PEG-rhG-CSF for prophylaxis of neutropenia after chemotherapy in patients with non-small cell lung cancer: A multicenter, prospective, randomized study.

BACKGROUND: To evaluate the efficacy and safety of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in preventing neutropenia during multiple cycles of chemotherapy in patients with non-small cell lung cancer (NSCLC). METHOD: In a multicenter, prospective, randomized trial, patients with NSCLC were randomly assigned in a 2:1 ratio to treatment group (PEG-rhG-CSF as primary prophylactic therapy) or control group. Patients in the control group were administered rhG-CSF when white blood cell count was <2.0 × 109 /L or absolute neutrophil count <1.0 × 109 /L. The primary endpoint was the incidence of grade 3/4 neutropenia. Secondary endpoints included the incidence and duration of grade 3/4 neutropenia in each cycle, the incidence of febrile neutropenia (FN), delay rate of chemotherapy, prolonged time of chemotherapy, and safety. RESULTS: Between January 2019 and July 2021, 130 patients were enrolled (treatment group: n = 87, control group: n = 43). The incidence of grade 3/4 neutropenia in the treatment group was significantly lower than that in the control group (1.15% vs. 11.63%, p < 0.05). The mean duration of grade 3/4 neutropenia for the treatment and control group was 2.00 and 3.75 days, respectively. There were no statistical differences in the incidence of FN, delay rate of chemotherapy, prolonged time of chemotherapy, and antibiotic use between the two groups (all p > 0.05). Adverse events were reported in 47.13% of patients in the treatment group and 48.84% patients in the control group. CONCLUSIONS: Primary prophylactic treatment with PEG-rhG-CSF could reduce the incidence of neutropenia in patients with NSCLC during multiple cycles of chemotherapy, with acceptable safety and tolerability.

Expression and significance of transforming growth factor-ß receptor type II and DPC4/Smad4 in non-small cell lung cancer.

The aim of the present study was to investigate the expression levels of transforming growth factor-ß (TGF-ß) receptor type II (TßRII) and DPC4/Smad4 in the TGF-ß signaling pathway and the importance of these expression levels in non-small cell lung cancer (NSCLC). The mRNA and protein expression levels of TßRII and DPC4/Smad4 were detected by reverse transcription-quantitative polymerase chain reaction and western blotting, respectively, in NSCLC and control nonlesional lung tissues of 60 patients. The protein expression levels of DPC4/Smad4 were detected by immunohistochemistry in paraffin-embedded samples of NSCLC. In addition, the correlations among the expression levels of TßRII and DPC4/Smad4 and their association with the clinical and pathological features of NSCLC were analyzed. The expression levels of TßRII and DPC4/Smad4 in NSCLC tissues were significantly lower when compared with the control nonlesional lung tissues (P<0.05). In addition, the expression of TßRII and DPC4/Smad4 in poorly-differentiated NSCLC tissues was significantly lower compared with moderately- or well-differentiated NSCLC tissues (P<0.05). The expression levels of TßRII and DPC4/Smad4 were significantly lower in NSCLC tissues with metastatic lymph nodes compared with tissue without metastatic lymph nodes (P<0.05). Thus, the expression levels were demonstrated to significantly correlate with the clinical and pathological stages, and subsequently were shown to be associated with the occurrence and progression of NSCLC. In conclusion, TßRII and DPC4/Smad4 may play an important role in the tumorigenesis, differentiation and progression of NSCLC via the TGF-ß signaling pathway.

Laribacter hongkongensis isolated from a patient with community-acquired gastroenteritis in Hangzhou City.

We describe the isolation of Laribacter hongkongensis in Hangzhou City, People's Republic of China. One strain of bacterium, named LHHZ242, had many of the same phenotypic and genotypic characteristics as Laribacter hongkongensis described in previous publications. This discovery proves that Laribacter hongkongensis is also associated with community-acquired gastroenteritis outside Hong Kong.

[The effect of siRNA-Her2/neu on the drug sensitivity of Her2/neu-overexpressing lung adenocarcinoma cell line].

OBJECTIVE: To explore the effect of synthesized siRNA targeting Her2/neu oncogene on the drug sensitivity of Her2/neu-overexpressing lung adenocarcinoma cell line. METHODS: The experiments consisted of four groups including an untreated control group, an empty vector group, an unrelated siRNA group, and a Her2/neu siRNA group. Every experiment was repeated five times. Lung adenocarcinoma cell line Calu-3 was transfected with siRNAs formulated LipofectAMINE 2000, and Her2/neu protein and P-gp of each group were determined by flow cytometry (FCM). The chemosensitivity of transfected cells to cisplatin (DDP) was measured by MTT. Cell apoptosis detection kit (Annexin V method) was used to examine the drug induced apoptosis rate. RESULTS: The Her2/neu protein and P-gp positive expression rate in the Her2/neu siRNA group, the untreated control group, the empty vector group and the unrelated siRNA group were [(25.04 +/- 1.56)%, (4.24 +/- 1.01)%], [(98.24 +/- 2.23)%, (5.11 +/- 2.98)%], [(95.67 +/- 1.98)%, (6.98 +/- 2.47)%] or [(94.79 +/- 0.87)%, (5.59 +/- 3.66)%], respectively. Introduction of the sequence specific siRNA into Her2/neu positive Calu-3 cells in vitro greatly reduced the cell surface expression of the Her2/neu protein, but had no effect on P-gp level. Consequently the inhibitory rate of DDP in combination with siRNA targeting Her2/neu was (67.1 +/- 2.3)%, but it was (48.1 +/- 3.5)%, (46.3 +/- 5.9)% and (50.2 +/- 2.9)% in the untreated control, the empty vector and the unrelated siRNA groups, respectively. There was a significant difference between Her2/neu siRNA group and other three groups (P < 0.01). The FCM results showed the apoptosis rate of DDP combined with siRNA-Her2/neu was elevated as compared with the unrelated siRNA group, the empty vector group or the untreated control group. CONCLUSION: Sequence specific siRNA-Her2/neu was capable of enhancing the chemosensitivity of Calu-3 cells to cisplatin.

[Effect of RNAi-mediated gene silencing of C-erbB-2 on proliferation of lung adenocarcinoma cell line calu-3].

BACKGROUND & OBJECTIVE: C-erbB-2 gene is amplified or overexpressed in breast cancer, ovarian cancer, and lung cancer, and is related with enhanced malignancy and metastatic ability, intrinsic chemoresistance, and poor prognosis of tumors. RNA interfering (RNAi), a new genetic technique, can efficiently and specifically suppress gene expression. This study was to investigate the effect of small interfering RNA (siRNA)-mediated gene silencing of C-erbB-2 on proliferation of human lung adenocarcinoma cell line calu-3. METHODS: C-erbB-2 siRNA was transfected into calu-3 cells; cell morphology was observed under light microscope. The mRNA and protein levels of C-erbB-2 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry (FCM). The proliferation of calu-3 cells was assessed by MTT assay. Cell cycle and apoptosis were analyzed by FCM. RESULTS: C-erbB-2 siRNA down-regulated the mRNA and protein levels of C-erbB-2 in calu-3 cells; 48 h after transfection of C-erbB-2 siRNA, the protein level of C-erbB-2 was markedly decreased. The positive rate of C-erbB-2 was significantly lower in C-erbB-2 siRNA group than in untransfected group, empty vector group, and nonspecific siRNA group [(25.04+/-1.56)% vs. (98.24+/-2.23)%, (95.67+/-1.98)%, and (94.79+/-0.87)%, P < 0.01]. C-erbB-2 siRNA inhibited proliferation of calu-3 cells: G(0)/G(1) phase proportion of C-erbB-2 siRNA group was significantly higher than that of untransfected group [(56.6+/-3.6)% vs. (45.5+/-3.2)%, P < 0.01]. C-erbB-2 siRNA also enhanced cell apoptosis. CONCLUSION: Specific siRNA targeting C-erbB-2 can effectively inhibit C-erbB-2 expression and proliferation of calu-3 cells.

[Effects of Her-2/neu siRNA-mediated gene silencing on cell cycle and apoptosis of lung adenocarcinoma cells].

OBJECTIVE: To investigate the effect of synthesized Her-2/neu specific siRNA on the cell cycle and apoptosis of Her-2/neu upregulating human lung adenocarcinoma cells. METHODS: Human lung cancer cells of the line calu-3 were cultured and divided into 4 groups: untreated control group, blank vector group transfected with blank vector, non-specific siRNA group transfected with unrelated siRNA, and Her2/neu siRNA group transfected with Her2/neu siRNA. RT-PCR was used to examine the Her-2/neu and cyclin D(1) mRNA expression. Flow cytometry was used to examine the Her-2/neu protein expression and cell cycle. The apoptosis rate was analyzed by using annexin V-FITC kit. The vascular endothelial growth factor (VEGF) level in the culture supernatant was detected by ELISA. RESULTS: Twenty-four hours after transfection, the expressions of Her-2/neu mRNA and cyclin D1 mRNA in the Her2/neu siRNA group decreased remarkably, both significantly lower than those in the other 3 groups. Forty-eight hours after transfection, the expression rate of Her-2/neu protein was 25.0% +/- 1.6% in the calu-3 cells transfected with Her-2/neu siRNA, significantly lower than in the control group, blank vector group, and non-specific siRNA group (98.2% +/- 2.2%, 95.7% +/- 2.0%, and 94.8% +/- 1.6% respectively, all P < 0.01); the proportion of the cells in G(0)/G(1) stage increased and those in the S stage decreased in the celu-3 cells transfected with Her-2/neu siRNA, and the proportions of the cells in G(0)/G(1) stage and Stage did not significantly change (F = 6.1, P < 0.01); the apoptotic rate of the Her2/neu siRNA group was 25.1% +/- 1.2%, significantly higher than those of the other 3 groups (4.8% +/- 0.5%, 8.6% +/- 0.9%, and 10.3% +/- 0.3% respectively, all P < 0.01); the caspase-3 activity ratio of the Her2/neu siRNA group was 134.6% +/- 4.5%, significantly higher than those in the blank vector group and non-specific siRNA group (105.0% +/- 2.5% and 112.0% +/- 2.8% respectively, both P < 0.01), and the VEGF level in the supernatant of the Her2/neu siRNA group was 176 pg/ml +/- 6 pg/ml, significantly lower than those of the control group, blank vector group, and non-specific siRNA group (476 pg/ml +/- 13 pg/ml, 426 pg/ml +/- 9 pg/ml, and 406 pg/ml +/- 9 pg/ml respectively, all P < 0.01). CONCLUSION: Chemically synthesized Specific Her-2/neu targeting siRNA effectively inhibits Her-2/neu expression and leads to the decline of cyclin D(1) and VEGF levels and activation of caspase-3 pathway thus arresting the cell cycle at G(0)/G(1) stage and enhancing cell apoptosis.