Suitable T stage for cryosurgery to spare the anus in patients with low rectal cancer.

Sphincter preserving therapy is a key research focus for treating low rectal cancer; however, the role of cryotherapy in this process has seldom been reported in the literature. Therefore, we conducted a comprehensive report on the role of cryoablation in sphincter preservation and explored its effect in rectal cancers. An observational study used longitudinal observation and follow-up. Participants were screened from patients whose medical records showed cryotherapy intervention for low rectal cancers from January 2016 to December 2020, with more than 2 years of follow-up. The primary endpoint was progress-free survival, and the secondary outcomes were mainly related to sphincter preservation rate and complications. Thirty-five patients were enrolled in this study, all of whom had their sphincters preserved. Until June 2022, 35 cases achieved long-term progression-free survival (41.77 ± 15.58), with no recurrence observed in 88.57% (31/35) of all patients at follow-up. Cryotherapy showed no significant differences in progress-free survival between sexes (p > 0.05). Cox regression was used to analyze the factors affecting local recurrence, with sex, T stage, size, and cryo-time taken as covariates. The results showed that T stage was a risk factor for local recurrence (p = 0.01, odds ratio: 16.27, 95% confidence interval: 8.20,145.75). Analysis of the T stage according to different subgroups showed that T3 stage was an independent risk factor (p = 0.002). We observed seven cases of complications, which were classified into grades I-II. In patients with low rectal cancers, cryotherapy can safely and effectively preserve the anus and avoid low anterior resection syndrome. Cryoablation has a better curative effect on radical treatment, especially for tumors in the T0-2 N0M0 stage.

A novel treatment approach for a patient with advanced low rectal cancer: A case report and literature review.

RATIONALE: In the treatment of low rectal cancer (LRC), preserving the anal sphincter is increasingly attracting the attention of colorectal surgeons. Many patients refused to perform a colostomy. Here, we report a case of LRC in a middle-aged woman and the clinical implications of the symptom, the treatment process of LRC, and the complications. PATIENT CONCERNS: A 46-year-old woman visited our department with a tumor found on her physical examination because of hemafecia. Then she refused to perform abdominoperineal resection. DIAGNOSIS: The patient first completed a colonoscopy and then underwent a rectal biopsy. The tumor was diagnosed as a rectal adenocarcinoma after pathological evaluation. Then it was staged by magnetic resonance imaging and enhanced computed X-ray tomography. INTERVENTIONS: The treatment consisted of chemoradiotherapy followed by cryoablation. OUTCOMES: The patient achieved a good oncological outcome and preserved the sphincter successfully. The post-cryoablation course of the patient was uneventful and he remained healthy at the 1-year follow-up. LESSONS: The preservation of anal sphincters has attracted more and more attention from colorectal surgeons. From the patient's perspective, the preservation of the anal sphincter was a key part of her treatment. We should try to meet the wishes of patients on the basis of curing the disease.

Cryotherapy for low rectal and anal cancer: recommendation and indications.

Low rectal cancer is a common gastrointestinal malignancy. Organ preservation in the treatment of low rectal cancer is a challenge. By combining surgical resection with freezing-a complementary treatment for low rectal cancer-the anus can be preserved in some patients. However, we lack unified standards for colorectal cancer cryotherapy. Our hospital has been treating patients with cryotherapy since 1976. In our department, the indications for and contraindications to low rectal and anal cancer treatment are well established. In this paper, we summarize the indications for and contraindications to cryotherapy for colorectal cancer by reviewing the literature, drawing on our experience, and considering current imaging and histological techniques. Our aim is to facilitate clinical discussion and promote appropriate treatment.

Development of a method for comprehensive and quantitative analysis of armillarisin succinate ester in its medicinal preparations by liquid chromatography-ion trap mass spectrometry.

A rapid and sensitive method for the identification and quantification of armillarisin succinate ester (ASE), which is a patent drug, is described. The method used liquid chromatography-ion trap mass spectrometry (LC-IT-MS/MS). The ASE standard solution was directly infused into IT-MS for collecting MS(n) spectra. The major fragment ions of ASE were confirmed by MS(n) at m/z 333, 233, 202, 189 and 163 in negative ion mode, and m/z 335, 217, 189,175 and 161 in positive mode, respectively. The possible main fragment ions cleavage pathways were studied between in positive-ion mode and in negative-ion mode. Quantification was performed using the transitions m/z 333 --> 233 in negative mode. The method is reliable and reproducible, and the detection limit is 2 ng/mL. The method was validated in the concentration range of 1.0 - 50 microg/mL, intra- and inter-day precision ranged from 1.98% to 4.06%, and the accuracy was 97.5-106.2%. The mean recovery of ASE was 97.2-105.3% with RSD less than 4.35%. A LC-IT-MS/MS has successfully applied to determine the ASE in its medicinal preparations.

Expression of Oct4 in HCC and modulation to wnt/ß-catenin and TGF-ß signal pathways.

Hepatocellular carcinoma (HCC) is considered as a disease of dysfunction of the stem cells. Studies on stem cells have demonstrated that Oct4 plays a pivotal role in embryo regulation. In order to understand the role of Oct4 in HCC and the relationship among Oct4 and wnt/ß-catenin and TGF-ß signal pathways, we have detected the expression of Oct4, Nanog, Sox2, STAT3 as well as the genes in wnt/ß-catenin, and TGF-ß families in HCC cell lines and in tumor specimens from HCC patients. The authors found that Oct4 was expressed in all of the four HCC cell lines and the tumor specimens from HCC patients. Some other genes were also expressed in them with different level including Nanog, Sox2, STAT3 and TCF3, wnt10b, ß-catenin, ELF, Smad3 and Smad4. The ability of the clone formation and migration of the HepG2 decreased after Oct4 was knockdowned. Silencing of Oct4 and TCF3 in HCC cell line HepG2 revealed that there were complicated relationships among Oct4, wnt/ß-catenin family and TGF-ß family genes. Knockdowning Oct4 reduced the expression of TGF-ß family genes ELF, Smad3, Smad4 and wnt/ß-catenin family genes, wnt10b, and ß-catenin but increased TCF3. In reverse, knockdowning TCF3 led to the increased expression of Oct4 and TGF-ß family genes. In conclusion, the expression of Oct4 in HCC may play an important role as in stem cell. Because Oct4 improves not only the function of wnt/ß-catenin, but also the TGF-ß signal pathways, the significance of its expression in HCC might be more complicated than we evinced before.

Identification and quantification of the volatile constituents in Cnidium monnieri using supercritical fluid extraction followed by GC-MS.

The volatile components of Cnidium monnieri were obtained by supercritical fluid extraction (SFE) and analyzed by GC-MS (identification and determination of metabolites). The compounds were identified according to their retention times and mass spectra. The effects of different parameters, such as extraction pressure, temperature, dynamic extraction time, flow rate of CO(2), on the SFE of C. monnieri extracts were investigated. A total of 14 compounds of SFE extracts were identified. Osthole (69.52%), bornyl acetate (10.03%), alpha-pinene (4.71%), and imperatorin (2.42%) were the major compounds identified in C. monnieri SFE extracts. The quantitation of osthole and imperatorin were then accomplished. The linear calibration ranges were all 5-1000 microg/mL for osthole and imperatorin by GC-MS analysis. The recovery of osthole and imperatorin were in the range 96.5-101.8%. The LODs for osthole and imperatorin were 1.0 and 0.6 microg/mL, respectively.

[Analysis on the expression of the stem cells related genes in hepatocellular carcinoma cell lines].

OBJECTIVE: To investigate the expression of Oct4, Sox2, Nanog, SMO, beta-Catenin and Wnt5b mRNA in four hepatocellular carcinoma cell lines of SMMC-7721, Bel-7402, HepG2, MHCC-97 and normal hepatocellular cell line of L02, and to compare the response of these cell lines to all-trans retinoic acid. METHODS: RT-PCR was used to detect expression of Oct4, Sox2, Nanog, SMO, beta-Catenin and Wnt5b mRNA in four hepatocellular carcinoma cell lines and normal hepatocellular cell line. Real time-PCR was used to quantify the expression of the genes. RESULTS: There are different levels of expression of the stem cell-related gene in hepatocellular carcinoma cell lines and control cell line (P less than 0.05). There are significant differences in HepG2 and L-02 for the response to all-trans retinoic acid (P less than 0.05). CONCLUSIONS: The stem cell-related genes are differentially expressed in different hepatocellular carcinoma cell lines.

Anticancer drugs are synergistic with freezing in induction of apoptosis in HCC cells.

Cryotherapy has been shown to be an important therapeutic alternative to surgery in the treatment of hepatocellular carcinoma (HCC). Here, the influence of cryo-chemotherapy on HCC was examined in vitro using the human HCC cell line Bel-7402, a drug-resistant HCC cell line originating from Bel-7402 cells (Bel-7402/R), as well as two control cell lines, the HCC cell line SMMC-7721 and a colorectal tumor cell line HIC-251. Cells were treated with either exposure to different freezing temperatures (ranging from -15 to -80 degrees C for 20 min), exposure to sub-lethal concentrations of anticancer chemotherapy drugs or a combination of cryotherapy and chemotherapy. Cell viability and apoptosis under each condition were investigated. We found that the combined treatment resulted in increases in both cell death and apoptosis compared to either treatment alone. The increased level of apoptosis observed in Bel-7402 cells after cryo-chemotherapy was inhibited in the presence of caspase inhibitors. Furthermore, Bax expression was increased 2- to 3-fold in cells exposed to the combination treatment compared with cells treated by freezing or drugs alone. In contrast, Bcl-2 levels remained constant. Although Bel-7402/R cells originated from the Bel-7402 cell line, they were more sensitive to the freezing procedure than the parental cell line. The level of Bax expression in Bel-7402/R cells was also higher than that observed in the parental cell line. In addition, we found that Bel-7402/R cells had lower levels of survivin mRNA than the parental Bel-7402 cells, in both untreated and treated cells. In conclusion, our data show that in HCC cells, apoptosis induced by cryotherapy can be synergistically enhanced using anticancer drugs.

Stability of tramadol with three 5-HT3 receptor antagonists in polyolefin bags for patient-controlled delivery systems.

BACKGROUND: Mixing 5-hydroxytryptamine-3 (5-HT3) receptor antagonists with patient-controlled analgesia (PCA) solutions of tramadol has been shown to decrease the incidence of nausea and vomiting associated with the use of tramadol PCA for postoperative pain. However, such mixtures are not commercially available, and the stability of the drug combinations has not been duly studied. The study aimed to evaluate the stability of tramadol with three 5-HT3 receptor antagonists in 0.9% sodium chloride injection for PCA administration. MATERIALS AND METHODS: Test samples were prepared by adding 1,000 mg tramadol hydrochloride, 8 mg ondansetron hydrochloride, and 6 mg granisetron hydrochloride or 5 mg tropisetron hydrochloride to 100 mL of 0.9% sodium chloride injection in polyolefin bags. The samples were prepared in triplicates, stored at either 25°C or 4°C for 14 days, and assessed using the following compatibility parameters: precipitation, cloudiness, discoloration, and pH. Chemical stability was also determined using a validated high-pressure liquid chromatography method. RESULTS: All of the mixtures were clear and colorless throughout the initial observation period. No change in the concentration of tramadol hydrochloride occurred with any of the 5-HT3 receptor antagonists during the 14 days. Similarly, little or no loss of the 5-HT3 receptor antagonists occurred over the 14-day period. CONCLUSION: Our results suggest that mixtures of tramadol hydrochloride, ondansetron hydrochloride, granisetron hydrochloride, or tropisetron hydrochloride in 0.9% sodium chloride injection were physically and chemically stable for 14 days when stored in polyolefin bags at both 4°C and 25°C.

CIK cells from patients with HCC possess strong cytotoxicity to multidrug-resistant cell line Bel-7402/R.

AIM: To investigate the cytotoxicity of the cytokine-induced killer (CIK) cells from the post-operation patients with primary hepatocellular carcinoma (HCC) to multidrug-resistant (MDR) cell of HCC both in vitro and in vivo. METHODS: A drug-resistant cell line was established by culturing human HCC cell line Bel-7402 in complete RPMI 1640 medium with increasing concentrations of adriamycin from 10 to 2,000 nmol/L. CIK cells were obtained by inducing the peripheral blood mononuclear cells with rhIFN-gamma, monoclonal anti-CD3 antibody, rhIL-1alpha as well as rhIL-2, which were added into the culture. To detect the cytotoxicity of the CIK cells from HCC patients, the Bel-7402/R was taken as target (T) cells and CIK cells as effect (E) cells. Cytotoxic test was performed and measured by MTT. As to in vivo test, CIK cells were transfused into patients with HCC. The tumor specimens of the patients were obtained and immunohistochemistry was carried out to detect CD3, CD45, CD45RO as well as CD68. RESULTS: A MDR 1 HCC cell line Bel-7402/R was established. Its MDR1 mRNA overexpressed which was shown by RT-PCR; the P-glycoprotein expression increased from 1.32% of parent cells to 54%. CIK cells expanded vigorously by more than 70-fold and the CD3+CD56+ increased by more than 600-fold after 3-wk incubation on average. The cytotoxicity of CIK from HCC patients to Bel-7402/R was about 50% and to L-02 below 10% (t = 8.87, P<0.01), the same as that of CIK from normal individuals. Each of the 17 patients received 1-5 x 10(10) of CIK cell transfusion. No side effects were observed. After CIK treatment, the tumor tissue nodules formed and a large amount of lymphocytes infiltrated in the liver cancer tissue and CD3, CD45, CD45RO, and CD68 increased greatly which was shown by immunohistochemistry. CONCLUSION: A stable MDR1 HCC cell line has been established which could recover from liquid nitrogen and CIK from HCC patients has strong cytotoxicity to MDR HCC cell. CIK adoptive immunotherapy is safe and has no side effects. Receivers improved their immunity to tumor evidently. CIK treatment may be a better choice for HCC patients after operation to prevent the recurrence, especially when tumors have developed drug resistance.

Increased expression of IL-6 mRNA in hepatocellular carcinoma cell lines correlates with biological characteristics.

BACKGROUND AND AIMS: IL-6 has been implicated in both virus-associated and diethylnitrosamine-induced hepatocellular carcinomas (HCCs). Generally it is produced by immune cells such as Kupffer cells in the liver. To understand mechanisms by which IL-6 might participate in the genesis of HCCs, the production of IL-6 by cell lines under different conditions was examined to determine inducing factors. METHODS: Expression of IL-6 mRNA in both hepatoma cell lines and a normal liver cell line L-02 was measured by quantitative RT-PCR. Biological molecules including liposome, dsRNA and cell debris were used to stimulate IL-6 mRNA expression in HepG2 cells and inhibition was effected by RNAi. Proliferation was assessed by MTT and clone formation and migration was determined by scratch assay. RESULTS: All of the HCC cell lines observed expressed IL-6 mRNA, including HepG2, Bel-7402(7402), MHCC-97H and SMMC-7721.Normal liver cell line L-02 also expressed IL-6 mRNA. SiRNA to IL-6 specifically knockdowned IL-6 mRNA expression in HepG2, and liposome, dsRNA and cell debris increased it. Both proliferation and migration of HepG2 cells were related to the level of IL-6 HepG2 expressed. CONCLUSION: Both normal liver cell line and HCC cell lines can produce IL-6 so that Kupffer cells are noit the only source of the cytokine in the liver well as other immune cells. That the fact that HCC cells reacted to stimulation of biological molecules such as liposome, dsRNA or cell debris with increasing production of IL-6 indicates that the cytokine might play an important role not only in the period of tumor initiation but progression and recurrence as well.