous chemotherapy. One patient received intrathecal methotrexate chemotherapy due to brain metastasis. ② Surgery: 7 of 8 patients with GTN received surgical treatment, including 5 cases of pelvic surgury, 6 cases of pulmonary lobectomy and 1 case of right hemicolectomy. ③ Radiotherapy: 2 of 8 patients with GTN received radiotherapy, among which 1 patient received radiotherapy for lung for 8 times due to lung metastasis, and the other one received radiotherapy for lung, right sacroiliac joint and skull for a total of 22 times. (3) PD-1 inhibitor treatment: 8 patients with GTN received PD-1 inhibitor treatment with a mean course of 9 (2-12 courses). Six patients appeared Ⅰ-Ⅱ grade of immune related adverse events (AE), and no severe AE occurred. (4) Prognosis: after 2-3 courses of PD-1 inhibitor treatment, serum ß-hCG level came to normalization in 4 patients. They were followed up for 2-7 months without any recurrence after 0-9 courses of consolidation treatment. One patient received 12 courses of PD-1 inhibitor treatment. The serum ß-hCG level normalized after the 6th courses but increased 1 months later, and then received bevacizumab treatment due to the progression of the disease. The remaining 3 patients received other chemotherapy regiments due to disease progression during PD-1 inhibitor treatment. Conclusions: PD-1 could be used as a remedial treatment for drug-resistant recurrent GTN, with a high effective rate and relatively mild AE. However, more cases need to be accumulated clinically and efficacy should be comprehensively evaluated in combination with pathology and immunohistochemical examination.

Long non-coding RNA DANCR upregulates IGF2 expression and promotes ovarian cancer progression.

OBJECTIVE: Recent studies have revealed the important role of long non-coding RNA (lncRNAs) in the development of malignant tumors. In this work, we explored the exact role of lncRNA DANCR in ovarian cancer progression and the underlying mechanism. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect DANCR expression in both ovarian cancer cells and tissue samples. Subsequently, cell proliferation assay and transwell assay were conducted. Furthermore, the underlying mechanism was explored via qRT-PCR and Western blot assay. RESULTS: The expression of DANCR in ovarian cancer samples was significantly higher than that of the corresponding normal tissues. After DANCR overexpression in vitro, the proliferation, invasion and migration of ovarian cancer cells were markedly promoted. In addition, both the mRNA and protein expression levels of insulin-like growth factor 2 (IGF2) were remarkably upregulated after DANCR overexpression. Furthermore, the results found that the expression level of IGF2 was positively correlated with DANCR expression in ovarian cancer tissues. CONCLUSIONS: In this study, we revealed that DANCR could enhance the proliferation, migration and invasion capacities of ovarian cancer cells by upregulating IGF2. Our findings might offer a potential therapeutic choice for patients with ovarian cancer.

[The efficacy analysis of allogeneic hematopoietic stem cell transplantation in 48 leukemia patients with central nervous system leukemia].

Objective: To investigate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of leukemia patients also suffering from central nervous system leukemia (CNSL) . Methods: A total of 48 leukemia patients with central nervous system leukemia admitted to our hospital from May 2012 to December 2017 were retrospectively analyzed. Results: ① Including 22 cases of acute lymphocytic leukemia (ALL) , 21 cases of acute myeloid leukemia (AML) , and 5 cases of chronic myelogenous leukemia (CML) . Before transplantation, 19 patients achieved complete remission (CR) , and the rest 29 ones without remission. ②The conditioning regimen used TBI as the main protocol, and 6 patients were combined with whole brain and total spinal cord radiotherapy, 2 with Cyber knife treatment, and children with modified IDA combined with BUCY. ③All 48 patients were successfully transplanted, the median time for leukocyte engraftment was 14 (10-23) days, the median time for platelet transplant 16 (6-78) days. ④Bone marrow was evaluated 28 days after transplantation, all 48 patients reached CR, and DNA testing confirmed that they were all full donor chimerism. ⑤The median follow-up was 14 (2-69) months. Of them, 28 cases survived, 10 relapsed and the rest 3 had recurrence of CNSL after transplantation. One year after allo-HSCT, the overall survival (OS) of CR and non-CR groups were (77.3±10.0) % and (57.6±9.3) % (P=0.409) , respectively, the disease-free survival rates (DFS) were (71.2±11.0) % and (53.9±9.5) % (P=0.386) , respectively. The 1-year OS rates of ALL and AML groups after transplantation were (54.2±10.7) %, (80.1±8.9) %, respectively (P=0.200) , and DFS rates were (49.2±10.8) %, (75.0±9.7) % (P=0.190) , respectively. Conclusion: Allo-HSCT was safe and effective for leukemia patients with CNSL.

[Effects of CD34(+) selected stem cells for the treatment of poor graft function after allogeneic stem cell transplantation].

Objective: To evaluate the efficacy and safety of purified CD34(+) stem cell boost in the treatment of poor graft function (PGF) after allogeneic hematopoietic stem cell transplantation (HSCT) . Methods: 12 patients with poor graft function, reported in our hospital during January 2014 to March 2018, were retrospectively analyzed; The donors of 12 patients were HLA mismatched family members, and all treated with donor purified CD34(+) stem cell after G-CSF mobilization, calculating and statistical analyzing the purity of separation and the recovery rate of CD34(+) stem cells. The related complications and the recovery of blood cells after infusion were observed. Results: The purity of CD34(+) cells in the separation products was 92.0% (44.0%-97.0%) , and the recovery rate was 55.0% (45.0%-96.7%) . The median number of CD34(+) cells was 1.9 (0.9-4.4) ×10(6)/kg with CD3(+) cells as 0.6 (0.3-2.0) ×10(4)/kg. The median durations of white blood cells, platelet and red blood cells recoveries were 18 (14-39) , 29 (16-153) and 60 (9-124) days, respectively. All 12 patients didn't experience serious adverse reactions in the process of infusion, 10 patients achieved hematopoietic recovery, 1 case partial remission, 1 case no recovery, without occurrence of aggravated infection, graft versus host disease and other complications. Conclusion: The infusion of donor purified CD34(+) stem cell was a safe and effective method for PGF after allogeneic HSCT.

[Experiment research of natural killer cells amplification in vitro and the killing effect on ovarian cancer cells].

Objective: To amplify natural killer (NK) cells in vitro and explore its killing effect on ovarian cancer cells. Methods: (1) The separation of NK cells and identification. A total of 20 ml peripheral blood of one healthy volunteer was collected in Nov. 2015, Peking University People's Hospital. The peripheral blood mononuclear cells of normal volunteers were isolated, cultured in vitro and amplificated cultivation for 14 days with K562 cells transfected and expressing interleukin 21 (IL-21-K562) as nourish cells. The number and dynamic state of the growth cells were monitored during the cultured process. Cells were harvested and counted after 14 days cultured. The NK cells phenotypes were detected by flow cytometry. (2) The killing effect of NK cells on ovarian cancer cells: the ratio of effector cells (NK cells) and target cells (ovarian cancer cells and its control) was 50∶1, 20∶1, 10∶1, 5∶1 or 1∶1, NK cells killing effect on ovarian cancer cells was detected by the lactate dehydrogenase (LDH) release experiments. Results: (1) The results of NK cells establishment and phenotypic characterization: the cells were induced in vitro for 14 days by amplification culture. With the extension of incubation time, the number of NK cells increased constantly, from 2.0×10(7) on day 0 to 5.1×10(9) on day 14. Obvious amplification of the total number of cells were detected for 255 times. Living cells unstained by trypan blue eventually reached 95% above. Before and after the induction and amplification in vitro, the percentage of NK cells(CD(3)(-)CD(5)(6+)cells) in CD(3)- cells were 2.33% and 85.32%, respectively (P<0.01), which covered the whole lymphocytes 1.06% and 69.42%, respectively (P<0.01), which showed that NK was the main cell type in the amplificated lymphocytes. (2) The killing rate of NK cells on ovarian cancer cells in vitro: the results detected by LDH release experiments showed that NK cells could performed strong nonspecific killing effect on ovarian cancer cell lines SKOV3, HOC1A, 3AO and CAOV3, as well the normal ovarian cell line T29 and NK sensitive cell line K562, and the killing effect increased significantly along with the increase of effector cells and target cells ratio (P<0.01). When the ratio was 1∶1, the killing rate was 37% for K562, while the rate of killing of other cells was around 10% (P<0.05). When the effect-target ratio was 20∶1 and 50∶1, in addition to CAOV3 cells (more than 70%), NK cells had a kill rate of more than 80% for other ovarian cancer cells lines and their control cell K562 and T29 cells (P>0.05). Conclusion: NK cells could be established in vitro and have a good non-specific killing effect on ovarian cancer cells.

Synergism between RIZ1 gene therapy and paclitaxel in SiHa cervical cancer cells.

RIZ1 is a tumor suppressor gene. The purpose of the present study was to investigate the inhibitory effect of RIZ1 gene therapy on the growth of SiHa cervical cancer cells and its synergism with paclitaxel. The expression levels of RIZ1 were examined by real-time PCR and western blotting before and after transfection of RIZ1. The effects of paclitaxel or pcDNA3.1(+)-RIZ1 alone or in combination, on the proliferation of SiHa cells were evaluated by MTT method. The inhibitory effect on the proliferation of SiHa cells was more significant in the pcDNA3.1(+)-RIZ1 combined with paclitaxel group than in the pcDNA3.1(+)-RIZ1 or paclitaxel groups (P<0.05). The expression level of RIZ1 in SiHa cells increased after treatment with paclitaxel, which indicated a synergism between them. RIZ1 gene therapy combined with paclitaxel showed stronger cell inhibition than paclitaxel alone, which indicated a synergism between them.

Combined Anti-CD154/CTLA4Ig Costimulation Blockade-Based Therapy Induces Donor-Specific Tolerance to Vascularized Osteomyocutaneous Allografts.

Tolerance induction by means of costimulation blockade has been successfully applied in solid organ transplantation; however, its efficacy in vascularized composite allotransplantation, containing a vascularized bone marrow component and thus a constant source of donor-derived stem cells, remains poorly explored. In this study, osteomyocutaneous allografts (alloOMCs) from Balb/c (H2(d) ) mice were transplanted into C57BL/6 (H2(b) ) recipients. Immunosuppression consisted of 1 mg anti-CD154 on day 0, 0.5 mg CTLA4Ig on day 2 and rapamycin (RPM; 3 mg/kg per day from days 0-7, then every other day for 3 weeks). Long-term allograft survival, donor-specific tolerance and donor-recipient cell trafficking were evaluated. Treatment with costimulation blockade plus RPM resulted in long-term graft survival (>120 days) of alloOMC in 12 of 15 recipients compared with untreated controls (median survival time [MST] ≈10.2 ± 0.8 days), RPM alone (MST ≈33 ± 5.5 days) and costimulation blockade alone (MST ≈45.8 ± 7.1 days). Donor-specific hyporesponsiveness in recipients with viable grafts was demonstrated in vitro. Evidence of donor-specific tolerance was further assessed in vivo by secondary donor-specific skin graft survival and third-party graft rejection. A significant increase of Foxp3(+) regulatory T cells was evident in tolerant animals. Donor cells populated peripheral blood, thymus, and both donor and recipient bone marrow. Consequently, combined anti-CD154/CTLA4Ig costimulation blockade-based therapy induces donor-specific tolerance in a stringent murine alloOMC transplant model.

Stress analysis during jaw movement based on vivo computed tomography images from patients with temporomandibular disorders.

The purpose of this study is to develop three-dimensional (3D) finite element models of temporomandibular joints (TMJs) and to investigate stress distributions. To determine the causes of temporomandibular disorders (TMDs), the magnitude and location of the maximum stresses under physiological loading must be considered. Stress analysis TMD models were reconstructed from computed tomography (CT) data. Several studies have investigated finite element TMJ models, but few have used a bilateral mandible model that includes jaw closing and maximum opening. In this study, the authors defined an asymmetry index for the different stress values on each side joint; this index has not yet been investigated. According to clinical observation, one joint affects the other side joint during mastication. Three symptom-free volunteers and three symptomatic patients were selected as the control group (CG) and TMD group (TG), respectively. For the TG, data analysis indicated that the condyle was asymmetrical during jaw closing, while both the condyle and disc were slightly asymmetrical during jaw opening. The maximum stresses did not significantly differ between the CG and TG for either closing or opening of the jaw. The results of this study have a potential clinical benefit in terms of proving superior biomechanical behaviour.

Expression of beclin 1, an autophagy-related protein, in human cervical carcinoma and its clinical significance.

PURPOSE: To investigate the impact of beclin 1 on prognosis of cervical cancer, we determined the expression of beclin 1 in cervical cancer, cervical intraepithelial neoplasia (CIN) and normal cervical tissues. METHODS: A total of 122 cases of cervical cancer, 35 cases with CIN and 31 cases with uterine fibroids were collected at the Cancer Center of Sun Yat University to determine the expression of beclin 1. RESULTS: Beclin 1 positive rate in normal cervical tissues, CIN tissues and cervical cancers was 83.9%, 74.3% and 53.3%, respectively, and it was significantly different between the three groups (p < 0.01). Beclin 1 expression was negatively correlated with cervical cancer differentiation, lymph node metastasis, recurrence and death (p < 0.05). The negative expression is the risk factor affecting overall survival (p < 0.05) and progression-free survival (PFS) (p < 0.05). Multivariate analysis showed that beclin 1 negative expression was an independent risk factor of PFS time. CONCLUSIONS: Beclin 1 may play a role in the occurrence and development of cervical cancer. Beclin 1 positive expression in patients indicates a better prognosis.

The role of HE4 in ovarian cancer: inhibiting tumour cell proliferation and metastasis.

As a promising biomarker, human epididymis protein 4 (HE4) has been widely used for the early detection and differential diagnosis of ovarian cancer. This study evaluated the function of HE4 in the carcinogenesis and progression of ovarian cancer. An enzyme immunometric assay, used to detect HE4 in the serum of ovarian cancer patients, showed that the protein could discriminate between malignant and benign ovarian tumours with high specificity. An exogenous HE4 gene was transfected into ovarian cancer cell lines and an immortalized ovarian epithelial cell line. Compared with the controls, HE4 overexpression significantly promoted cell apoptosis and adhesion. Overexpression of HE4 also led to significant inhibition of cell proliferation, migration and invasiveness in vitro, as well as xenograft tumour formation in vivo. This is the first report to demonstrate the functional importance of HE4 in multiple cellular processes and indicates that HE4 may play a protective role in the progression of ovarian cancer.

DNA methylation of the RIZ1 tumor suppressor gene plays an important role in the tumorigenesis of cervical cancer.

OBJECTIVE: The retinoblastoma protein-interacting zinc finger gene RIZ1 is a tumor suppressor gene and a member of a nuclear histone/protein methyltransferase superfamily. The purpose of the present study was to examine the expression of RIZ1 and evaluate its carcinogenesis in cervical cancer. The relationship between DNA methylation and transcriptional silencing of RIZ1 was investigated in cervical cancer. METHODS: RIZ1 expression was examined in cervical cancer cell lines and cervical tissues (12 normal and 40 cancerous) by using RT polymerase chain reaction (PCR). Methylation status of the RIZ1 promoter was studied using methylation-specific PCR (MSP). RESULTS: RIZ1 expression is reduced or lost in cervical cancers, compared with normal cervical tissues (P <0.05). The current study results also showed that loss of RIZ1 is mediated by aberrant cytosine methylation of the RIZ1 promoter. 37.5% of carcinomas were methylated, while none of normal tissues were methylated. RIZ1 mRNA expression was significantly higher (P = 0.000) in unmethylated (0.3494 +/- 0.0466, mean +/-SD), compared with methylated tissues (0.1422 +/- 0.1073, mean +/-SD). Treatment with a DNA methyltransferase inhibitor led to reactivation of RIZ1 expression in cell lines that had negligible RIZ1 expression at baseline. CONCLUSIONS: Reduced expression of RIZ1 may play an important role in the pathogenesis and/or development of cervical cancer, and is considered to be caused in part by aberrant DNA methylation.

A point mutation in the N-terminal coiled-coil domain releases c-Fes tyrosine kinase activity and survival signaling in myeloid leukemia cells.

The c-fes locus encodes a 93-kDa non-receptor protein tyrosine kinase (Fes) that regulates the growth and differentiation of hematopoietic and vascular endothelial cells. Unique to Fes is a long N-terminal sequence with two regions of strong homology to coiled-coil oligomerization domains. We introduced leucine-to-proline substitutions into the coiled coils that were predicted to disrupt the coiled-coil structure. The resulting mutant proteins, together with wild-type Fes, were fused to green fluorescent protein and expressed in Rat-2 fibroblasts. We observed that a point mutation in the first coiled-coil domain (L145P) dramatically increased Fes tyrosine kinase and transforming activities in this cell type. In contrast, a similar point mutation in the second coiled-coil motif (L334P) was without effect. However, combining the L334P and L145P mutations reduced transforming and kinase activities by approximately 50% relative to the levels of activity produced with the L145P mutation alone. To study the effects of the coiled-coil mutations in a biologically relevant context, we expressed the mutant proteins in the granulocyte-macrophage colony-stimulating factor (GM-CSF)-dependent myeloid leukemia cell line TF-1. In this cellular context, the L145P mutation induced GM-CSF independence, cell attachment, and spreading. These effects correlated with a marked increase in L145P protein autophosphorylation relative to that of wild-type Fes. In contrast, the double coiled-coil mutant protein showed greatly reduced kinase and biological activities in TF-1 cells. These data are consistent with a role for the first coiled coil in the negative regulation of kinase activity and a requirement for the second coiled coil in either oligomerization or recruitment of signaling partners. Gel filtration experiments showed that the unique N-terminal region interconverts between monomeric and oligomeric forms. Single point mutations favored oligomerization, while the double point mutant protein eluted essentially as the monomer. These data provide new evidence for coiled-coil-mediated regulation of c-Fes tyrosine kinase activity and signaling, a mechanism unique among tyrosine kinases.

Azepanone-based inhibitors of human and rat cathepsin K.

The synthesis, in vitro activities, and pharmacokinetics of a series of azepanone-based inhibitors of the cysteine protease cathepsin K (EC 3.4.22.38) are described. These compounds show improved configurational stability of the C-4 diastereomeric center relative to the previously published five- and six-membered ring ketone-based inhibitor series. Studies in this series have led to the identification of 20, a potent, selective inhibitor of human cathepsin K (K(i) = 0.16 nM) as well as 24, a potent inhibitor of both human (K(i) = 0.0048 nM) and rat (K(i,app) = 4.8 nM) cathepsin K. Small-molecule X-ray crystallographic analysis of 20 established the C-4 S stereochemistry as being critical for potent inhibition and that unbound 20 adopted the expected equatorial conformation for the C-4 substituent. Molecular modeling studies predicted the higher energy axial orientation at C-4 of 20 when bound within the active site of cathepsin K, a feature subsequently confirmed by X-ray crystallography. Pharmacokinetic studies in the rat show 20 to be 42% orally bioavailable. Comparison of the transport of the cyclic and acyclic analogues through CaCo-2 cells suggests that oral bioavailability of the acyclic derivatives is limited by a P-glycoprotein-mediated efflux mechanism. It is concluded that the introduction of a conformational constraint has served the dual purpose of increasing inhibitor potency by locking in a bioactive conformation as well as locking out available conformations which may serve as substrates for enzyme systems that limit oral bioavailability.

Src homology 2 domain substitution modulates the kinase and transforming activities of the Fes protein-tyrosine kinase.

The c-fes proto-oncogene encodes a Mr 93,000 protein-tyrosine kinase (Fes) that is strongly expressed in myeloid cells and has been implicated in myelomonocytic differentiation. Fes autophosphorylation and transforming activity are highly restrained after ectopic expression in fibroblasts, indicating tight negative regulation of Fes kinase activity in vivo. Here we investigated the regulatory role of the Fes Src homology 2 (SH2) domain by producing a series of chimeric constructs in which the Fes SH2 domain was replaced with those of the transforming oncogenes v-Fps and v-Src or by the NH2-terminal SH2 domain of the Ras GTPase-activating protein. Wild-type and chimeric Fes proteins readily underwent tyrosine autophosphorylation in vitro and produced identical cyanogen bromide phosphopeptide cleavage patterns, indicating that the SH2 substitutions did not influence overall kinase activity or autophosphorylation site selection. However, metabolic labeling of Rat-2 fibroblasts expressing each construct showed that only the Fes/Src SH2 chimera was active in vivo. Consistent with this result, the Fes/Src SH2 domain chimera exhibited potent transforming activity in fibroblasts and enhanced differentiation-inducing activity in K-562 myeloid leukemia cells. In addition, the Fes/Src SH2 chimera exhibited constitutive localization to focal adhesions in Rat-2 fibroblasts and induced the attachment and spreading of TF-1 myeloid cells. These data demonstrate a central role for the SH2 domain in the regulation of Fes kinase activity and biological function in vivo.