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Objective: To explore the effect of warm-needle moxibustion (WNM) on the levels of T cell subgroups and serum inflammatory factors, intestinal microecological balance and postoperative adverse reactions in patients with colorectal cancer., Methods: Eighty-four patients who underwent elective radical resection of colorectal cancer were randomly and equally divided into control (medication) group (23 men and 19 women) and WNM group (24 men and 18 women). Patients of the control group received conventional medication treatment (such as postoperative anti-infection and fluid supplementation), and those of the WNM group received conventional medication plus WNM stimulation (the acupuncture needle handle warmed by ignited moxa stick) of bilateral Zusanli(ST36), Sanyinjiao(SP6), Yinlingquan(SP9), Shangjuxu(ST37), and Zhaohai(KI6). The acupuncture needles were retained for 45 minutes every time, starting on the first day after surgery, once a day for 15 days. The number of T cell subsets (CD3 + , CD4 + , CD8 + ) positive cells was counted under fluorescence microscope, and the contents of serum tumor necrosis factor α(TNF-α) and interleukin-6 (IL-6) were detected by using ELISA, and the level of C-reactive protein (CRP) was detected by using immunoturbidimetry. The levels (logarithm of colony-forming units per gram of wet fecal weight) of Bifidobacterium, Lactobacillus, Escherichia coli and Enterococcus were determined. The adverse reactions (leukocyte decline, nausea and vomiting, peripheral phlebitis, cold stimulation sensitivity) were recorded after surgery., Results: Before treatment, there were no significant differences between the two groups in the number of T cell subgroups, TNF-α and IL-6 contents, and intestinal flora numbers ( P >0.05). After the treatment, self-comparison showed that the numbers of CD3 + and CD4 + positive cells, the ratio of CD4 + /CD8 + and the intestinal Bifidobacterium and Lactobacillus levels in the WNM group were significantly increased ( P <0.05), whereas the number of CD8+positive cells, intestinal Escherichia coli and Enterococcus levels in the WNM group, and the levels of TNF-α, IL-6 and CRP in both groups were obviously decreased in comparison with their own pretreatment ( P <0.05), but no significant changes were found in the levels of CD3 + and CD4 + positive cells, CD4 + /CD8 + and intestinal Bifidobacterium, Lactobacillus, Escherichia coli and Enterococcus in the control group ( P >0.05). Comparison between two groups displayed that after the treatment, the numbers of CD3 + and CD4 + positive cells, the ratio of CD4 + /CD8 + , as well as the levels of Bifidobacterium and Lactobacillus were significantly higher in the WNM group than in the control group ( P <0.05), whereas the number of CD8 + positive cells， TNF-a, IL-6 and CRP, and the levels of Escherichia coli and Enterococcus were obviously lower in the WNM group than in the control group ( P <0.05). The incidence of adverse reactions including leukopenia, nausea and vomiting, peripheral phlebitis, and sensitivity to cold stimulation in the WNM group were markedly lower than those of the control group ( P <0.05)., Conclusion: WNM intervention can significantly improve the immune function, reduce the level of inflammatory factors, regulate the level of beneficial intestinal flora, and also reduce the incidence of postoperative adverse reactions in patients experiencing radical resection of colorectal cancer.

Objective: To evaluate the effect on postoperative complications of mixed hemorrhoids treated with electroacupuncture(EA) at Chengshan(BL 57)，Dachangshu(BL 25)and Erbai(EX-UE 2)., Methods: A total of 60 patients with mixed hemorrhoids were randomly divided into control and EA groups，30 cases in each group. In addition to basic treatment, EA was applied at BL 57, BL 25 and EX-UE 2 for 30 min (2 Hz/15 Hz，0.5－2 mA) each day for 3 days after surgery in the EA group, while the control group received basic treatment after surgery. Postoperative complications after treatment were evaluated each day., Results: Following surgery, anal pain, anal pendant expansion and hematochezia were observed in both the EA group and control group. On the first day after surgery, symptom improvement was not statistically significantly different between the two groups ( P >0.05)．On the 2 nd and 3 rd days after surgery, anal pain, anal pendant expansion and hematochezia were markedly improved in the EA group compared with those in the control group ( P <0.05)., Conclusions: Electroacupuncture can reduce anal pain, anal pendant expansion and hematochezia after surgery in patients with mixed hemorrhoids.

Aim: To improve an asialoglycoprotein receptor (ASGPR)-based enrichment method for detection of circulating tumor cells (CTCs) of hepatocellular carcinoma (HCC)., Methods: Peripheral blood samples were collected from healthy subjects, patients with HCC or various other cancers, and patients with hepatic lesions or hepatitis. CTCs were enriched from whole blood by extracting CD45-expressing leukocytes with monoclonal antibody coated-beads following density gradient centrifugation. The remaining cells were cytocentrifuged on polylysine-coated slides. Isolated cells were treated by triple immunofluorescence staining with CD45 antibody and a combination of antibodies against ASGPR and carbamoyl phosphate synthetase 1 (CPS1), used as liver-specific markers, and costained with DAPI. The cell slide was imaged and stained tumor cells that met preset criteria were counted. Recovery, sensitivity and specificity of the detection methods were determined and compared by spiking experiments with various types of cultured human tumor cell lines. Expression of ASGPR and CPS1 in cultured tumor cells and tumor tissue specimens was analyzed by flow cytometry and triple immunofluorescence staining, respectively., Results: CD45 depletion of leukocytes resulted in a significantly greater recovery of multiple amounts of spiked HCC cells than the ASGPR(+) selection (Ps < 0.05). The expression rates of either ASGPR or CPS1 were different in various liver cancer cell lines, ranging between 18% and 99% for ASGPR and between 9% and 98% for CPS1. In both human HCC tissues and liver cancer cell lines, there were a few HCC cells that did not stain positive for ASGPR or CPS1. The mixture of monoclonal antibodies against ASGPR and CPS1 identified more HCC cells than either antibody alone. However, these antibodies did not detect any tumor cells in blood samples spiked with the human breast cancer cell line MCF-7 and the human renal cancer cell line A498. ASGPR(+) or/and CPS1(+) CTCs were detected in 29/32 (91%) patients with HCC, but not in patients with any other kind of cancer or any of the other test subjects. Furthermore, the improved method detected a higher CTC count in all patients examined than did the previous method (P = 0.001), and consistently achieved 12%-21% higher sensitivity of CTC detection in all seven HCC patients with more than 40 CTCs., Conclusion: Negative depletion enrichment combined with identification using a mixture of antibodies against ASGPR and CPS1 improves sensitivity and specificity for detecting circulating HCC cells.

Objective: To observe the efficacy of comined administration of acupuncture and anesthetics for mixed hemorrhoids surgery and its postoperative complications., Methods: A total of 40 cases of mixed hemorrhoid patients were randomized into acupuncture-assisted anesthesia (AAA) group and narcotic drug group (control), with 20 cases in each. For patients of AAA group, electroacupuncture (EA, 2 Hz/100 Hz, 9 mA) was applied to Changqiang (GV 1) and Yaoshu (GV 2) for 20 min, and 1% lidocaine (5 mL) was injected into GV 1. For patients of control group, 20 mL and 10 mL of 1% lidocaine were injected into GV 2 and GV 1, separately. The patients' pain response, anus muscular relaxion state, and postoperative complications were observed to evaluate their clinical efficacy., Results: Of the 20 cases in AAA and control groups, 16 and 18 cases had a striking analgesic effect, 4 and 2 were effective, respectively. No significant difference was found between two groups in the analgesic efficacy for surgery (P>0.05). Postoperative urinary retention, pain severity I and II and the cases with intoxic response in AAA and control groups were 1 and 8, 16 and 6, 4 and 14, and 1 and 9, separately. The effects of AAA group were significantly superior to those of control group in lessening urinary retention and toxication, and in inducing analgesia (P<0.05)., Conclusion: Acupuncture combined with narcotic drugs is superior to simple anesthetics for mixed hemorrhoids surgery in reducing postoperative complications.

Aim: To examine the effect of troglitazone, a peroxisome proliferator-activated receptor gamma (PPAR gamma) ligand, on the proliferation and apoptosis of human liver cancer cells., Methods: Liver cancer cell line HepG2 was cultured and treated with troglitazone. Cell proliferation was detected by 3-(4-,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay; apoptosis was detected by flow cytometry and terminal deoxynucleotidyl transferase-mediated nick end labeling of DNA fragmentation sites (TUNEL) assay; and apoptosis-related protein was detected by immunocytochemistry and Western blotting., Results: Troglitazone inhibited growth and induced apoptosis of HepG2 cells in a dose-dependent manner, and induced activation of caspase-3 expression. Troglitazone not only drove apoptosis-inhibiting factor survivin to translocate incompletely from the nucleus to the cytoplasm, but also inhibited expression of survivin, while it did not affect expression of apoptosis-promoting factor Bax., Conclusion: PPAR gamma ligands inhibit growth and induce apoptosis of liver cancer cells, and may have applications for the prevention and treatment of liver cancer.

Objective: To investigate the specific cytotoxity of tumor infiltrating lymphocytes (TIL) transfected with chimeric T cell receptor (CTCR) on cells which express KDR., Methods: A recombinant retroviral plasmid (pMSCVneo-Vhgamma) was constructed by cloning VEGF121-hinger-FcRgamma (Vhgamma) into retroviral vector pMSCVneo. After packaging by PT67, the virus with high titer was used to infect TIL isolated from liver cancer tissues, and then MSCVneo-Vhgamma-TIL was generated. TIL infected with MSCVneo was used as a control. The cytotoxicty of the transgenic TIL on NIH3T3 and HepG2 expressing no KDR and on ECV304 and A375 expressing KDR was detected with MTT colorimetric assay., Results: The sequences of VEGF121 and hinger-FcRgamma were different from those reported, but the deduced amino sequences were identical to the reported ones. The cytotoxity of TIL infected with MSCVneo on target cell was similar to that of the control TIL; both only had mild cytotoxity on cancer cell line. No significant cytotoxity was found in TIL infected with MSCVneo-cTCR on NIH3T3, but its cytotoxity on ECV304 was significant. The cytotoxity on HepG2 was similar to that of MSCVneo-TIL and uninfected TIL, but cytotoxity on A375 was significantly higher., Conclusion: Chimeric T cell receptor permanently grafts TIL cell with predefined new specificity. TIL expressing Vhgamma can selectively recognize and kill vascular endothelial cell and tumor cells which express VEGF receptor KDR.

Objective: To investigate the relationship between serum alpha-fetoprotein (AFP) variant levels in patients with hepatocellular carcinoma (HCC) and cancer cells disseminating through blood., Methods: Serum AFP variant levels were measured by crossed immunoaffino-electrophoresis in the presence of lectin before initial surgical treatment in HCC patients. Circulating tumor cells were simultaneously detected in pre-operative blood samples using reverse transcription-polymerase chain reaction (RT-PCR) for AFP mRNA., Results: Forty-six HCC patients with serum AFP positive were studied. Serum AFP variant level > or 20% was showed in 37 patients, among whom there were 22 (59.5%) showing AFP mRNA positive. In contrast, the positive AFP mRNA expression was only observed in 2 out of 9 patients (22.2%) with AFP variant level<20% (x(2)=4.02, P<0.05)., Conclusion: In hepatocellular carcinoma patients, increased AFP variant levels are associated with a haematogenous spread of tumor cells.